Aim To investigate the nephroprotective activity of berberine in diabetic nephropathy （DN） mice. Methods Diabetic nephropathy was induced by intraperitoneal injection with 55 mg · kg^-1 streptozotocin（STZ） , ...Aim To investigate the nephroprotective activity of berberine in diabetic nephropathy （DN） mice. Methods Diabetic nephropathy was induced by intraperitoneal injection with 55 mg · kg^-1 streptozotocin（STZ） , Berberine was administered at daily doses of 50, 100 and 200 mg· kg^-1 by gavage for 8 weeks. To detect serum creatinine and blood urea nitrogen （BUN） levels, blood were collected after the last dose of berberine, renal cortex was separated on ice after heart peffusion by precooled normal saline. The specimen was stored in -80℃ for the next experiments, and some of the kidney tissue were immobilized by 4% paraformaldehyde solution and 3% glut- araldehyde solution for the preparation of paraffin tissue slides and electron microscope biopsy respectively. After that, PAS staining and electron microscope were used to observe the glomerular morphology changes; ELISA was used to measure proinflammatory chemokines and cytokines levels in renal cortex. Real time RT-PCR was taken to detect the level of nucleotide binding oligomerzation domain 2 （NOD2） mRNA, Western blot was used to test the ex- pression of NOD2 and autophagy marker light chain 3 （LC3） in renal cortex. Results Histopathological changes and the increase in serum creatinine and BUN in DN mice were significantly ameliorated by berberine in a dose-de- pendent manner. Additionally, The expression of tumor necrosis factor-or （TNF-α）, interleukin-6 （IL-6） and in- tercellular adhesion molecule-1 （ICAM-1） was markedly suppressed by berberine, indicating the inhibition of in- flammatory response. Treatment of DN mice with berberine also significantly reduced the expression of NOD2 and LC3 in the kidneys. Conclusion The current study showed the nephroprotective activity of berberine in DN mice could be attributed to the inhibition of inflammation and展开更多
OBJECTIVE To investigate the effect of Urena lobataleaves extract on the inhibition of nephropathy diabetic complication.METHODS This study uses control group post test only with male Sprague dawley rats.Diabetic rats...OBJECTIVE To investigate the effect of Urena lobataleaves extract on the inhibition of nephropathy diabetic complication.METHODS This study uses control group post test only with male Sprague dawley rats.Diabetic rats was induced by high fructose diet(HFD)and single dose streptozotocin 25mg·kg-1 bw intra peritoneal.The rat was administrated orally with water extract of U.lobataleaves in concentrations of 250,500 and 1000mg·kg-1 bw for 4 weeks.After scarifying,kidney organ were collected and then superoxyde dismutase(SOD)kidney level,malondialdehyda(MDA)and tumor necrosis factor-alpha(TNF-α)were examined.The data was analyzed using ANOVA test continued with LSD test(P<0.05).RESULTS The oral administration of U.lobataleaves extract 250,500 and 1000mg·kg-1 bw were able to increase SOD kidney level about 30%,60% and 90% respectively compared to diabetic group(P<0.05),while the MDA kidney level was decreased by 30%,60% and 70%(P<0.05)respectively.The supplementation of water extract from U.lobatain dose of 250,500 and 1000mg·kg-1 bw were also decrease TNF-αkidney level approximately 30%,40% and 60% compared to control group(P<0.05).In diabetic groups,SOD kidney level was decreased compared to normal group(P<0.05)while the MDA and TNF-αwere increased(P<0.05).CONCLUSION U.lobata leaves extract could inhibit nephropathy diabetic complication by increasing of SOD kidney level,decreasing of MDA kidney level,and TNF-α.This effect may be related to active compounds that act as an antioxidant and anti-inflammatory in U.lobata extract.展开更多
OBJECTIVE Diabetic nephropathy(DN)has been one of the most common complications of diabetes and the leading cause of end-stage renal disease.Glomerular hyperfiltrationis central in earlystage of DN and leads to the pr...OBJECTIVE Diabetic nephropathy(DN)has been one of the most common complications of diabetes and the leading cause of end-stage renal disease.Glomerular hyperfiltrationis central in earlystage of DN and leads to the progression of renal architectonic and functional abnormalities.Salvianolic acid A(SalA)has been proved to protect diabetic complications such as hepatic fibrosis and neuropathy.The present study was designed to investigate the effects of SalA on glomerular endothelial dysfunctionand diabetic nephropathy.METHODS Primary glomerular endothelial cells were subjected to assess permeability under injury of advanced glycation end-products(AGEs).AGEs-induced changes of Rho A/ROCK pathway and cytoskeleton rearrangement were assessed bywestern blotandimmunofluorescence.The beneficial effects of SalA on diabetic nephropathy were investigated in a rat model induced by high-fat and high-glucose diet combined with low dose of streptozocin(35 mg·kg^(-1),ip).Renal function and architectonic changes were evaluated by biochemical assay and PAS staining.RESULTS SalA 3μMameliorated AGEs-induced glomerular endothelial permeability(P<0.05)and suppressed rearrangement of cytoskeleton through inhibiting AGE-RAGE-Rho A/ROCK pathway.SalA1 mg·kg^(-1)markedly reduced endothelium loss(P<0.01)and glomerular hyperfiltration(P<0.05)in diabetic kidney.Subsequently,SalA 1 mg·kg^(-1) suppressed glomerular hypertrophy and mesangial matrix expansion,eventually reduced 24 h-urinary albumin and ameliorated renal function by decreasing blood urine nitrogen(BUN),serum creatinine(Scr)and serum n-acetyl-β-d-glucosaminidase(NAG).AGEs-RAGE-Nox4-induced oxidative stress was suppressed by the treatment of SalA 1 mg·kg^(-1).CONCLUSION SalA ameliorated AGEs-induced glomerular endothelial hyperpermeability,and effectively protected against early-stage diabetic nephropathy by reducing hyperfiltration and alleviating renal structural deterioration through inhibiting AGEs and its downstream pathway.Thus,SalA might be a promising therapeutic agent for the treatment of diabetic nephropathy.展开更多
OBJECTIVE To investigate possible protective effect of berberine,an isoquinoline alkaloid,is the major active constituent of Rhizoma coptidis and Cortex phellodendri,on high-fat diet/streptozotocin-induced early diabe...OBJECTIVE To investigate possible protective effect of berberine,an isoquinoline alkaloid,is the major active constituent of Rhizoma coptidis and Cortex phellodendri,on high-fat diet/streptozotocin-induced early diabetic nephropathy in rats and various mechanisms underlie this effect.METHODS The diabetic rat model was generated by a single intraperitoneal injection of streptozotocin(STZ,50 mg·kg-1).Diabetic rats were randomlyassigned into the following five groups:control,DN,losartan(30 mg·kg-1·d-1),berberine(100,200 mg·kg-1·d-1).Berberine and losartan were given intragastricly for nine weeks.At the end of the experiment,urine of each group was collected in a 24 h period.Rats were weighed and then sacrificed.Plasma and kidneys were collected.The levels of blood glucose,creatinine(Cr),triglycerides(TG),total cholesterol(TCH)and malondialdehyde(MDA)in serum were determined using commercial kits according to the manufacturer′s instructions.Transforming growth factor(TGF)-β1and intracellular adhesion molecule-1(IAM-1)mR NA levels were evaluated by RT-PCR.The renal histopathology was observed by light microscopy.Further biochemical analysis of IKKβ1 and p65(nucleus/cytoplasm)was provided using Western blotting techniques.RESULTS Our study has demonstrated that berberine has various pharmacological activities.The DN rats had significantly higher kidney/body weight ratio(17.4±1.4)mg·g-1,and berberine treatment could reduce this ratio change 13.6±0.6 and(11.6±0.8)mg·g-1,respectively;glucose control still remains the only disease-modifying therapy for diabetic complications,FBG was also recorded in the experiment.The findings reveal that the DN group showed a significantly higher glucose level(28.67±2.78)mmol·L-1.Treatment with8 weeks of berberine improved these parameters except blood glucose〔(18.67±2.59)mmol·L-1and 16.45±1.80 vs(28.67±2.78)mmol·L-1:plasma levels of urea nitrogen(15.67±2.48)mmol·L-1and 14.45±2.40 vs(12.26±2.40)mmol·L-1〕;plasma levels of 24 h urine albuminuria〔30.48±1.56 and 25.72±2.24 vs(15.26±0.12)μg·d-1〕;based on these results,berberine supposed to improve renal functions in diabetic rats.Berberine also ameliorated the inflammatory changes of DN in diabetic animals;levels of TG,TCH and MDA in berberine-treatment rats were significantly lower compared with those in the DN group:TG〔2.78±0.24 and 2.45±0.36 vs(5.20±0.60)mmol·L-1〕;TCH〔4.26±0.46 and 3.74±0.68 vs(6.26±0.50)mmol·L-1〕;MDA〔4.94±1.19 and 4.28±0.64 vs(4.28±0.64)nu·mL-1〕.Chronic inflammation,as is observed in diabetes,is associated with increased production of TGF-β1and IAM-1.Compared with the renal tissues of DN group,TGF-β1and IAM-1 gene expressions in berberine treated groups were reduced at the dose levels(100 and 200 mg·kg-1).And TGF-β1and IAM-1levels in berberine treated groups were reduced in a dosedependent manner:Relative expression of TGF-β1mR NA level(3.56±0.28 and 3.12±0.14 vs 5.12±0.44);Relative expression of IAM-1 mR NA leve(l1.78±0.56 and 1.42±0.24vs 4.36±0.35).Research finds that the NF-κB signaling pathway is activated in the renal tissue of diabetic mice and berberine inhibits activation of the NF-κB pathway:staining score for IKKβ1(4.34±0.26 and 3.82±0.24 vs 6.23±0.76),staining score for p65(2.34±0.26 and 1.74±0.78 vs 6.23±0.24)in nucleus and staining score for p65(7.21±0.13 and8.15±0.45 vs 4.23±0.54)in cytoplasm.CONCLUSION In this field,berberine suppresses the increased expression of p65 in the nucleus and decreases it in cytoplasm,which leads to the inhibition of the NF-κB pathway.These changes will result in decreasing the transcription and translation of many inflammatory mediators,such as TGF-β1and IAM-1.Additionally,these changes decrease the number of inflammatory cells and mononuclear macrophage infiltration into glomeruli and renal interstitium.These results indicated that berberine can protect the kidney of STZ-diabetic rats by reducing the expression of TGF-β1and IAM-1 in the renal tubulointerstitium.And we propose that berberinemayfunction as an effective therapeutic agent for diabetic nephropathy and attenuate the progression of renal injury.展开更多
Diabetic nephropathy(DN)is one of the most common complications of diabetes.It is an important cause of diabetes disability and death.DN is a systemic metabolic syndrome.In its pathogenesis,the interaction of various ...Diabetic nephropathy(DN)is one of the most common complications of diabetes.It is an important cause of diabetes disability and death.DN is a systemic metabolic syndrome.In its pathogenesis,the interaction of various cell activities and a large number of cytokine biological activities,the activation of signal pathways and so on are involved in the development of DN.At present,the clinical treatment of DN is mainly Western medicine,but it has limitations such as strong toxicity,high side effects and poor compliance.Therefore,the discovery of natural anti-DN substances has also become an important means to treat DN.Mulberry leaves are the dry leaves of Morus alba L.It is not only a traditional Chinese medicine,but also a dual-purpose medicinal material for medicine and food.It has the effects of dispelling wind and clearing heat,cooling blood and brightening eyes,tonifying and so on.Mulberry leaf polysaccharide(MLP)is a kind of high molecular compound in mulberry leaves.It has many pharmacological effects,such as hypoglycemic,antioxidant,anti-stress,anti-virus and so on.Therefore,the pharmacological effects of mulberry leaf polysaccharides on diabetic nephropathy are reviewed in this paper,so as to provide references for further research and application.The pathogenesis of DN is complex,and the mechanism of renal injury has not been completely clarified.The current studies believe that DN is closely related to heredity,abnormal glucose metabolism,abnormal lipid metabolism,microcirculation disorder,cytokine action,oxidative stress and so on.Relevant studies show that the pharmacological effects of mulberry leaf polysaccharide in the prevention and treatment of DN mainly include:①Effect on transforming factor-β1(TGF-β1):TGF-β1 has become an important cytokine involved in the formation of renal fibrosis by regulating cell proliferation and differentiation and the production of extracellular matrix(ECM).MLP can significantly inhibit TGF-β1 protein,and then inhibit the synthesis of extracellular matrix by renal interstitial fibroblasts and inhibit the realization of fibrosis.②Effect on insulin receptor substrate(IRS-1):IRS-1 is an important signal molecule at the beginning of IR signal transduction.The decrease of IRS-1 gene expression or the decrease of expression can affect the effective transmission of IR signal and lead to the development and deterioration of diabetes. MPL can significantly increase the expression of IRS-1 mRNA in liver tissue of DN rats, so as to prevent and treat DN. ③ Effect on the expression of resistin protein in adipose tis sue. Resistin is a secretory polypeptide derived from adipose tissue and is specifically expressed in white adipose tissue and is closely related to type 2 diabetes mellitus (T2DM). Experimental studies show that MLP can effectively reduce the expression of resistin protein in white adipose tissue of T2DM rats, indicating that MLP may reduce the level of IR by inhibiting the expression of resistin in adipose tissue, thereby reducing the insulin resistance state of T2DM rats, so as to achieve the goal of treating diabetes. ④ Effect on adiponectin receptor 1 (AdipoR1): adiponectin can improve insulin resistance, reduce blood glucose and lipid. AdipoR1 is mainly expressed in skeletal muscle and kidney. Studies have shown that AdipoR1 is closely related to the occurrence and development of DN. The results showed that MLP could reduce the blood glucose and blood lipid level and up regulate the expression of AdipoR1 mRNA in DN rats, suggesting that MLP may delay the occurrence and development of DN. This article reviewed the pharmacological effects of mulberry leaf polysaccharides on diabetic nephropathy, and provided a useful basis for further development and utilization of mul berry leaf polysaccharides in the treatment of DN.展开更多
OBJECTIVE To observe the effect of Qi Kwai Granule particles on the expression of in.terleukin 6(IL-6),monocyte chemotactic protein 1(MCP-1) and transforming growth factor-β1(TGF-β1)in diabetic nephropathy(DN) rats ...OBJECTIVE To observe the effect of Qi Kwai Granule particles on the expression of in.terleukin 6(IL-6),monocyte chemotactic protein 1(MCP-1) and transforming growth factor-β1(TGF-β1)in diabetic nephropathy(DN) rats and evaluate the protective effect of Qi Kwai Granule particles against renal injury of diabetic nephropathy.METHODS This experiment adopts adopted the high-sugar-highfat diet and intraperitoneal injection of 2% STZ+ unilateral renal ligation to establish rat model of diabet.ic nephropathy.50 model rats were then randomly divided into model group,Irbesartan group,Qi Kwai Granule particles of high,medium,low dose group,10 rats in each group.10 normal rats were set as the sham operation group.Intragastric administration for 8 weeks were measured in rats.Measure the value of rat blood glucose by blood glucose meter,the determination of serum interleukin 6(IL-6) con.tent by ELISA,the expression of MCP-1 and TGF-β1 by immunohistochemistry method.The value of rat blood glucose were measured by blood glucose meter.Serum interleukin 6(IL-6) were determinat.ed by ELISA.Expression of MCP-1 and TGF-β1 were evaluated by immunohistochemistry method.RE.SULTS The blood glucose of Qi Kwai Granule particles of high,medium groups were decreased com.pared with those of the model group(P<0.05).The content of IL-6 of Qi Kwai Granule particles of high,medium groups were reduced(P<0.01).The content of MCP-1,TGF-β1 in kidney of Qi Kwai Granule particles of high,medium,low dose groups were decreased(P<0.01).CONCLUSION Qi Kwai parti.cles have protective effect on renal tissue of diabetic nephropathy rats.Its mechanism might be related to the decrease of blood glucose value and IL-6,the inhibition of the expression of MCP-1 and TGF-β1.展开更多
Aim To investigate the effects of Ginkgo biloba extract （GbE） on renal fibrosis in STZ induced diabetic rats and high glucose （HG） cultured proximal tubular epithelial cells （NRK-52E）. Methods In vivo, rats were...Aim To investigate the effects of Ginkgo biloba extract （GbE） on renal fibrosis in STZ induced diabetic rats and high glucose （HG） cultured proximal tubular epithelial cells （NRK-52E）. Methods In vivo, rats were randomized into six groups termed normal control, diabetes mellitus , low dose of GbE （50 mg · kg^-1 · d^-1） , in- termediate dose of GbE （100 mg · kg^-1·d^-1）, high dose of GbE （200 mg · kg^-1 · d^-1） and rapamycin （1 mg·kg^-1·d^-l）. After 12 weeks, the rats were sacrificed and then fasting blood glucose, creatinine （Cr） , blood u- rea nitrogen （BUN） , urine protein, kidney index, glycogen and collagen accumulation, and collagen IV and lami- NRK-52E cells were divided into six groups： normal nin expression were measured by different methods. In vitro, glucose （5.56 mmol · L^-1）, high glucose （60 mmol · L^-1）, low dose of GbE （10 mg · L^-1）, intermediate dose of GbE （20 mg· L^-1）, high dose of GbE （40 mg· L^-1） and rapamycin （20 nmol · L^-l）. The morphological changes of cells were observed by microscopy after culturing for 48 h. Akt, roTOR and p70S6K, were examined by western blotting both in the renal cortex of rats and NRK-52E cells. Results Compared with diabetic rats, the lev- els of Cr, BUN, urine protein, kidney index, accumulation of glycogen and collagen, and expression of collagen IV and laminin in the renal cortex were all decreased in GbE treated rats. Furthermore, GbE ameliorated the morpho- logical changes of NRK-52E cells caused by HG. In addition, GbE reduced the expression of E-cadherin, oL-SMA, snail and phosphorylation of Akt, roTOR and p70S6K in diabetic renal cortexes and NRK-52E cells exposed to HG. Conclusion GbE was a satisfactory agent to prevent renal fibrosis in diabetic nephropathy, and this effect might be associated with the inhibition of the Akt/mTOR signaling pathway.展开更多
Aim The study was aimed to investigate the effect of Huangqi decoction （HD） on diabetic nephropa- thy. Methods Male diabetic db/db mice which develop diabetic nephropathy spontanously and no-diabetic db/m control mi...Aim The study was aimed to investigate the effect of Huangqi decoction （HD） on diabetic nephropa- thy. Methods Male diabetic db/db mice which develop diabetic nephropathy spontanously and no-diabetic db/m control mice were used in the current study, and they received the treatment of HD for 14 consecutive weeks. Re- sults HD treatment dose-dependently decreased the body weight, urine volume, water intake, food intake in the db/db mice, improved glucose tolerance and insulin resistance, lowered blood glucose, systolic blood pressure, se- rum glyeosylated hemoglobin, insulin and insulin resistance index. The db/db mice also showed low levels of serum and urine creatinine, blood urea nitrogen and urine protein, and improved renal functions such as glomerular filtra- tion rate and ATP production after HD treatment. Histological examination by PAS staining showed that HD treat- ment prevented the deterioration of basement membrane of glomerular capillary, mesangial matrix and renal tubular lumen in the db/db mice. Examining the cell signaling pathways which might be involved the pathology of diabe- tes, we found that HD up-regulated the expressions of phospho-IR, phospho-IRS1307 phospho-PI3K and GLUT4 and down-regulated the expression of phospho-IRS1636, phospho-AKT308, phospho-AKT473 and GLUT1 in a dose-de- pendent manner. Conclusion Our study suggests that HD regulates the IRS1-PI3 K-GLUT signaling pathway and significantly improves diabetic nephropathy.展开更多
Objective Apoptosis is recognized as an important mechanism in contrast-induced nephropathy(CIN).Acupuncture and moxibustion,the auxiliary treatment in China,are effective interventions for cell apoptosis in many isch...Objective Apoptosis is recognized as an important mechanism in contrast-induced nephropathy(CIN).Acupuncture and moxibustion,the auxiliary treatment in China,are effective interventions for cell apoptosis in many ischemic diseases.In our previous study,we found acupuncture and moxibustion could prevent CIN.The objective of this research is to study the mechanism of acupuncture and moxibustion on tubular epithelial cell apoptosis in diabetic CIN rats.展开更多
目的:探究尿液细胞外囊泡中关键蛋白细胞骨架相关蛋白4(cytoskeleton⁃associated protein 4,CKAP4)作为生物标志物预测糖尿病肾病(diabetic kidney disease,DKD)进展的能力。方法:纳入南京医科大学第一附属医院143例肾活检证实DKD的2型...目的:探究尿液细胞外囊泡中关键蛋白细胞骨架相关蛋白4(cytoskeleton⁃associated protein 4,CKAP4)作为生物标志物预测糖尿病肾病(diabetic kidney disease,DKD)进展的能力。方法:纳入南京医科大学第一附属医院143例肾活检证实DKD的2型糖尿病患者以及肾脏恶性肿瘤患者10例。免疫组化法检测肾组织中CKAP4的表达水平。采用Spearman相关分析CKAP4表达水平与临床指标的相关性。计算受试者工作特征(receiver operating characteristic,ROC)曲线下面积(area under the curve,AUC),以评估CKAP4表达是否能有效区分肾脏预后不良的患者。采用ROC曲线分析确定预测肾脏事件的CKAP4评分的最佳临界值(最高约登指数)。此外,进行时间依赖性AUC分析,以表征肾活检后6个月以上CKAP4的预测准确性。通过Cox比例风险模型随访CKAP4和DKD进展之间的风险比。将单变量分析中有统计学意义(P<0.05)的临床病理因素作为协变量纳入多变量Cox比例风险模型分析。采用Kaplan⁃Meier分析评价CKAP4高表达组和CKAP4低表达组肾活检后6个月以上生存的差异。结果:与肿瘤患者癌旁正常肾脏组织相比,CKAP4在DKD患者肾脏组织中表达增高,差异有统计学意义(P<0.001)。CKAP4在不同分期的DKD患者肾组织中表达有差异,其中Ⅱ期与Ⅲ期、Ⅱ期与Ⅳ期、Ⅲ期与Ⅳ期之间的差异均有统计学意义(P均<0.05)。DKD患者CKAP4表达与血清肌酐、尿素氮、24 h尿蛋白呈正相关,与肾小球滤过率估计值、血红蛋白呈负相关。DKD患者中位随访期为2.22年,63例(44.06%)患者出现DKD进展。Pearson相关分析显示,CKAP4表达水平随DKD病理分级的升高而升高(r=0.808,P<0.001)。多因素Cox回归分析显示,CKAP4高表达与DKD进展的风险增加相关(HR=4.120,95%CI:1.730~9.811,P=0.001)。此外,在Kaplan⁃Meier生存分析中,CKAP4高表达组患者的肾脏终点事件的发生率显著高于CKAP4低表达组(P<0.001)。绘制了包括CKAP4分类在内的列线图来预测DKD进展风险(C⁃index:0.689)。结论:来源于尿液细胞外囊泡的CKAP4是DKD患者肾活检后6个月以上疾病进展的独立危险因素。展开更多
文摘Aim To investigate the nephroprotective activity of berberine in diabetic nephropathy （DN） mice. Methods Diabetic nephropathy was induced by intraperitoneal injection with 55 mg · kg^-1 streptozotocin（STZ） , Berberine was administered at daily doses of 50, 100 and 200 mg· kg^-1 by gavage for 8 weeks. To detect serum creatinine and blood urea nitrogen （BUN） levels, blood were collected after the last dose of berberine, renal cortex was separated on ice after heart peffusion by precooled normal saline. The specimen was stored in -80℃ for the next experiments, and some of the kidney tissue were immobilized by 4% paraformaldehyde solution and 3% glut- araldehyde solution for the preparation of paraffin tissue slides and electron microscope biopsy respectively. After that, PAS staining and electron microscope were used to observe the glomerular morphology changes; ELISA was used to measure proinflammatory chemokines and cytokines levels in renal cortex. Real time RT-PCR was taken to detect the level of nucleotide binding oligomerzation domain 2 （NOD2） mRNA, Western blot was used to test the ex- pression of NOD2 and autophagy marker light chain 3 （LC3） in renal cortex. Results Histopathological changes and the increase in serum creatinine and BUN in DN mice were significantly ameliorated by berberine in a dose-de- pendent manner. Additionally, The expression of tumor necrosis factor-or （TNF-α）, interleukin-6 （IL-6） and in- tercellular adhesion molecule-1 （ICAM-1） was markedly suppressed by berberine, indicating the inhibition of in- flammatory response. Treatment of DN mice with berberine also significantly reduced the expression of NOD2 and LC3 in the kidneys. Conclusion The current study showed the nephroprotective activity of berberine in DN mice could be attributed to the inhibition of inflammation and
基金The project supported by Ministry Education of Indonesia
文摘OBJECTIVE To investigate the effect of Urena lobataleaves extract on the inhibition of nephropathy diabetic complication.METHODS This study uses control group post test only with male Sprague dawley rats.Diabetic rats was induced by high fructose diet(HFD)and single dose streptozotocin 25mg·kg-1 bw intra peritoneal.The rat was administrated orally with water extract of U.lobataleaves in concentrations of 250,500 and 1000mg·kg-1 bw for 4 weeks.After scarifying,kidney organ were collected and then superoxyde dismutase(SOD)kidney level,malondialdehyda(MDA)and tumor necrosis factor-alpha(TNF-α)were examined.The data was analyzed using ANOVA test continued with LSD test(P<0.05).RESULTS The oral administration of U.lobataleaves extract 250,500 and 1000mg·kg-1 bw were able to increase SOD kidney level about 30%,60% and 90% respectively compared to diabetic group(P<0.05),while the MDA kidney level was decreased by 30%,60% and 70%(P<0.05)respectively.The supplementation of water extract from U.lobatain dose of 250,500 and 1000mg·kg-1 bw were also decrease TNF-αkidney level approximately 30%,40% and 60% compared to control group(P<0.05).In diabetic groups,SOD kidney level was decreased compared to normal group(P<0.05)while the MDA and TNF-αwere increased(P<0.05).CONCLUSION U.lobata leaves extract could inhibit nephropathy diabetic complication by increasing of SOD kidney level,decreasing of MDA kidney level,and TNF-α.This effect may be related to active compounds that act as an antioxidant and anti-inflammatory in U.lobata extract.
基金supported by National Nature Science Foundation of China(81770847)CAMS Innovation Fund for Medical Sciences(CIFMS)(2016-I2M-3-007,2016-I2M-1-010)National Key Research and Development Plan(2016YFC1000905)
文摘OBJECTIVE Diabetic nephropathy(DN)has been one of the most common complications of diabetes and the leading cause of end-stage renal disease.Glomerular hyperfiltrationis central in earlystage of DN and leads to the progression of renal architectonic and functional abnormalities.Salvianolic acid A(SalA)has been proved to protect diabetic complications such as hepatic fibrosis and neuropathy.The present study was designed to investigate the effects of SalA on glomerular endothelial dysfunctionand diabetic nephropathy.METHODS Primary glomerular endothelial cells were subjected to assess permeability under injury of advanced glycation end-products(AGEs).AGEs-induced changes of Rho A/ROCK pathway and cytoskeleton rearrangement were assessed bywestern blotandimmunofluorescence.The beneficial effects of SalA on diabetic nephropathy were investigated in a rat model induced by high-fat and high-glucose diet combined with low dose of streptozocin(35 mg·kg^(-1),ip).Renal function and architectonic changes were evaluated by biochemical assay and PAS staining.RESULTS SalA 3μMameliorated AGEs-induced glomerular endothelial permeability(P<0.05)and suppressed rearrangement of cytoskeleton through inhibiting AGE-RAGE-Rho A/ROCK pathway.SalA1 mg·kg^(-1)markedly reduced endothelium loss(P<0.01)and glomerular hyperfiltration(P<0.05)in diabetic kidney.Subsequently,SalA 1 mg·kg^(-1) suppressed glomerular hypertrophy and mesangial matrix expansion,eventually reduced 24 h-urinary albumin and ameliorated renal function by decreasing blood urine nitrogen(BUN),serum creatinine(Scr)and serum n-acetyl-β-d-glucosaminidase(NAG).AGEs-RAGE-Nox4-induced oxidative stress was suppressed by the treatment of SalA 1 mg·kg^(-1).CONCLUSION SalA ameliorated AGEs-induced glomerular endothelial hyperpermeability,and effectively protected against early-stage diabetic nephropathy by reducing hyperfiltration and alleviating renal structural deterioration through inhibiting AGEs and its downstream pathway.Thus,SalA might be a promising therapeutic agent for the treatment of diabetic nephropathy.
基金The project supported by Fundamental Research Funds for the Central Universities(21615463)
文摘OBJECTIVE To investigate possible protective effect of berberine,an isoquinoline alkaloid,is the major active constituent of Rhizoma coptidis and Cortex phellodendri,on high-fat diet/streptozotocin-induced early diabetic nephropathy in rats and various mechanisms underlie this effect.METHODS The diabetic rat model was generated by a single intraperitoneal injection of streptozotocin(STZ,50 mg·kg-1).Diabetic rats were randomlyassigned into the following five groups:control,DN,losartan(30 mg·kg-1·d-1),berberine(100,200 mg·kg-1·d-1).Berberine and losartan were given intragastricly for nine weeks.At the end of the experiment,urine of each group was collected in a 24 h period.Rats were weighed and then sacrificed.Plasma and kidneys were collected.The levels of blood glucose,creatinine(Cr),triglycerides(TG),total cholesterol(TCH)and malondialdehyde(MDA)in serum were determined using commercial kits according to the manufacturer′s instructions.Transforming growth factor(TGF)-β1and intracellular adhesion molecule-1(IAM-1)mR NA levels were evaluated by RT-PCR.The renal histopathology was observed by light microscopy.Further biochemical analysis of IKKβ1 and p65(nucleus/cytoplasm)was provided using Western blotting techniques.RESULTS Our study has demonstrated that berberine has various pharmacological activities.The DN rats had significantly higher kidney/body weight ratio(17.4±1.4)mg·g-1,and berberine treatment could reduce this ratio change 13.6±0.6 and(11.6±0.8)mg·g-1,respectively;glucose control still remains the only disease-modifying therapy for diabetic complications,FBG was also recorded in the experiment.The findings reveal that the DN group showed a significantly higher glucose level(28.67±2.78)mmol·L-1.Treatment with8 weeks of berberine improved these parameters except blood glucose〔(18.67±2.59)mmol·L-1and 16.45±1.80 vs(28.67±2.78)mmol·L-1:plasma levels of urea nitrogen(15.67±2.48)mmol·L-1and 14.45±2.40 vs(12.26±2.40)mmol·L-1〕;plasma levels of 24 h urine albuminuria〔30.48±1.56 and 25.72±2.24 vs(15.26±0.12)μg·d-1〕;based on these results,berberine supposed to improve renal functions in diabetic rats.Berberine also ameliorated the inflammatory changes of DN in diabetic animals;levels of TG,TCH and MDA in berberine-treatment rats were significantly lower compared with those in the DN group:TG〔2.78±0.24 and 2.45±0.36 vs(5.20±0.60)mmol·L-1〕;TCH〔4.26±0.46 and 3.74±0.68 vs(6.26±0.50)mmol·L-1〕;MDA〔4.94±1.19 and 4.28±0.64 vs(4.28±0.64)nu·mL-1〕.Chronic inflammation,as is observed in diabetes,is associated with increased production of TGF-β1and IAM-1.Compared with the renal tissues of DN group,TGF-β1and IAM-1 gene expressions in berberine treated groups were reduced at the dose levels(100 and 200 mg·kg-1).And TGF-β1and IAM-1levels in berberine treated groups were reduced in a dosedependent manner:Relative expression of TGF-β1mR NA level(3.56±0.28 and 3.12±0.14 vs 5.12±0.44);Relative expression of IAM-1 mR NA leve(l1.78±0.56 and 1.42±0.24vs 4.36±0.35).Research finds that the NF-κB signaling pathway is activated in the renal tissue of diabetic mice and berberine inhibits activation of the NF-κB pathway:staining score for IKKβ1(4.34±0.26 and 3.82±0.24 vs 6.23±0.76),staining score for p65(2.34±0.26 and 1.74±0.78 vs 6.23±0.24)in nucleus and staining score for p65(7.21±0.13 and8.15±0.45 vs 4.23±0.54)in cytoplasm.CONCLUSION In this field,berberine suppresses the increased expression of p65 in the nucleus and decreases it in cytoplasm,which leads to the inhibition of the NF-κB pathway.These changes will result in decreasing the transcription and translation of many inflammatory mediators,such as TGF-β1and IAM-1.Additionally,these changes decrease the number of inflammatory cells and mononuclear macrophage infiltration into glomeruli and renal interstitium.These results indicated that berberine can protect the kidney of STZ-diabetic rats by reducing the expression of TGF-β1and IAM-1 in the renal tubulointerstitium.And we propose that berberinemayfunction as an effective therapeutic agent for diabetic nephropathy and attenuate the progression of renal injury.
文摘Diabetic nephropathy(DN)is one of the most common complications of diabetes.It is an important cause of diabetes disability and death.DN is a systemic metabolic syndrome.In its pathogenesis,the interaction of various cell activities and a large number of cytokine biological activities,the activation of signal pathways and so on are involved in the development of DN.At present,the clinical treatment of DN is mainly Western medicine,but it has limitations such as strong toxicity,high side effects and poor compliance.Therefore,the discovery of natural anti-DN substances has also become an important means to treat DN.Mulberry leaves are the dry leaves of Morus alba L.It is not only a traditional Chinese medicine,but also a dual-purpose medicinal material for medicine and food.It has the effects of dispelling wind and clearing heat,cooling blood and brightening eyes,tonifying and so on.Mulberry leaf polysaccharide(MLP)is a kind of high molecular compound in mulberry leaves.It has many pharmacological effects,such as hypoglycemic,antioxidant,anti-stress,anti-virus and so on.Therefore,the pharmacological effects of mulberry leaf polysaccharides on diabetic nephropathy are reviewed in this paper,so as to provide references for further research and application.The pathogenesis of DN is complex,and the mechanism of renal injury has not been completely clarified.The current studies believe that DN is closely related to heredity,abnormal glucose metabolism,abnormal lipid metabolism,microcirculation disorder,cytokine action,oxidative stress and so on.Relevant studies show that the pharmacological effects of mulberry leaf polysaccharide in the prevention and treatment of DN mainly include:①Effect on transforming factor-β1(TGF-β1):TGF-β1 has become an important cytokine involved in the formation of renal fibrosis by regulating cell proliferation and differentiation and the production of extracellular matrix(ECM).MLP can significantly inhibit TGF-β1 protein,and then inhibit the synthesis of extracellular matrix by renal interstitial fibroblasts and inhibit the realization of fibrosis.②Effect on insulin receptor substrate(IRS-1):IRS-1 is an important signal molecule at the beginning of IR signal transduction.The decrease of IRS-1 gene expression or the decrease of expression can affect the effective transmission of IR signal and lead to the development and deterioration of diabetes. MPL can significantly increase the expression of IRS-1 mRNA in liver tissue of DN rats, so as to prevent and treat DN. ③ Effect on the expression of resistin protein in adipose tis sue. Resistin is a secretory polypeptide derived from adipose tissue and is specifically expressed in white adipose tissue and is closely related to type 2 diabetes mellitus (T2DM). Experimental studies show that MLP can effectively reduce the expression of resistin protein in white adipose tissue of T2DM rats, indicating that MLP may reduce the level of IR by inhibiting the expression of resistin in adipose tissue, thereby reducing the insulin resistance state of T2DM rats, so as to achieve the goal of treating diabetes. ④ Effect on adiponectin receptor 1 (AdipoR1): adiponectin can improve insulin resistance, reduce blood glucose and lipid. AdipoR1 is mainly expressed in skeletal muscle and kidney. Studies have shown that AdipoR1 is closely related to the occurrence and development of DN. The results showed that MLP could reduce the blood glucose and blood lipid level and up regulate the expression of AdipoR1 mRNA in DN rats, suggesting that MLP may delay the occurrence and development of DN. This article reviewed the pharmacological effects of mulberry leaf polysaccharides on diabetic nephropathy, and provided a useful basis for further development and utilization of mul berry leaf polysaccharides in the treatment of DN.
文摘OBJECTIVE To observe the effect of Qi Kwai Granule particles on the expression of in.terleukin 6(IL-6),monocyte chemotactic protein 1(MCP-1) and transforming growth factor-β1(TGF-β1)in diabetic nephropathy(DN) rats and evaluate the protective effect of Qi Kwai Granule particles against renal injury of diabetic nephropathy.METHODS This experiment adopts adopted the high-sugar-highfat diet and intraperitoneal injection of 2% STZ+ unilateral renal ligation to establish rat model of diabet.ic nephropathy.50 model rats were then randomly divided into model group,Irbesartan group,Qi Kwai Granule particles of high,medium,low dose group,10 rats in each group.10 normal rats were set as the sham operation group.Intragastric administration for 8 weeks were measured in rats.Measure the value of rat blood glucose by blood glucose meter,the determination of serum interleukin 6(IL-6) con.tent by ELISA,the expression of MCP-1 and TGF-β1 by immunohistochemistry method.The value of rat blood glucose were measured by blood glucose meter.Serum interleukin 6(IL-6) were determinat.ed by ELISA.Expression of MCP-1 and TGF-β1 were evaluated by immunohistochemistry method.RE.SULTS The blood glucose of Qi Kwai Granule particles of high,medium groups were decreased com.pared with those of the model group(P<0.05).The content of IL-6 of Qi Kwai Granule particles of high,medium groups were reduced(P<0.01).The content of MCP-1,TGF-β1 in kidney of Qi Kwai Granule particles of high,medium,low dose groups were decreased(P<0.01).CONCLUSION Qi Kwai parti.cles have protective effect on renal tissue of diabetic nephropathy rats.Its mechanism might be related to the decrease of blood glucose value and IL-6,the inhibition of the expression of MCP-1 and TGF-β1.
文摘Aim To investigate the effects of Ginkgo biloba extract （GbE） on renal fibrosis in STZ induced diabetic rats and high glucose （HG） cultured proximal tubular epithelial cells （NRK-52E）. Methods In vivo, rats were randomized into six groups termed normal control, diabetes mellitus , low dose of GbE （50 mg · kg^-1 · d^-1） , in- termediate dose of GbE （100 mg · kg^-1·d^-1）, high dose of GbE （200 mg · kg^-1 · d^-1） and rapamycin （1 mg·kg^-1·d^-l）. After 12 weeks, the rats were sacrificed and then fasting blood glucose, creatinine （Cr） , blood u- rea nitrogen （BUN） , urine protein, kidney index, glycogen and collagen accumulation, and collagen IV and lami- NRK-52E cells were divided into six groups： normal nin expression were measured by different methods. In vitro, glucose （5.56 mmol · L^-1）, high glucose （60 mmol · L^-1）, low dose of GbE （10 mg · L^-1）, intermediate dose of GbE （20 mg· L^-1）, high dose of GbE （40 mg· L^-1） and rapamycin （20 nmol · L^-l）. The morphological changes of cells were observed by microscopy after culturing for 48 h. Akt, roTOR and p70S6K, were examined by western blotting both in the renal cortex of rats and NRK-52E cells. Results Compared with diabetic rats, the lev- els of Cr, BUN, urine protein, kidney index, accumulation of glycogen and collagen, and expression of collagen IV and laminin in the renal cortex were all decreased in GbE treated rats. Furthermore, GbE ameliorated the morpho- logical changes of NRK-52E cells caused by HG. In addition, GbE reduced the expression of E-cadherin, oL-SMA, snail and phosphorylation of Akt, roTOR and p70S6K in diabetic renal cortexes and NRK-52E cells exposed to HG. Conclusion GbE was a satisfactory agent to prevent renal fibrosis in diabetic nephropathy, and this effect might be associated with the inhibition of the Akt/mTOR signaling pathway.
文摘Aim The study was aimed to investigate the effect of Huangqi decoction （HD） on diabetic nephropa- thy. Methods Male diabetic db/db mice which develop diabetic nephropathy spontanously and no-diabetic db/m control mice were used in the current study, and they received the treatment of HD for 14 consecutive weeks. Re- sults HD treatment dose-dependently decreased the body weight, urine volume, water intake, food intake in the db/db mice, improved glucose tolerance and insulin resistance, lowered blood glucose, systolic blood pressure, se- rum glyeosylated hemoglobin, insulin and insulin resistance index. The db/db mice also showed low levels of serum and urine creatinine, blood urea nitrogen and urine protein, and improved renal functions such as glomerular filtra- tion rate and ATP production after HD treatment. Histological examination by PAS staining showed that HD treat- ment prevented the deterioration of basement membrane of glomerular capillary, mesangial matrix and renal tubular lumen in the db/db mice. Examining the cell signaling pathways which might be involved the pathology of diabe- tes, we found that HD up-regulated the expressions of phospho-IR, phospho-IRS1307 phospho-PI3K and GLUT4 and down-regulated the expression of phospho-IRS1636, phospho-AKT308, phospho-AKT473 and GLUT1 in a dose-de- pendent manner. Conclusion Our study suggests that HD regulates the IRS1-PI3 K-GLUT signaling pathway and significantly improves diabetic nephropathy.
文摘Objective Apoptosis is recognized as an important mechanism in contrast-induced nephropathy(CIN).Acupuncture and moxibustion,the auxiliary treatment in China,are effective interventions for cell apoptosis in many ischemic diseases.In our previous study,we found acupuncture and moxibustion could prevent CIN.The objective of this research is to study the mechanism of acupuncture and moxibustion on tubular epithelial cell apoptosis in diabetic CIN rats.
文摘目的:探究尿液细胞外囊泡中关键蛋白细胞骨架相关蛋白4(cytoskeleton⁃associated protein 4,CKAP4)作为生物标志物预测糖尿病肾病(diabetic kidney disease,DKD)进展的能力。方法:纳入南京医科大学第一附属医院143例肾活检证实DKD的2型糖尿病患者以及肾脏恶性肿瘤患者10例。免疫组化法检测肾组织中CKAP4的表达水平。采用Spearman相关分析CKAP4表达水平与临床指标的相关性。计算受试者工作特征(receiver operating characteristic,ROC)曲线下面积(area under the curve,AUC),以评估CKAP4表达是否能有效区分肾脏预后不良的患者。采用ROC曲线分析确定预测肾脏事件的CKAP4评分的最佳临界值(最高约登指数)。此外,进行时间依赖性AUC分析,以表征肾活检后6个月以上CKAP4的预测准确性。通过Cox比例风险模型随访CKAP4和DKD进展之间的风险比。将单变量分析中有统计学意义(P<0.05)的临床病理因素作为协变量纳入多变量Cox比例风险模型分析。采用Kaplan⁃Meier分析评价CKAP4高表达组和CKAP4低表达组肾活检后6个月以上生存的差异。结果:与肿瘤患者癌旁正常肾脏组织相比,CKAP4在DKD患者肾脏组织中表达增高,差异有统计学意义(P<0.001)。CKAP4在不同分期的DKD患者肾组织中表达有差异,其中Ⅱ期与Ⅲ期、Ⅱ期与Ⅳ期、Ⅲ期与Ⅳ期之间的差异均有统计学意义(P均<0.05)。DKD患者CKAP4表达与血清肌酐、尿素氮、24 h尿蛋白呈正相关,与肾小球滤过率估计值、血红蛋白呈负相关。DKD患者中位随访期为2.22年,63例(44.06%)患者出现DKD进展。Pearson相关分析显示,CKAP4表达水平随DKD病理分级的升高而升高(r=0.808,P<0.001)。多因素Cox回归分析显示,CKAP4高表达与DKD进展的风险增加相关(HR=4.120,95%CI:1.730~9.811,P=0.001)。此外,在Kaplan⁃Meier生存分析中,CKAP4高表达组患者的肾脏终点事件的发生率显著高于CKAP4低表达组(P<0.001)。绘制了包括CKAP4分类在内的列线图来预测DKD进展风险(C⁃index:0.689)。结论:来源于尿液细胞外囊泡的CKAP4是DKD患者肾活检后6个月以上疾病进展的独立危险因素。
