BK_(Ca)在高糖诱导的肾小球系膜细胞损伤中的作用及机制  

The role and mechanism of BK_(Ca) in mesangial cell injury induced by high glucose

作　　者：张雪 葛沛 顾立 李华 张丹 黄远琼 陈美娟 ZHANG Xue;GE Pei;GU Li;LI Hua;ZHANG Dan;HUANG Yuanqong;CHEN Meijuan(Department of Pharmacy,Southwest Medical University,Luzhou 646000,China;Department of Pharmacy,The Affiliated Hospital of Southwest Medical University,Luzhou 646000,China;Department of Nephrology,Luzhou Traditional Chinese Medicine Hospital,Luzhou 646000,China)

机构地区：[1]西南医科大学药学院,泸州646000 [2]西南医科大学附属医院药学部,泸州646000 [3]泸州市中医医院肾内科,泸州646000

出　　处：《西南医科大学学报》2024年第2期136-140,共5页Journal of Southwest Medical University

基　　金：四川省科技厅基金项目(2023NSFSC0666);泸州市科技局-西南医科大学联合基金项目(2019LZXNYDJ50);西南医科大学-泸州市中医医院基地项目(2018-LH005)。

摘　　要：目的探讨大电导钙激活钾通道(large conductance calcium-activated potassium channel,BK_(Ca))在糖尿病肾病中的作用及其机制。方法①用激动剂NS11021和抑制剂Tet及BK_(Ca)-siRNA干预后观察糖尿病肾病(diabetic nephropathy,DN)细胞模型的细胞增殖率、上清液中Ⅳ型胶原蛋白(collagenⅣ,ColⅣ)及纤连蛋白(fibronectin,FN)的表达变化,揭示BK_(Ca)的作用;②用激动剂NS11021和抑制剂Tet及BK_(Ca)-siRNA干预后观察DN细胞模型TGF-β_(1)、Smad2/3的含量变化;用激动剂TGF-β_(1)、抑制剂SB431542干预TGF-β_(1)/Smad2/3信号通路后,观察BK_(Ca)作用的变化,揭示BK_(Ca)作用与该信号通路相关性。结果①与高糖组比较,药理学激动BK_(Ca)使模型细胞BK_(Ca)β亚基蛋白表达增加、细胞增殖率增高,差异均具有统计学意义(P<0.05);与高糖组比较,药理学和基因抑制BK_(Ca),使其α、β亚基蛋白表达减少,从而出现模型细胞增殖率下降、ColⅣ和FN分泌明显减少(P<0.01)。②与高糖组比较,药理学激动BK_(Ca)引起Smad2/3含量增高,药理学和基因抑制BK_(Ca),引起TGF-β_(1)、Smad2/3含量明显降低,差异均具有统计学意义(P<0.05);与NS11021组比较,用SB431542阻断TGF-β_(1)/Smad2/3信号通路后,细胞增殖率明显降低,ColⅣ和FN分泌减少,差异均具有统计学意义(P<0.05)。结论高糖诱导的DN细胞模型存在BK_(Ca)高表达,抑制BK_(Ca)后细胞增殖率、ColⅣ和FN的分泌明显降低,从而改善DN病变,此作用与TGF-β_(1)/Smad2/3信号通路相关。Objective To explore the role and mechanism of BK_(Ca)(large conductance calcium-activated potassium channel,BK_(Ca))in DN(diabetic nephropathy,DN).Methods①After the intervention of agonist NS11021 and inhibitor Tet and BK_(Ca)-siRNA,the changes of cell proliferation rate,collagenⅣ(ColⅣ)and fibronectin(FN)in the DN cell model were observed to reveal the role of BK_(Ca);②After the intervention of agonist NS11021 and inhibitor Tet and BK_(Ca)-siRNA on BK_(Ca),the changes of TGF-β_(1)(Transforming growth factor beta 1)and Smad2/3 were observed in DN cell model;After the intervention of agonist TGF-β_(1) and inhibitor SB431542 on TGF-β_(1)/Smad2/3 signaling pathway,we observed the change of effects of BK_(Ca) on model cells,in order to reveal the correlation be-tween BK_(Ca) action and this signaling pathway.Results①Compared with high glucose group,pharmacologically excited BK_(Ca) increased the proliferation rate of model cells,and the expression of BK_(Ca)βsubunit protein was enhanced,the differences were statistically signifi-cant(P<0.05);compared with high glucose group,pharmacological and genetic inhibition of BK_(Ca) decreased the expression ofαandβsubunit protein,the proliferation rate of model cells decreased,and the secretion of ColⅣand FN decreased significantly(P<0.01).②Compared with the high glucose group,the content of Smad2/3 was increased due to the pharmacological excitation of BK_(Ca),the con-tent of TGF-β_(1) and Smad2/3 was significantly decreased due to the pharmacological and gene inhibition of BK_(Ca)(the differences were statistically significant(P<0.05);compared with NS11021 group,after blocking TGF-β_(1)/Smad2/3 signaling pathway with SB431542,the cell proliferation rate was significantly decreased,and the secretion of ColⅣand FN were decreased,and the differences were statis-tically significant(P<0.05).Conclusion BK_(Ca) was highly expressed in DN cell models induced by high glucose,and cell proliferation rate,ColⅣand FN secretion were significantly reduced after BK

关 键 词：糖尿病肾病 大电导钙激活钾通道 胶原Ⅳ 纤连蛋白 TGF-β_(1) SMAD2/3 

分 类 号：R96[医药卫生—药理学]