青蒿琥酯对重型急性胰腺炎大鼠胰腺细胞凋亡的保护作用  

A Study of Protective Effect of Artesunate on Pancreatic Cell Apoptosis in Rats with Severe Acute Pancreatitis

作　　者：李思伟 王素彬[1] 周江帆 张明谦[1] 顾松涛 杨明东 杨睿[1] 易敬帅 张志华 LI Si-wei;WANG Su-bin;ZHOU Jiang-fan;ZHANG Ming-qian;GU Song-tao;YANG Ming-dong;YANG Rui;YI Jing-shuai;ZHANG Zhi-hua(Dept.of Emergency,Yan＇an Hospital Affiliated of Kunming Medical University,Kunming Yunnan 650051,China)

机构地区：[1]昆明医科大学附属延安医院急诊医学部,云南昆明650051

出　　处：《昆明医科大学学报》2018年第10期7-12,共6页Journal of Kunming Medical University

基　　金：国家自然科学基金资助项目(81560140);昆明市延安医院院内基金资助项目(yyky017-003)

摘　　要：目的探讨青蒿琥酯对大鼠重型胰腺炎时胰腺细胞凋亡的保护作用。方法逆行胆管微量泵入5%牛磺酸钠制备大鼠急性重型胰腺炎模型,鼠尾静脉采血应用酶联免疫吸附(ELASE)方法检测血清淀粉酶,肿瘤坏死因子-α(TNF-α),白介素-6(IL-6)及髓过氧化物酶(MPO),丙二醛(MDA),超氧化物歧化酶(SOD)的变化情况;用Western-bolt蛋白印迹分析方法检测大鼠胰腺组织内凋亡相关因子半胱天冬酶-3(Caspase-3)和半胱天冬酶-9(caspase-9)的表达水平;对大鼠胰腺病理切片伊红(HE)染色观察大鼠胰腺组织坏死,出血,炎症,水肿等情况,并进行病理学组织学分析,原位末端标记法(TUNEL)检测大鼠胰腺细胞凋亡情况及凋亡指数,根据各组大鼠存活率绘制生存曲线并进行比较。结果大鼠血清淀粉酶在重型胰腺炎组大鼠(Severe acute pancreatitis,SAP)与假手术组比较明显增高(P<0.05),青蒿琥酯干预处理后血清淀粉酶水平较SAP组大鼠有明显下降趋势(P<0.05);对各组实验大鼠血清中炎症因子TNF-α, IL-6, MPO, MDA检测发现经青蒿琥酯干预处理后较SAP组明显下降(P<0.05)而SOD水平升高(P<0.05);通过对胰腺组织中凋亡蛋白Caspase-3和Caspase-9蛋白表达水平进行检测发现青蒿琥酯干预处理后较SAP组其表达水平下降(P<0.05);大鼠胰腺病理学切片HE染色后观察发现青蒿琥酯干预后与SAP组比较胰腺坏死水肿等情况有所缓解,病理学评分下降(P<0.05);应用TUNEL法检测胰腺细胞凋亡情况显示青蒿琥酯干预后胰腺细胞凋亡减少(P<0.05),生存曲线分析发现青蒿琥酯干预后大鼠存活率增加(P<0.05)。结论青蒿琥酯可延缓急性重型胰腺炎大鼠胰腺细胞凋亡,增加大鼠生存率,有重要临床应用价值。Objective To investigate the protective effect of artesunate on the apoptosis of pancreatic cells inrats with severe pancreatitis. Methods The rat model of acute severe pancreatitis was prepared by retrogrademicroinfusion of sodium taurine into 5% sodium taurine. The blood serum amylase,tumor necrosis factor - alpha（TNF- alpha ）, interleukin -6 （IL-6 ） and myeloperoxidase（MPO ）, the changes of malondialdehyde（MDA ） and superoxide dismutase （SOD ） were detected by enzyme linked immunosorbent assay （ELASE） inthe tail vein of rat. The expression of caspase -3 （caspase-3） and caspase -9 （caspase-9） in the pancreatictissue of rats was detected by Western-bolt blot analysis. The necrosis,bleeding,and inflammation of the pancreastissue were observed in rat pancreas pathological section of eosin （HE） staining. The rat pancreatic cell apoptosis and apoptosis index were detected by in situ end labeling method （TUNEL ）,and the survival curves were plottedaccording to the survival rate of rats in each group and compared.Results The serum amylase in the rats of thesevere pancreatitis group （severe acute pancreatitis SAP ） was significantly higher than that in the sham group（ P〈0.05） . After the treatment of artesunate,the serum amylase level was significantly lower than that of the SAPgroup （P 〈0.05）,and the serum levels of inflammatory factors TNF-a,IL-6,MPO,MDA were found to besignificantly decreased after the treatment of artesunate （ P〈0.05 ） and SOD level （ P〈0.05） . The expressionlevel of artesunate was detected and the expression level of artesunate was decreased （P 〈0.05） . After HE stainingin the pathological section of the pancreas of rats,the results showed that the prognosis of artesunate was less thanthat of the SAP group. The pathological score of the pancreas was decreased （ P〈0.05）,and the apoptosis ofpancreatic cells by TUNEL method showed that the pancreas was fine after artesunate intervention. The decrease ofapoptosis （ P〈0

关 键 词：青蒿琥酯 急性重症胰腺炎 细胞凋亡 

分 类 号：R576[医药卫生—消化系统]