摘要
目的探讨槲皮素对人肾小管上皮HK-2细胞脂质代谢的影响及其机制。方法采用25.0 mmol·L-1葡萄糖诱导HK-2细胞建立高糖模型,在CCK-8检测槲皮素对HK-2细胞活性影响的基础上,采用油性红O染色观察细胞内脂质沉积;化学酶法测定细胞内胆固醇含量;Real-time PCR和Western blot分别检测低密度脂蛋白受体(LDLr)、固醇调节元件结合蛋白裂解激活蛋白(SCAP)、胆固醇调节元件结合蛋白2(SREBP-2)和羟甲基戊二酸单酰辅酶A还原酶(HMGCR)mRNA转录和蛋白表达水平。结果与对照组相比,高糖模型组不仅HK-2细胞内脂滴积分光密度(IOD)值显著增加而且含脂滴的细胞数明显增多(P<0.05),干预48 h后高糖槲皮素组HK-2细胞内IOD值较高糖模型组明显下降,含脂滴的细胞数也明显减少(P<0.05);高糖模型组HK-2细胞内总胆固醇和游离胆固醇含量较对照组明显升高(P<0.05),干预48 h后高糖槲皮素组HK-2细胞内总胆固醇和游离胆固醇含量较高糖模型组显著下降(P<0.05);高糖模型组HK-2细胞内LDLr、SCAP、SREBP-2和HMGCR mRNA和蛋白表达量均较对照组显著升高(P<0.05),干预48 h后高糖槲皮素组HK-2细胞LDLr、SCAP、SREBP-2和HMGCR mRNA转录和蛋白表达量均较高糖模型组显著降低(P<0.05)。结论槲皮素可减少肾小管上皮细胞内脂质沉积和降低细胞内胆固醇含量,其机制可能与抑制SCAP-SREBP2-LDLr通路有关。
Objective To investigate the effect of quercetin on lipid metabolism and its underlying mechanism in human renal tubular epithelial HK-2 cells.Methods HK-2 cells were induced by 25 mmol·L-1 glucose to establish a high glucose model.Based on the activity of quercetin on HK-2 cells detected by CCK-8 assay,lipid deposition was observed after oil red O staining and intracellular cholesterol esters were determined by chemical enzymatic method in HK-2 cells.Furthermore,Real-time PCR and Western blot assay were used to detect the mRNA and protein expression levels of LDLr,SCAP,SREBP-2 and HMGCR.Results Compared with control group,the integral optical density(IOD)of lipid droplets in HK-2 cells and the number of cells containing lipid droplets in high glucose model group increased significantly.However,the IOD of lipid droplets in HK-2 cells and the number of cells containing lipid droplets decreased remarkably after treatment with quercetin for 48 h(P<0.05).Compared with control group,the total and free cholesterol contents in cells from high glucose model group were remarkably higher(P<0.05),which decreased markedly after treatment with quercetin for 48 h(P<0.05).Similarly,the expressions of LDLr,SCAP,SREBP-2 and HMGCR mRNA and protein were markedly higher in the high glucose model group(P<0.05),which were down-regulated after treatment with quercetin for 48 h(P<0.05).Conclusion Quercetin can reduce lipid deposition and decrease cholesterol level in renal tubular epithelial cells,which may be related to the inhibition of SCAP-SREBP2-LDLr Pathway.
作者
蒋曦媛
韩筱清
李楠
余江毅
JIANG Xiyuan;HAN Xiaoqing;LI Nan;YU Jiangyi(Kunshan Hospital Affiliated to Nanjing University of Traditional Chinese Medicine,Kunshan 215300,China;First Clinical Medical College of Nanjing University of Traditional Chinese Medicine,Nanjing 210029,China)
出处
《沈阳药科大学学报》
CAS
CSCD
北大核心
2019年第12期1107-1112,共6页
Journal of Shenyang Pharmaceutical University
基金
江苏省中医药管理局一般项目(YB2015104).
作者简介
蒋曦媛(1980-),女(汉族),江苏南京人,副主任医师,硕士研究生,主要从事糖尿病慢性并发症的防治研究,Tel.13962698009,E-mail:397365264@qq.com。
