葛根素调控miR-30e-5p缓解高糖所致的足细胞损伤  

Puerarin Regulating miR-30e-5p to Alleviate High Glucose-Induced Podocyte Injury

作　　者：梅兰 郭阳 张宁 傅盼盼 孙国钧[2] MEI Lan;GUO Yang;ZHANG Ning;FU Panpan;SUN Guojun(Heping District,General Hospital of Northern Theater Command,Shenyang Liaoning China 110000;Dalian Friendship Hospital,Dalian Liaoning China 116000)

机构地区：[1]北部战区总医院和平院区,辽宁沈阳110000 [2]大连市友谊医院,辽宁大连116000

出　　处：《中医学报》2022年第10期2184-2190,共7页Acta Chinese Medicine

基　　金：大连市卫生局科研基金项目(2016-0258X)。

摘　　要：目的:探讨葛根素能否通过调控miR-30e-5p的表达减轻高糖所致的足细胞损伤。方法:高糖诱导小鼠肾足细胞MPC5建立足细胞损伤模型,使用不同剂量的葛根素处理细胞;将miR-NC、miR-30e-5p mimics、anti-miR-NC、anti-miR-30e-5p转染至MPC5细胞后采用高糖诱导建立足细胞损伤模型,并给予不同剂量的葛根素处理细胞。采用流式细胞术检测细胞凋亡率;采用RT-qPCR法检测miR-30e-5p的表达量;Western Blot法检测半胱氨酰天冬氨酸特异性蛋白酶3(cysteinyl aspartate-specific protease-3,caspase-3)、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bcl-2-associated X protein,Bax)、p62、微管相关蛋白轻链3Ⅱ(microtubule-associated protein l light chain 3,LC3-Ⅱ)、LC3-Ⅰ蛋白表达水平并计算LC3-Ⅱ/LC3-Ⅰ的比值。结果:高糖处理小鼠肾足细胞MPC5后,可明显提高MPC5细胞的凋亡率及caspase-3、Bax、p62蛋白表达水平(P<0.05),显著降低Bcl-2蛋白及miR-30e-5p的表达水平和LC3-Ⅱ/LC3-Ⅰ的比值(P<0.05),而葛根素可显著降低细胞凋亡率及caspase-3、Bax、p62的蛋白表达水平(P<0.05),显著升高Bcl-2蛋白、miR-30e-5p的表达水平及LC3-Ⅱ/LC3-Ⅰ的比值(P<0.05);与miR-NC组比较,miR-30e-5p组细胞凋亡率显著降低(P<0.05),caspase-3、Bax、p62蛋白表达水平显著降低(P<0.05),Bcl-2蛋白水平、LC3-Ⅱ/LC3-Ⅰ的比值显著升高(P<0.05);与高剂量葛根素+anti-miR-NC组比较,高剂量葛根素+anti-miR-30e-5p组细胞凋亡率显著升高(P<0.05),caspase-3、Bax、p62蛋白表达水平显著升高(P<0.05),Bcl-2蛋白表达水平显著降低(P<0.05),LC3-Ⅱ/LC3-Ⅰ的比值显著降低(P<0.05)。结论:葛根素可通过上调miR-30e-5p的表达抑制高糖诱导的足细胞凋亡,其作用机制可能与诱导细胞自噬有关。Objective: To investigate whether puerarin can alleviate high glucose-induced podocyte injury by regulating the expression of miR-30e-5p.Methods: High glucose induced mouse renal podocyte MPC5 cells to establish a podocyte injury model, and treated the cells with different doses of puerarin;miR-NC,miR-30e-5p mimics,anti-miR-NC,anti-miR-30e MPC5 cells were transfected with-5p to establish a podocyte injury model induced by high glucose, and the cells were treated with different doses of puerarin.The apoptosis rate was detected by flow cytometry;the expression of miR-30e-5p was detected by qRT-PCR;the cysteyl aspartate-specific protease-3(cysteinyl aspartate-specific protease-3) was detected by Western blot.,Caspase-3),Bcl-2-associated X protein(Bax),B-cell lymphoma-2(Bcl-2),p62,microtubule-associated Protein light chain 3 Ⅱ(microtubule-associated protein l light chain 3,LC3-Ⅱ),microtubule-associated protein l light chain 3(LC3-Ⅰ) protein expression and calculation of LC3-Ⅱ/LC3-Ⅰ ratio.Results: After high glucose treatment of MPC5 in mouse renal podocytes, the apoptosis rate and the protein levels of caspase-3,Bax and p62 in MPC5 cells were significantly increased(P<0.05),and the protein levels of Bcl-2 and miR-30e-5p were significantly decreased.The expression of LC3-Ⅱ/LC3-Ⅰ and the ratio of LC3-Ⅱ/LC3-Ⅰ(P<0.05),while puerarin can significantly reduce the apoptosis rate and the protein levels of caspase-3,Bax and p62(P<0.05),and significantly increase Bcl-2 The expression of protein, miR-30e-5p and LC3-Ⅱ/LC3-Ⅰ ratio(P<0.05);compared with miR-NC group, the apoptosis rate of miR-30e-5p group was significantly decreased(P<0.05),caspase-3,Bax, p62 protein levels were significantly decreased(P<0.05),Bcl-2 protein levels, LC3-Ⅱ/LC3-Ⅰ ratio were significantly increased(P<0.05);compared with high-dose puerarin+anti-miR-NC Compared with other groups, the apoptosis rate of high-dose puerarin+anti-miR-30e-5p group was significantly increased(P<0.05),the protein levels of caspase-3,Bax, p62 were signi

关 键 词：葛根素 miR-30e-5p 高糖 足细胞 细胞凋亡 细胞自噬 糖尿病肾病 

分 类 号：R285.5[医药卫生—中药学]