左卡尼汀抑制高糖诱导的肾小球系膜细胞增殖并诱导自噬和凋亡的研究  

L-carnitine inhibited the proliferation and induced autophagy and apoptosis of mesangial cells induced by high glucose

作　　者：柴海霞 刘静梅 王会军 王印华 郭炜妍 CHAI Hai-xia;LIU Jing-mei;WANG Hui-jun;WANG Yin-hua;GUO Wei-yan(Department of Critical Care Medicine,Affiliated Hospital of North China University of Technology,Tangshan 063000,Hebei Province,China)

机构地区：[1]华北理工大学附属医院重症医学科,河北唐山063000

出　　处：《中国临床药理学杂志》2023年第11期1577-1581,共5页The Chinese Journal of Clinical Pharmacology

基　　金：河北省医学科学研究课题计划基金资助项目(20211702)。

摘　　要：目的研究左卡尼汀对高糖诱导的肾小球系膜细胞自噬、凋亡和增殖的影响机制。方法将HMC细胞随机分为NC、Man、HG、LC、sh-TRPC6及sh-TRPC6+AICAR组。NC组、Man组及HG组中HMC细胞分别用正常糖培养基、高渗培养基及高糖培养基培养。将HG组HMC细胞经1.8 mmol·L^(-1)左卡尼汀处理后,随机分为LC组、sh-TRPC6组及sh-TRPC6+AICAR组。sh-TRPC6组及sh-TRPC6+AICAR组转染sh-TRPC6,sh-TRPC6+AICAR组再经AICAR处理。以蛋白质印迹法检测各组HMC细胞中瞬时受体电位阳离子通道6(TRPC6)、腺苷一磷酸活化蛋白激酶/西罗莫司靶蛋白信号通路(AMPK/mTOR)通路相关蛋白及自噬相关蛋白的表达水平,以细胞计数法-8(CCK-8)检测各组HMC细胞的增殖活力,以Annexin V-FITC/PI检测各组HMC细胞的凋亡水平。结果在高糖条件下,HMC细胞增殖活力显著上调,且自噬和凋亡水平被抑制(P<0.05)。LC组和HG组的增殖活力(光密度值)分别为0.65±0.04和0.86±0.05,轻链3B(LC3B)Ⅰ蛋白相对表达水平分别为0.26±0.04和1.00±0.05,LC3BⅡ蛋白相对表达水平分别为1.46±0.04和1.01±0.07,凋亡率分别为(12.53±0.50)%和(3.90±0.40)%,差异均有统计学意义(均P<0.05)。左卡尼汀能够抑制高糖诱导的HMC细胞增殖,并促进其自噬和凋亡。在高糖条件下,HMC细胞中TRPC6表达水平和AMPK/mTOR信号通路被抑制,且左卡尼汀能够上调HMC细胞中TRPC6的表达水平和激活AMPK/mTOR信号通路(均P<0.05)。结论左卡尼汀通过上调TRPC6激活AMPK/mTOR信号通路,从而抑制高糖诱导的HMC细胞增殖,并诱导其自噬和凋亡。Objective To explore the mechanism of L-carnitine on autophagy,apoptosis and proliferation of glomerular mesangial cells induced by high glucose.Methods HMC cells were randomly divided into NC,Man,HG,LC,sh-TRPC6 and sh-TRPC6+AICAR groups.Normal glucose medium,hypertonic medium and high glucose medium were used for HMC cells in NC group,Man group and HG group,respectively.The HMC cells in the HG group were treated with 1.8 mmol·L^(-1) L-carnitine and then randomly divided into the LC group,the sh-TRPC6 group and the sh-TRPC6+AICAR group.The sh-TRPC6 group and the sh-TRPC6+AICAR group were transfected with sh-TRPC6.In addition,the sh-TRPC6+AICAR group was treated with AICAR.Western blotting was used to detect the expression levels of canonical transient receptor potential 6(TRPC6),adenosine monophosphate activated protein kinase/mammalian target of rapamycin(AMPK/mTOR)pathway-related proteins and autophagy-related proteins in HMC cells of each group.Cell counting kit-8(CCK-8)was used to detect the proliferation activity of HMC cells in each group.Annexin V-FITC/PI was used to detecte the apoptosis level of HMC cells in each group.Results Under high glucose conditions,the proliferation activity of HMC cells was significantly increased,and the levels of autophagy and apoptosis were inhibited(P<0.05).The proliferative activities(optical density)of LC group and HG group were 0.65±0.04 and 0.86±0.05;the expression levels of light chain 3B(LC3B)Ⅰprotein were 0.26±0.04 and 1.00±0.05;the expression levels of LC3BⅡprotein were 1.46±0.04 and 1.01±0.07;the apoptosis rates were(12.53±0.50)%and(3.90±0.40)%;the differences were statistically significant(all P<0.05).L-carnitine inhibited the proliferation of HMC cells induced by high glucose and promoted autophagy and apoptosis.Under high glucose conditions,the expression level of TRPC6 and AMPK/mTOR signaling pathway in HMC cells were inhibited,and L-carnitine upregulated the expression level of TRPC6 in HMC cells and activated AMPK/mTOR signaling pathway(all P<0.05

关 键 词：左卡尼汀 瞬时受体电位阳离子通道6 腺苷一磷酸活化蛋白激酶/西罗莫司靶蛋白信号通路 肾小球系膜细胞 

分 类 号：R97[医药卫生—药品]