| 常杰.微小 RNA?204?3p通过靶向调控 Krüppel样因子 6对高糖诱导小鼠足细胞损伤的保护作用[J].安徽医药,2020,24(12):2433-2439. |
| 微小 RNA?204?3p通过靶向调控 Krüppel样因子 6对高糖诱导小鼠足细胞损伤的保护作用 |
| Study on the protective mechanism of overexpression of microRNA?204?3p on mouse podocytes injury induced by high glucose through targeting Krüppel?like factor 6 |
| |
| DOI：10.3969/j.issn.1009?6469.2020.12.026 |
| 中文关键词: 锌指蛋白 结蛋白 bcl?2相关 X蛋白质 微小 RNA?204?3p Krüppel样因子 6 高糖 足细胞 小鼠 |
| 英文关键词: Zinc finger protein Desmin bcl?2?associated X protein miR?204?3p Krüppel?like factor 6 High glucose Podocyte Mice |
| 基金项目: |
|
| 摘要点击次数: 1547 |
| 全文下载次数: 486 |
| 中文摘要: |
| 目的探讨微小 RNA?204?3p(miR?204?3p)对高糖诱导小鼠足细胞 MPC5损伤的保护作用及其分子机制。方法采用高糖(30 mmol/L D?葡萄糖)处理 MPC5细胞。实时定量 PCR(qPCR)检测细胞中 miR?204?3p与锌指蛋白 Krüppel样因子 6(KLF6) mRNA水平。 MPC5细胞分为 NG组(正常对照)HG组(高糖刺激)HG+miR?204?3p组、 HG+miR?NC组、 HG+si?KLF6组、 HG+si? NC组， HG+miR?204?3p+pcDNA?KLF6组和HG+m，iR?204?3p+pcDNA组，。蛋白质印迹法(Western Blot)检测肌动蛋白微丝偶联蛋白(synaptopodin)、结蛋白(desmin)、 B细胞淋巴瘤 /白血病 ?2(Bcl?2)、 Bcl?2相关 X蛋白(Bax)与活化的含半胱氨酸的天冬氨酸蛋白水解酶 3(cleaved?caspase?3)蛋白表达，流式细胞仪检测细胞凋亡，生物学信息预测和双荧光素酶报告基因分析 miR?204? 3p和 KLF6的靶向关系。结果与正常对照相比，高糖可显著降低 MPC5细胞中 miR?204?3p表达量［(0.41±0.04)比(1.02± 0.08)］上调 KLF6 mRNA［(2.86±0.27)比(1.03±0.08)］和蛋白［(0.89±0.08)比(0.46±0.04)］水平，还可上调 KLF6、desmin［(0.81± 0.07)比(，0.34±0.03)］、 Bax、cleaved?caspase?3蛋白表达及细胞凋亡率［(22.46±2.08)%比(5.26±0.64)%］下调 synaptopodin蛋白［(0.34±0.03)比(0.76±0.07)］及 Bcl?2蛋白表达量(P＜0.05)。与 HG+miR?NC组比较， HG+miR?204?3p组明，显提高 synaptopodin［(0.67±0.06)比(0.32±0.03)］及 Bcl?2蛋白水平，降低 desmin［(0.41±0.04)比(0.83±0.08)］、 Bax、cleaved?caspase?3蛋白水平及细胞凋亡率［(11.25±1.65)%比(24.58±2.47)%］(P＜0.05)。与 HG+si?NC组比较， HG+si?KLF6组明显提高 synaptopodin［(0.62±0.06)比(0.28±0.03)］及 Bcl?2蛋白水平，降低 desmin［(0.49±0.05)比(0.86±0.08)］、 Bax蛋白水平及细胞凋亡率［(14.69±1.87)%比(25.47±2.68)%］(P＜0.05)。 miR?204?3p直接靶向 KLF6。与 HG+miR?204?3p+pcDNA组比较， HG+miR?204?3p+pcDNA?KLF6组显著增加高糖刺激小鼠肾脏足 MPC5细胞中 KLF6、desmin［(0.68±0.06)比(0.36±0.04)］、 Bax蛋白表达量和细胞凋亡率［(18.41±1.67)%比(11.05±1.02)%］，显著减少 synaptopodin蛋白［(0.38±0.03)比(0.69±0.07)］和 Bcl?2蛋白表达量(P＜0.05)。结论 miR?204?3p通过直接靶向 KLF6抑制高糖刺激的足细胞凋亡，保护细胞损伤。 |
| 英文摘要: |
| Objective To investigate the protective effects of microRNA?204?3p(miR?204?3p)on high glucose?induced mouse podocytes(MPC5)injury and its molecular mechanism.Methods MPC5 cells were treated with high glucose(30 mmol/L D?glu? cose).The levels of miR?204?3p and Krüppel?like factor 6(KLF6)mRNA in MPC5 cells were detected by real?time quantitative PCR(qPCR).MPC5 cells were assigned into NG group(normal control)，HG group(high glucose stimulation)，HG+miR?204?3p group，HG+miR?NC group，HG+si?KLF6 group，HG+si?NC group，HG+miR?204?3p+pcDNA?KLF6 group and HG+miR?204?3p+pcD? NA group.Western Blot was used to detect synaptopodin，desmin，B cell lymphoma/leukemia?2(Bcl?2)，Bcl?2 related X protein(Bax)and activated cysteine?containing aspartate proteolytic enzyme?3(cleaved?caspase?3)protein expression.Flow cytometry was used to detect apoptosis，biological information prediction and double luciferase reporter gene analysis the targeting relationship of miR?204?3p and KLF6.Results Compared with normal controls，high glucose significantly reduced the expression of miR?204?3p in MPC5 cells［(0.41±0.04)vs.(1.02±0.08)］up?regulated KLF6 mRNA［(2.86±0.27)vs.(1.03±0.08)］ and protein［(0.89±0.08)vs.(0.46±0.04)］levels，up?regulated KLF6esmin［(0.81±0.07)vs.(0.34±0.03)］，Bax，cleaved?caspase?3 protein expres? sion and cell apoptosis death rate［(22.46±2.08)% vs.(5.26±0.64)%］，and down?regulated synaptopodin protein［(0.34±0.03)vs.，d，(0.76±0.07)］ and Bcl?2 protein expression(P＜0.05).Compared with the HG+miR?NC group，the HG+miR?204?3p group signifi?cantly increased synaptopodin［(0.67±0.06)vs.(0.32±0.03)］ and Bcl?2 protein levels，and decreased desmin［(0.41±0.04)vs.(0.83±0.08)］，Bax，cleaved?caspase?3 protein levels and apoptosis rate［(11.25±1.65)% vs.(24.58±2.47)%］(P＜0.05).Compared with the HG+si?NC group，the HG+si?KLF6 group significantly increased synaptopodin［(0.62±0.06)vs.(0.28±0.03)］ and Bcl?2 protein levels，and decreased desmin［(0.49±0.05)vs.(0.86±0.08)］，Bax protein level and apoptosis rate［(14.69±1.87)% vs.(25.47±2.68)%］(P＜0.05).miR?204?3p directly targets KLF6.Compared with the HG+miR?204?3p+pcDNA group，the HG+miR? 204?3p+pcDNA?KLF6 group significantly increased the ratio of KLF6 and desmin［(0.68±0.06)vs.(0.36±0.04)］，Bax protein ex? pression and cell apoptosis rate［(18.41±1.67)% vs.(11.05±1.02)%］in the MPC5 cells，and significantly reduced synaptopodin protein［(0.38±0.03)vs.(0.69±0.07)］ and Bcl?2 Protein expression(P＜0.05).Conclusion miR?204?3p directly targets SGK1 to inhibit apoptosis of podocytes stimulated by high glucose and protects cell damage. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
