摘要
目的观察内质网应激(Endoplasmic ReticulumStress,ERS)对自身免疫性脑炎(Autoimmune Encephalitis,AE)病人T淋巴细胞存活和凋亡的影响以及未折叠蛋白反应(Unfolded Protein Response,UPR)信号通路在AE淋巴细胞存活和死亡中的作用。方法本研究在2017年2月至2019年2月选取石家庄市第一医院儿科AE病人和健康供者外周血样本进行研究,并用不同浓度的毒胡萝卜素(Thapsigargin,TG)(浓度为0、0.5、1、5、10 nM)分别处理AE病人和健康供者的T淋巴细胞48 h,然后利用八肽胆囊收缩素(cholecystokinin octapeptide,CCK-8)实验方法检测细胞存活能力;用TG(浓度为5 nM)分别处理AE病人和健康供者的T淋巴细胞48 h,流式细胞仪检测TG未处理以及TG处理的T淋巴细胞的细胞凋亡情况;TG(浓度为5 n M)刺激AE病人和健康供者的T淋巴细胞48 h,蛋白质印迹法(Western blot)检测UPR标志分子免疫球蛋白结合蛋白(BIP)和同源蛋白(CHOP)的蛋白表达水平以及凋亡相关蛋白B淋巴细胞瘤-2(Bcl-2)、B淋巴细胞瘤-XL(Bcl-XL)和bcl-2相关X蛋白(Bax)表达水平。结果TG以剂量依赖性抑制AE病人T淋巴细胞的增殖,促进AE病人T淋巴细胞凋亡,并且使AE病人T淋巴细胞的BIP(0.50±0.05)%比(0.23±0.04)%和抗凋亡蛋白Bcl-2(1.41±0.04)%比(0.73±0.07)%、Bcl-XL(1.23±0.06)%比(0.51±0.05)%表达下调,CHOP(0.53±0.06)%比(0.81±0.07)%和促凋亡蛋白Bax(0.51±0.01)%比(1.25±0.05)%的表达水平显著增加。结论内质网应激抑制AE病人T淋巴细胞的增殖,促进细胞凋亡,可能通过BIP和CHOP来影响细胞凋亡。
Objective To observe the effect of endoplasmic reticulum stress on the survival and apoptosis of T lymphocytes in AE patients and the role of unfolded protein response(UPR)signaling pathway in the survival and death of AE lymphocytes.Methods Peripheral blood samples of AE patients and healthy donors from the Department of Pediatrics,Shijiazhuang First Hospital from February 2017 to February 2019 were collected.T lymphocytes of AE patients and healthy donors were treated with TG at different concentrations(0,0.5,1,5,10 nM)for 48 h,and then cell viability was tested by cck-8 assay;TG(concentration:5 n M)was used to treat T lymphocytes of AE patients and healthy donors for 48 h,and the apoptosis of untreated and TG treated T lymphocytes was detected by flow cytometry;TG(concentration:5 nM)stimulated T lymphocytes of AE patients and healthy donors for 48 h,and the protein expression levels of UPR marker molecules(Binding immunoglobulin protein,BIP)and(Homologous protein,CHOP),as well as apoptotic related proteins(B-cell lymphoma-2,Bcl-2),(B-cell lymphoma-XL,Bcl-XL)and(Bcl-2 associated X protein,Bax)were detected by Western blot.ResultsTG inhibited the proliferation of T lymphocytes in AE patients in a dose-dependent manner,promoted the apoptosis of T lymphocytes in AE patients,and down-regulated the expressions of BIP(0.50±0.05)%vs.(0.23±0.04)%and anti-apoptotic proteins Bcl-2(1.41±0.04)%vs.(0.73±0.07)%and Bcl-XL(1.23±0.06)%vs.(0.51±0.05)%,and significantly increased the expressions of CHOP(0.53±0.06)%vs.(0.81±0.07)%and pro-apoptotic protein Bax(0.51±0.01)%vs.(1.25±0.05)%.Conclusion Endoplasmic reticulum stress inhibits the proliferation of T lymphocytes in AE patients and promotes apoptosis,which may be influenced by BIP and CHOP.
作者
王丹
刘晓娟
康天
WANG Dan;LIU Xiaojuan;KANG Tian(Department of Pediatrics,Shijiazhuang First Hospital,Shijiazhuang,Hebei 050011,China)
出处
《安徽医药》
CAS
2020年第10期2042-2046,共5页
Anhui Medical and Pharmaceutical Journal
