摘要
目的研究肾小管上皮细胞系HK-2低氧/复氧诱导的内质网应激(ERS)以及异氟醚对ERS诱导细胞凋亡的调控作用。方法将培养的HK-2肾小管上皮进行低氧/复氧处理,用2.56%异氟醚分别处理1、4和8 h后,检测乳酸脱氢酶(LDH)含量;用MTT法检测HK-2细胞增殖;用流式细胞仪检测细胞凋亡率;用RT-q PCR和半定量Western blot检测内质网应激标志蛋白GRP78、CHOP及内质网凋亡蛋白caspase-12。结果 2.56%异氟醚孵育4和8 h,LDH低于对照组,细胞增殖高于对照组(P<0.05);4和8 h时细胞凋亡/坏死率为24.2%±4.6%和13.3%±3.1%显著高于对照组的12.1%±1.3%和7.1%±1.1%(P<0.05)。结论异氟醚对低氧/复氧处理后促进的ERS及细胞凋亡具有抑制作用。
Objective To investigate the effects of isoflurane treatment on hypoxia/reoxygenation-induced endoplasmic reticulum stress（ ERS） and apoptosis in HK-2. Methods After treatment of hypoxia/reoxygenation in HK-2,2. 56%isoflurane was added and incubated for 1,4 and 8 h. Then the lactate dehydrogenase（ LDH） and MTT were measured to detect the hypoxia/reoxygenation-induced injury and cell viability in HK-2 cells. Flow cytometry was performed to detect apoptosis. RT-q PCR and semi-quantitative Western blot were employed for analyzing hallmarks of endoplasmic reticulum stress,GRP78,CHOP and caspase-12. Results After 4 and 8-hour co-incubation with 2. 56% isoflurane,LDH was lower and MTT was higher in treated groups,as compared with mock group（ P〈0. 05）,indicating the promoting effect of isoflurane treatment on cell survival. 1-hour co-incubation with 2. 56% isoflurane presented undetectable effect on cell survival. Consistently,Annexin V/PI double staining results present that,the apoptotic rates in 4 h-treated and 8 h-treated group were 12. 1%±1. 3% and 7. 1%±1. 1%,which were significantly lower than that in mock group 24. 2%±4. 6%and 13. 3%±3. 1%（ P〈0. 05）. GRP78 and caspase-12,but not CHOP were upregulated in treated cells as compared to untreated cells.After isoflurane treatment,caspase-12 activity was downregulated. Conclusions Isoflurane treatment inhibites ER stress-induced apoptosis in hypoxia/reoxygenation model cells by downregulating caspase-12 activity.
作者
唐静
胡玲
刘奉
李小山
赵梓亦
李国利
TANG Jing;HU Ling;LIU Feng;LI Xiao-shan;ZHAO Zi-yi;LI Guo-li(Dept.of Pathogen Biology and hnmunology;Pathology Teaching and Research Office,Dept.of Basic Medicine;Dept.of Clinical Medicine,Chongqing Three Gorges Medical College,Chongqing 404120;the Affiliated Hospital of Chengdn University of Traditional Chinese Medicine,Chengdu 641000,China)
出处
《基础医学与临床》
CSCD
2018年第9期1258-1262,共5页
Basic and Clinical Medicine
基金
重庆三峡医药高等专科学校校级苗圃工程(2016mpxz19)
重庆市卫计委中医项目(zy201602100)
四川省科技支撑项目(2016FZ0093)
关键词
异氟醚
内质网应激
低氧/复氧模型
细胞凋亡
isoflurane
endoplasmic reticulum stress（ER stress）
hypoxia/reoxygenation
apoptosis
