TRPC6参与内质网应激诱导的肾小球系膜细胞凋亡  

TRPC6 involves in endoplasmic reticulum stress induced apoptosis of glomerular mesangial cells

作　　者：魏琳婷 葛蓬勃 马小琴[1] 高洁[1] 刘丹[1] 赵鹏[1] 魏士卓 董静 王莉[1] 付荣国[1] WEI Linting;GE Pengbo;MA Xiaoqin;GAO Jie;LIU Dan;ZHAO Peng;WEI Shizhuo;DONG Jing;WANG Li;FU Rongguo(Department of Nephrology,The Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004;Department of General Surgery,The First Affiliated Hospital of Xi’an Medical College,Xi’an 710077,China)

机构地区：[1]西安交通大学第二附属医院肾病科,陕西西安710004 [2]西安医学院第一附属医院普外科,陕西西安710077

出　　处：《西安交通大学学报（医学版）》2023年第6期898-903,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.82170697);陕西省自然科学基金资助项目(No.2022JM-472);中国医师协会医学循证专委会肾科学组PRO·润基金资助项目(KYJ202206-0003-3)。

摘　　要：目的探讨TRPC6离子通道在内质网应激(endoplasmic reticulum stress,ERS)诱导的肾小球系膜细胞(HBZY-1)凋亡中的作用及机制。方法实验分为正常对照组(NC)、thapsigargin(TG)、TG+SKF96365和TG+TRPC6siRNA组。qRT-PCR和Western blot技术检测TRPC6和ERS相关蛋白(GRP78和Caspase12)的mRNA和蛋白表达。流式细胞术和Hoechst33258方法检测细胞凋亡。Fluo-4AM Ca^(2+)成像技术测定细胞内Ca^(2+)内流(intracellular calcium,[Ca^(2+)]i)的变化情况。结果TG组细胞出现核浓缩或核碎裂为特征的凋亡形态变化,细胞凋亡率增加。TRPC6和ERS相关蛋白(GRP78和Caspase12)的表达明显高于NC组(P<0.05)。SKF96365和TRPC6 siRNA预孵育HBZY-1细胞可减轻细胞凋亡(P<0.05)。TG刺激后[Ca^(2+)]i也增加(P<0.05)。与TG组相比,SKF96365和TRPC6 siRNA处理后TRPC6、GRP78和Caspase12的表达下调,并伴有[Ca^(2+)]i的减少(P<0.05)。结论抑制TRPC6可以减轻TG诱导的HBZY-1细胞凋亡。Objective To explore the role and mechanism of TRPC6 in apoptosis of glomerular mesangial cells(HBZY-1)induced by endoplasmic reticulum stress(ERS).Methods The experiment groups were classified as follows:normal control(NC),thapsigargin(TG),TG+SKF96365,and TG+TRPC6 siRNA groups.Transcription and protein expressions of TRPC6 and ERS related proteins(GRP78 and Caspase12)were detected by qRT-PCR and Western blotting.Additionally,cell apoptosis was measured by flow cytometry and Hoechst33258.Finally,Fluo-4 AM Ca^(2+)imaging technique was used to determine changes of intracellular calcium([Ca^(2+)]i)by laser scanning confocal microscope.Results Morphological changes of apoptotic cells were characterized by nuclear enrichment or nuclear fragmentation,and the apoptosis rate was increased after TG stimulation.The expressions of TRPC6 and ERS related proteins(GRP78 and Caspase12)were elevated in TG group compared with NC group(P<0.05).Pre-incubation of HBZY-1 cells with SKF96365 and TRPC6 siRNA decreased cell apoptosis(P<0.05).The entry of[Ca^(2+)]i also increased after TG stimulation(P<0.05).The expressions of TRPC6,GRP78 and Caspase12 were downregulated compared with TG group after treatment with SKF96365 and TRPC6 siRNA accompanied by decreased[Ca^(2+)]i(P<0.05).Conclusion Taken together,this study suggests that inhibition of TRPC6 can alleviate TG-induced HBZY-1 cell apoptosis.

关 键 词：瞬时受体阳离子通道 毒胡萝卜素 肾小球系膜细胞 凋亡 肾脏纤维化 

分 类 号：R692[医药卫生—泌尿科学]