瞬间受体电位通道蛋白活化可减轻丙泊酚引起的SH-SY5Y细胞毒性  

The activation of TRPC channel can alleviate propofol-induced cytotoxicty of SH-SY5Y cells

作　　者：黄斯伦 佘应军[1] 张锐忠[1] 宋兴荣[1] 夏慧敏[1] 

机构地区：[1]广州医科大学附属广州市妇女儿童医疗中心,510623

出　　处：《临床麻醉学杂志》2015年第10期1007-1010,共4页Journal of Clinical Anesthesiology

基　　金：国家临床重点专科建设项目(小儿外科)(5001-2140019);广州市医药卫生科技项目(20131A011050)

摘　　要：目的探讨经典瞬间受体电位通道蛋白（transient receptor potential-canonical channel,TRPC）在丙泊酚诱导SH-SY5Y细胞毒性中的作用。方法实验一：细胞随机分为七组（每组设5个平行副孔）：对照组（C组）正常培养,丙泊酚溶剂组（A组）加入20%脂肪乳剂,不同浓度丙泊酚组（P1~P5组）终浓度分别为5、10、50、100、200μM,各组分别孵育SH-SY5Y细胞72h;200μM丙泊酚处理SH-SY5Y细胞不同时间（12、24、36、48、72h）,四唑单钠盐-WST-8（Cell Counting Kit-8,CCK-8）测定细胞存活率。实验二：SH-SY5Y细胞随机分为四组：丙泊酚溶剂组（A组）加入20%脂肪乳剂;P组：200μM丙泊酚;T1组：200μM丙泊酚＋TRPC通道激动剂OAG（25μM）;T2组：200μM丙泊酚＋TRPC通道拮抗剂SKF96365（3μM）。细胞处理72h后,CCK-8监测各组细胞存活率,AnnexinV-FLUOS流式细胞术检测细胞凋亡。结果实验一：C组与A组SH-SY5Y细胞存活率差异无统计学意义;P1~P3组细胞存活率上升,P4、P5组细胞存活率降低（P〈0.05）。实验二：细胞孵育72h后,P组SH-SY5Y细胞存活率为60.2%,凋亡率为18.5%±1.7%,C组存活率为100%,凋亡率为1.8%±0.8%（P〈0.05）;与P组比较,T1组细胞存活率为86.4%,上升了26.2%（P〈0.01）,凋亡率为4.0%±0.9%（P〈0.05）;T2组存活率为49.2%,较P组降低了11.0%（P〈0.05）,凋亡率为（34.1±2.6）%（P〈0.01）。结论 TRPC通道活化可减轻200μM丙泊酚诱导下的SH-SY5Y细胞毒性作用。Objective To study the effect of transient receptor potential-canonical channel（TRPC）on propofol-induced cytotoxicty of SH-SY5 Ycells.Methods Test 1：The cells were randomly divided into 7groups（n=5）：control group（group C）received no treatment,in group A 20%lipid emulsion was added,in group P1-P5 propofol was added at the final concentration of 5,10,50,100,200μM respectively,each group of SH-SY5 Y were incubated with propofol for 72 h,and the cells were treated with 200μM propofol for different time course（12,24,36,48,72h）,the cell survival rate of each group was detected by tetrazolium salts-WST-8（Cell Counting Kit-8,CCK-8）.Test 2：The SH-SY5 Ycells were randomly divided into 4groups：group A was added with 20% emulsion,group P with 200μM propofol,group T1 with TRPC excitomotor（OAG 25μM）and group T2 with TRPC antagonist（SKF96365 3μM）.After treatment of 72 h,each group was detected by CCK-8to observe the cell survival rate and by Annexin V to survey the cell’s apoptosis.Results Test 1：There was no significantly difference in the cell survival rate between group C and A;Propofol at the dose of5-50μM in P1-P3 could improve cell survival rate,but at higher doses in P4-P5（100-200μM）,propofol appears to be cytotoxic.Test 2：After 72 hincubation,compared with group C（cell viability is 100% and apoptosis rate is 1.8% ±0.8%）,there was significantly difference in the cell viability（60.2%）and the apoptosis rate 18.5%±1.7%in group P;the cell viability of group T1 was 86.4%,26.2% higher than in group P（P〈0.01）,and the apoptosis rate was 4.0% ±0.9%（P〈0.05 vs group P）;the cell viability of group T2 was 49.2%,11.0%lower than in group P（P〈0.05）,and the apoptosis rate of group T2 was 34.1%±2.6%（P〈0.01）.Treatment with TRPC channel blocker or activator alone did not obviously affect the survival rate of the cells.Conclusion The activation of TRPC channel can alleviate 200μM propofol-induced cytotoxicty of SH-SY5 Ycells.

关 键 词：丙泊酚 SH-SY5Y细胞 TRPC通道 细胞存活率 凋亡 

分 类 号：R726.1[医药卫生—儿科]