端粒酶逆转录酶在急性坏死性胰腺炎大鼠中性粒细胞凋亡中的作用  

Role of telomerase reverse transcriptase on cell apoptosis of neutrophils in rats with acute necrotizing pancreatitis

作　　者：任大宾[1] 杨志文[2] Ren Dabin;Yang Zhiwen(Department of Gastroenterology,Songjiang District Central Hospital,Shanghai 201600,China;Department of Pharmacy,Songjiang District Central Hospital,Shanghai 201600,China)

机构地区：[1]上海市松江区中心医院消化内科,上海201600 [2]上海市松江区中心医院药剂科,上海201600

出　　处：《中华胰腺病杂志》2022年第3期196-200,共5页Chinese Journal of Pancreatology

基　　金：上海市自然科学基金(19ZR1448400)。

摘　　要：目的探讨端粒酶逆转录酶(TERE)对急性坏死性胰腺炎(ANP)大鼠中性粒细胞凋亡的影响。方法24只SD大鼠按数字表法随机分为正常对照组、ANP 3 h组、ANP 6 h组、TERT抑制剂BIBR1532干预组(BIBR1532组),每组6只。采用胰胆管逆行注射5%牛磺胆酸钠构建ANP模型,分别在造模后3、6 h收集和分离大鼠外周血中性粒细胞BIBR1532组在造模前腹腔注射2 mg/kg BIBR1532,在造模后3 h处死大鼠,收集外周血中性粒细胞。应用荧光定量PCR法检测中性粒细胞TERT mRNA表达量;蛋白质免疫印迹法检测中性粒细胞TERT蛋白及凋亡相关蛋白Bcl-xL、Bax表达量;流式细胞术检测中性粒细胞的凋亡;ELISA法检测血清炎症因子TNF-α、IL-6水平。取大鼠胰腺组织行病理学检查。结果正常对照组、ANP 3 h组、ANP 6 h组、BIBR1532组大鼠外周血中性粒细胞TERT mRNA表达量分别为1.03±0.26、3.31±1.07、5.21±0.78、1.95±0.49;TERT蛋白表达量分别为0.09±0.03、0.43±0.12、0.58±0.11、0.22±0.07;Bcl-xL蛋白表达量分别为0.19±0.05、0.50±0.07、0.85±0.04、0.40±0.11;Bax蛋白表达量分别为0.29±0.08、0.23±0.03、0.17±0.02、0.43±0.12;细胞凋亡率分别为(10.03±0.74)%、(7.99±0.27)%、(6.65±0.36)%、(22.98±2.86)%。ANP 3、6 h组TERT mRNA和蛋白表达量、Bcl-xL蛋白表达量高于对照组,Bax蛋白表达量、细胞凋亡率低于对照组,而BIBR1532组TERT mRNA和蛋白表达量、Bcl-xL蛋白表达量低于ANP 3 h组,Bax蛋白表达量、细胞凋亡率高于ANP 3 h组,差异均有统计学意义(P值均<0.05)。正常对照组、ANP 3 h组、BIBR1532组血清TNF-α水平分别为(96.67±27.12)、(382.30±46.33)、(206.88±36.42)ng/L,IL-6水平分别为(43.34±14.50)、(134.21±16.13)、(88.06±13.05)ng/L,ANP 3 h组均高于正常对照组,而BIBR1532组均低于ANP 3 h组,差异均有统计学意义(P值均<0.05)。BEBR1532组大鼠胰腺组织病理改变较ANP组明显改善。结论ANP大鼠外周血中性粒细胞TERT表达量显著升高Objective To investigate the effects of telomerase reverse transcriptase(TERT)on neutrophils apoptosis in rats with acute necrotizing pancreatitis(ANP).Methods Twenty-four Sprague Dawley(SD)rats were randomly divided into three groups including control group,ANP(3h、6h)group and TERT inhibitor(BIBR1532)group using random number method with 6 in each group.ANP rats were induced by retrograde injection of 5%sodium taurocholate into the pancreaticcobiliary duct.After 3 and 6 hours,the fresh neutrophils were collected and isolated from peripheral blood of ANP rats.Rats were intraperitoneally injected with 2 mg/kg BIBR1532.After ANP modeling for 3 h,rats were killed and peripheral neutrophils were collected.Subsequently,the expression of TERT mRNA in neutrophils was tested by real-time quantitative PCR;the protein levels of TERT,BCL-xL and Bax were determined by Western blotting;neutrophil apoptosis was detected by flow cytometry;TNF-αand IL-6 were assayed by Elisa;the rat pancreatic tissue was pathologically examined.Results In neutrophils from control group,ANP 3 h group,ANP 6 h group and BIBR1532 group,TERT mRNA was 1.03±0.26,3.31±1.07,5.21±0.78 and 1.95±0.49;TERT protein expression was 0.09±0.03,0.43±0.12,0.58±0.11 and 0.22±0.07;Bcl-xL protein expression was 0.19±0.05,0.50±0.07,0.85±0.04 and 0.40±0.11;Bax protein expression was 0.29±0.08,0.23±0.03,0.17±0.02 and 0.43±0.12;apoptosis rate was 10.03±0.74%,7.99±0.27%,6.65±0.36%and 22.98±2.86%.TERT mRNA and protein expression and Bcl-xL protein expression in ANP 3 h and 6 h group were higher than those in control group,but Bax preotein expression and apoptosis rate were lower than those in control group;TERT mRNA and protein expression and Bcl-xL protein expression in BIBR1532 group were lower than those in ANP 3 h group,but Bax protein expression and apoptosis rate in BIBR1532 group were higher than those in ANP 3 h group;and all the differences were statistically significant(all P value<0.05).The level of TNF-αwas[(96.67±27.12)ng/L,(382.30±46.

关 键 词：胰腺炎 急性坏死性 端粒酶逆转录酶 中性粒细胞 细胞凋亡 

分 类 号：R576[医药卫生—消化系统]