摘要
目的:研究白虎汤对2型糖尿病大鼠的血糖、血脂代谢,血管重构的影响及其对胰岛素受体底物-1(IRS-1)/胞内磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B(Akt)信号通路的调节作用。方法:将90只大鼠随机分为正常组、模型组、白虎汤低、中、高剂量组及二甲双胍组,15只/组,除正常组大鼠外,其余大鼠采用腹腔注射链脲佐菌素法建立2型糖尿病模型。白虎汤低、中、高剂量组大鼠根据体质量分别灌胃给予大鼠白虎汤溶液,剂量分别为5,10,20 g·kg^(-1),二甲双胍组灌胃给予大鼠二甲双胍(100 mg·kg^(-1)),正常组及模型组给予等体积生理盐水,所有大鼠每日给药1次,连续给药12周。给药结束后测定各组大鼠空腹血糖,糖化血红蛋白,血清肿瘤坏死因子-α(TNF-α),白细胞介素-6(IL-6),白细胞介素-1β(IL-1β),总胆固醇(TC),甘油三酯(TG)及低密度脂蛋白胆固醇(LDL-C)水平,实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠肝脏固醇调节元件结合蛋白1C(SREBP1C),乙酰CoA羧化酶(ACC),脂肪酸合成酶基因(FASN)等脂质合成基因及肉毒碱棕榈酰基转移酶1A(CPT1A),酰基辅酶A氧化酶1(ACOX1),重组人中链酰基辅酶A脱氢酶(ACADM)等脂肪酸氧化基因的mRNA水平,蛋白免疫印迹法(Western blot)检测大鼠肝脏IRS-1,PI3K,Akt蛋白水平,苏木素-伊红(HE)染色观察大鼠胸主动脉血管组织病理学变化,划痕实验检测大鼠胸主动脉血管平滑肌细胞迁移能力。结果:与正常组比较,模型组大鼠空腹血糖、糖化血红蛋白、血清TNF-α,IL-6,IL-1β,TC,TG及LDL-C水平,肝脏脂质合成基因mRNA水平,大鼠胸主动脉血管平滑肌细胞迁移能力均明显升高(P<0.05),肝脏脂肪酸氧化基因mRNA水平及IRS-1,PI3K,Akt蛋白表达明显降低(P<0.05),大鼠胸主动脉血管壁厚度明显升高,差异具有统计学意义(P<0.05);与模型组比较,白虎汤各剂量组大鼠空腹血糖、糖化血红蛋白、血清TNF-α,IL-6,IL-1β,TC,TG及LDL-C水平,肝脏脂质合成基因mRNA水平,大鼠胸主动脉血管平滑肌细胞迁移能力均明显降低(P<0.05),肝脏脂肪酸氧化基因mRNA水平及IRS-1,PI3K,Akt蛋白水平均明显升高(P<0.05),大鼠胸主动脉组织病理学明显改善,且血管壁厚度明显降低,差异具有统计学意义(P<0.05)。结论:白虎汤能够降低2型糖尿病大鼠血糖、血脂及血清炎症因子水平,调节肝脏脂质代谢相关基因的表达,改善胸主动脉血管组织病理学及血管重构,其机制可能与调节IRS-1/PI3K/Akt信号通路有关。
Objective:To study the effect of Baihutang on blood glucose,blood lipid metabolism and vascular remodeling in type 2 diabetic rats and its regulation on insulin receptor substrate-1(IRS-1)/phosphatidylinositol-3 kinase(PI3 K)/protein kinase B(Akt)signal pathway. Method: The 90 rats were randomly divided into normal group,model group,Baihutang low,middle and high dose groups and metformin group,with 15 rats in each group. Except for normal group,the other rats were injected intraperitoneally with streptozotocin to establish the model of type 2 diabetes. The rats in the low,middle and high dose groups were given Baihutang formula granules of 5,10,20 g·kg^(-1) respectively according to their body weight. The positive control group was given metformin(100 mg·kg^(-1))by intragastric administration,while those in the control group and model group were given the same amount of normal saline once a day for 12 weeks. The levels of fasting blood glucose,glycosylated hemoglobin,serum tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1 β(IL-1β),total cholesterol(TC),triglyceride(TG)and low-density lipoprotein cholesterol(LDLC)were measured after administration. The levels of sterol regulatory element binding protein 1 C(SREBP1 C),acetyl CoA carboxylase(ACC),fatty acid synthase gene(FASN)and carnitine palmitoyl transferase 1 A(CPT1 A),acylcoa oxidase 1(ACOX1),recombinant human acylcoa dehydrogenase(ACADM)mRNA in liver of rats were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR),Western blot was used to detect the protein levels of IRS-1,PI3 K and Akt in liver of rats. Hematoxylin-eosin(HE)staining was used for histopathological examination of rat thoracic aortic vessels. The migration ability of vascular smooth muscle cells in rat thoracic aorta was detected by scratch test. Result: Compared with the normal group,the fasting blood glucose,glycosylated hemoglobin,serum TNF-α,IL-6,IL-1β,TC,TG and LDL-C levels,liver lipid synthesis gene mRNA level and vascular smooth muscle cell migration ability of thoracic aorta in model group were significantly higher than those in normal group(P<0.05),while fatty acid oxidation gene mRNA level and IRS-1,PI3 K,Akt protein level in liver were significantly decreased in model group(P<0.05). The vascular wall thickness of thoracic aorta increased significantly in rats(P<0.05).Compared with model group,the levels of fasting blood glucose,glycosylated hemoglobin,serum TNF-α,IL-6,IL-1β,TC,TG and LDL-C,the level of lipid synthesis gene mRNA in liver and the migration ability of vascular smooth muscle cells in thoracic aorta of rats in all Baihutang groups were significantly lower than those in model group(P<0.05). The mRNA level of fatty acid oxidation gene and the protein levels of IRS-1,PI3 K and Akt in liver were significantly increased(P<0.05),and the histopathology of thoracic aorta was significantly improved and the vascular wall thickness decreased significantly(P<0.05). Conclusion: Baihutang can reduce the levels of blood glucose,blood lipid and serum inflammatory factors in type 2 diabetic rats,regulate the expression of genes related to lipid metabolism in liver,and improve the histopathology and vascular remodeling of thoracic aorta. The mechanism may be related to the regulation of IRS-1/PI3 K/Akt signal pathway.
作者
郭杨志
杜娟
姜敏
GUO Yang-zhi;DU Juan;JIANG Min(Beijing Shijitan Hospital Affiliated to Capital Medical University,Beijing 100038,China;Changping District Integrated Chinese and Western Medicine Hospital of Beijing,Beijing 102208,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2021年第1期23-30,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81503503)。
作者简介
第一作者:郭杨志,博士,主治医师,从事中医药防治糖尿病、肿瘤及老年病研究.E-mail:p138990@163.com;通信作者:姜敏,主任医师,硕士生导师,从事中医药防治糖尿病、肿瘤及体制学研究,E-mail:jiangm1965@sina.com。
