PD-1抗体对PM2.5诱导的大鼠心血管及血管内皮细胞损伤的影响  

Effect of PD-1 antibody on PM2.5-induced damage of cardiovascular and vascular endothelial cells in rats

作　　者：刘振[1] 韩明磊[1] 崔佳佳 侯永兰[1] 徐光翠 LIU Zhen;HAN Ming-Lei;CUI Jia-Jia(Department of Cardiology,Xinxiang Central Hospital,Xinxiang 453000,Henan,China)

机构地区：[1]新乡市中心医院心血管内科,河南新乡453000 [2]新乡医学院基础医学院

出　　处：《中国老年学杂志》2023年第11期2709-2715,共7页Chinese Journal of Gerontology

基　　金：国家青年科学基金项目(81703183);河南省医学科技攻关联合共建项目(LHGJ20200937)。

摘　　要：目的探讨程序性死亡蛋白(PD)-1抗体对PM2.5染毒大鼠心血管损伤的作用及对PM2.5引起人血管内皮细胞损伤的影响。方法40只雄性SD大鼠随机分成对照组、PM2.5组、低剂量(PM2.5+低剂量PD-1抗体)组、高剂量(PM2.5+高剂量PD-1抗体)组,每组10只。气管滴注PM2.5悬液进行染毒,每隔2 d滴注1次,持续3 w,并以低、高剂量PD-1抗体(10 mg/kg、20 mg/kg)注射腹腔。实验前后测定大鼠体质量;末次染毒24 h后,腹主动脉取血,检测血清中氧化应激和炎症反应相关因子的水平;苏木素-伊红(HE)染色检测肺部和心脏组织形态学变化;TUNEL染色检测心肌细胞凋亡情况。人血管内皮细胞EA.hy926随机分为对照组、PM2.5组、低剂量组、高剂量组,PM2.5组用含100μg/ml PM2.5培养液培养,低、高剂量组用含1 mg/ml PM2.5培养液和PD-1抗体(0.5、1.0 mg/ml)培养,24 h后收集细胞,噻唑蓝(MTT)法和流式细胞术检测细胞活性和凋亡;Western印迹法检测活化型半胱氨酸天冬氨酸蛋白酶(Cleaved caspase)-3、Cleaved caspase-9蛋白表达。结果与对照组比较,实验后PM2.5组大鼠体质量减轻,血清中超氧化物歧化酶(SOD)活性、谷胱甘肽(GSH)含量下降,丙二醛(MDA)、白细胞介素(IL)-6、肿瘤坏死因子(TNF)-α、内皮素(ET)-1和C反应蛋白(CRP)含量均升高,肺组织和心肌组织出现明显的病理改变,心肌细胞凋亡增加,差异均具有统计学意义(P<0.01)。与PM2.5组比较,经高、低剂量PD-1抗体处理的大鼠,体质量有所增加,血清中SOD活性及GSH含量升高,MDA、IL-6、TNF-α及CRP含量均下降,高剂量PD-1抗体处理下ET-1也下降,肺组织和心肌组织病理学形态得到改善,心肌细胞凋亡减少,差异均具有统计学意义(P<0.01)。与对照组比较,PM2.5组EA.hy926细胞存活率降低,细胞凋亡率增加,Cleaved caspase-3和Cleaved caspase-9表达增加,差异均具有统计学意义(P<0.01);与PM2.5组比较,低剂量组、高剂量组的细胞存活率均升高,�Objective To investigate the effect of programmed death protein(PD)-1 antibody on the cardiovascular injury in rats exposed to PM2.5,and the effect of PM2.5 on the injury of human vascular endothelial cells.Methods Forty male SD rats were randomly divided into control group,PM2.5 group,low-dose(PM2.5+low-dose PD-1 antibody)group,high-dose(PM2.5+high-dose PD-1 antibody)group,with 10 rats in each group.The PM2.5 suspension was instilled into the trachea for poisoning,once every 2 d for 3 w,low and high doses of PD-1 antibody(10 mg/kg,20 mg/kg)were injected into the intraperitoneal cavity.The body mass of rats was measured before and after the experiment;24 h after the last exposure,blood was taken from the abdominal aorta to detect the levels of oxidative stress and inflammation-related factors in the serum;hematoxylin-eosin(HE)staining was used to detect the morphological changes of lung and heart tissue;TUNEL staining was used to detect the apoptosis of cardiomyocytes.Human vascular endothelial cells EA.hy926 were randomly divided into control group,PM2.5 group,low-dose group and high-dose group,PM2.5 group contained 100μg/ml PM2.5 culture medium,low and high-dose groups were cultured with 1 mg/ml PM2.5 medium and PD-1 antibody(0.5,1.0 mg/ml).The cells after 24 h were collected,thiazole blue(MTT)method and flow cytometry to detect cell viability and apoptosis;Western blot method was used to detect the expression of activated cysteine protease(Cleaved caspase)-3 and Cleaved caspase-9 protein.Results Compared with the control group,the rats lost body mass,activity of serum superoxide dismutase(SOD)and content of glutathione(GSH)were decreased,contents of malondialdehyde(MDA),interleukin(IL)-6,tumor necrosis factor(TNF)-α,endothelin(ET)-1 and C-reactive protein(CRP)were increased,lung and myocardial tissues showed obvious pathological changes,and myocardial cell apoptosis was increased in PM2.5 group after the experiment,the differences were statistically significant(P<0.01).Compared with PM2.5 group,high and low

关 键 词：心血管损伤 PM2.5 程序性死亡蛋白(PD)-1抗体 血管内皮细胞 

分 类 号：R54[医药卫生—心血管疾病] X513[医药卫生—内科学]