摘要
为探讨环状RNA VMA21(circular RNA VMA21,circVMA21)对高糖诱导的肾小管上皮细胞损伤的影响及分子机制,用25 mmol/L葡萄糖处理肾小管上皮细胞株HK-2作为高葡萄糖(high glucose,HG)组,以5.5 mmol/L葡萄糖处理的细胞作为对照组。用qRT-PCR检测circVMA21和miR-665的表达水平;ELISA检测IL-6、TNF-α水平;FACS检测细胞凋亡情况;Western blotting检测蛋白表达水平;双荧光素酶报告基因试验检测circVMA21和miR-665的靶向关系。结果显示,高糖诱导的肾小管上皮细胞中circVMA21表达水平降低,miR-665表达水平升高,IL-6、TNF-α水平升高,细胞凋亡率升高,B淋巴细胞瘤2(B-cell lymphoma 2,Bcl-2)表达水平降低,Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)表达水平升高(P<0.05)。circVMA21过表达或抑制miR-665表达后,高糖诱导的肾小管上皮细胞中IL-6、TNF-α水平降低,细胞凋亡率降低,Bcl-2表达水平升高,Bax表达水平降低(P<0.05)。circVMA21可靶向调控miR-665;miR-665过表达逆转了circVMA21过表达对高糖诱导的肾小管上皮细胞的作用。该研究提示,circVMA21过表达可通过靶向下调miR-665保护肾小管上皮细胞免受高糖诱导的损伤。
This study aims to investigate the effect of circular RNA VMA21(circVMA21)on renal tubular epithelial cell damage induced by high glucose and the underlying molecular mechanism.To this end,renal tubular epithelial cell line HK-2 was treated with 25 mmol/L glucose(used as the high glucose[HG]group)or 5.5 mmol/L glucose(used as the control group).qRT-PCR was performed to detect the expression levels of circVMA21 and miR-665.ELISA was used to detect the levels of IL-6 and TNF-α.FACS was used to detect cell apoptosis.Western blotting was used to detect related protein expression.A dual luciferase reporter experiment was used to detect the interaction between circVMA21 and miR-665.The results showed that in the HG group,the expression level of circVMA21 in renal tubular epithelial cells was decreased,and the expression level of miR-665 and the protein levels of IL-6,and TNF-αwere increased,accompanied by increased apoptosis rate and B-cell lymphoma 2(Bcl-2)expression and decreased Bcl-2 associated X protein(Bax)expression(P<0.05).In contrast,overexpression of circVMA21 or inhibition of miR-665 caused decreased levels of IL-6 and TNF-αand apoptosis rate as well as increased Bcl-2 and decreased Bax protein levels in HG group renal tubular epithelial cells(P<0.05).circVMA21 targeted and regulated miR-665 and miR-665 overexpression reversed the effect of circVMA21 overexpression on renal tubular epithelial cells induced by high glucose.This study suggests that overexpression of circVMA21 may protect renal tubular epithelial cells from high glucose-induced damage by down-regulation of miR-665.
作者
黄成丽
李根
林芳荣
HUANG Cheng-li;LI Gen;LIN Fang-rong(Department of Nephrology,the First Affiliated Hospital of Hainan Medical College,Haikou 570145,China)
出处
《现代免疫学》
CAS
北大核心
2023年第1期48-53,共6页
Current Immunology
