摘要
目的:探讨银杏内酯B(GB)对高糖(HG)诱导的肾小管上皮细胞凋亡及炎症因子的影响及其可能作用机制。方法:采用HG诱导人肾小管上皮细胞HK-2建立细胞损伤模型,不同剂量的GB处理HK-2细胞,并将anti-miR-NC、anti-miR-155-5p分别转染至HK-2细胞后加入25 mmol/L葡萄糖处理24 h,将miR-NC、miR-155-5p mimics分别转染至HK-2细胞后加入100μmol/L GB与25 mmol/L葡萄糖共同处理24 h;采用流式细胞术检测细胞凋亡率;ELISA法检测炎症因子IL-6、TNF-α的水平;RT-qPCR法检测miR-155-5p的表达量;Western blot检测Bcl-2相关X蛋白(Bax)、B淋巴细胞瘤(Bcl-2)蛋白表达水平。结果:GB处理后,高糖诱导的HK-2细胞凋亡率、IL-6、TNF-α水平、miR-155-5p表达量和Bax蛋白表达水平均降低(P<0.05),Bcl-2蛋白表达升高(P<0.05),且呈剂量依赖性;转染anti-miR-155-5p后,高糖诱导的HK-2细胞凋亡率、IL-6、TNF-α水平、miR-155-5p表达量和Bax蛋白表达水平降低(P<0.05),Bcl-2蛋白表达水平升高(P<0.05);转染miR-155-5p mimics可减弱GB对HG诱导的HK-2细胞凋亡及炎症因子表达的影响。结论:GB可通过抑制细胞凋亡及炎症因子表达来减轻HG诱导的肾小管上皮细胞损伤,其作用机制可能与抑制miR-155-5p表达有关。
Objective:To explore the effect of ginkgolide B(GB)on renal tubular epithelial cell apoptosis and inflammatory factors induced by high glucose(HG)and its possible mechanism.Methods:Human renal tubular epithelial cells HK-2 were induced by HG to establish a cell injury model.HK-2 cells were treated with different doses of GB,and anti-miR-NC and anti-miR-155-5p were transfected into HK-2 cells,respectively,and then treated with 25 mmol/L glucose for 24 h,miR-NC and miR-155-5p mimics were transfected into HK-2 cells and treated with 100μmol/L GB and 25 mmol/L glucose for 24 h.Flow cytometry was used to detect the rate of apoptosis,ELISA was used to detect the levels of IL-6 and TNF-α,qRT-PCR was used to detect the expression of miR-155-5p,the expressions of Bcl-2 associated X protein(Bax)and B lymphoblastoma(Bcl-2)were detected by Western blot.Results:After GB treatment,the apoptosis rate,IL-6,TNF-α,miR-155-5p expression and Bax protein expression level of HK-2 cells induced by high glucose were decreased(P<0.05),and the expression level of Bcl-2 protein was increased(P<0.05)in a dose-dependent manner.After transfection with anti-miR-155-5p,the apoptosis rate,IL-6,TNF-α,miR-155-5p expression and Bax protein expression level of HK-2 cells induced by high glucose were decreased(P<0.05),and the expression level of Bcl-2 protein was increased(P<0.05).Transfection of miR-155-5p mimics could attenuate the effect of ginkgolide B on HK-2 cell apoptosis and the expression of inflammatory factors induced by HG.Conclusion:GB can alleviate HG-induced renal tubular epithelial cell injury by inhibiting apoptosis and expression of inflammatory factors,the mechanism of which may be related to inhibiting the expression of miR-155-5p.
作者
秦美灵
吴春丽
陈延玲
廉波
尚海玉
韩英
QIN Mei-ling;WU Chun-li;CHEN Yan-ling;LIAN Bo;SHANG Hai-yu;HAN Ying(Department of Endocrinology,Jinan Hospital of Integrated Chinese and Western Medicine,Jinan 271100,Shandong,China;Treatment and Prevention Center,Jinan Hospital of Integrated Chinese and Western Medicine,Jinan 271100,Shandong,China;Department of Finance,Jinan Hospital of Integrated Chinese and Western Medicine,Jinan 271100,Shandong,China)
出处
《川北医学院学报》
CAS
2024年第2期150-155,共6页
Journal of North Sichuan Medical College
基金
山东省济南市科技计划项目(202134066)。
