拓扑异构酶I抑制剂喜树碱对小鼠急性胰腺炎保护机制的研究  

Molecular mechanism of topoisomerase I inhibitor in protecting against experimental acute pancreatitis in mice

作　　者：包静飘 李彬[1] 吴江红 吴增楷 沈杰 宋鹏丽 彭琪 胡国勇 Bao Jingpiao;Li Bin;Wu Jianghong;Wu Zengkai;Shen Jie;Song Pengli;Peng Qi;Hu Guoyong(Shanghai Key Laboratory of Pancreatic Diseases,Department of Gastroenterology,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200080,China)

机构地区：[1]上海市胰腺疾病重点实验室,上海交通大学医学院附属第一人民医院消化科,上海200080

出　　处：《中华胰腺病杂志》2022年第2期107-112,共6页Chinese Journal of Pancreatology

基　　金：国家自然科学基金(81970556、82170652)。

摘　　要：目的探讨拓扑异构酶I抑制剂喜树碱对AP小鼠胰腺腺泡细胞和肺组织的保护机制。方法18只Balb/C雄性小鼠按照数字表法随机分为对照组、AP组和喜树碱干预组(喜树碱组)。AP组采用腹腔注射雨蛙肽和脂多糖的方法制备AP模型,喜树碱组于制模前腹腔注射50 mg/kg喜树碱,对照组腹腔注射等体积生理盐水。常规行胰腺和肺组织病理学检查。分别采用ELISA法检测血清淀粉酶和脂肪酶水平;荧光定量聚合酶链式反应(RT-PCR)检测胰腺组织炎症因子IL-1、IL-6 mRNA表达量;免疫组织化学法检测胰腺和肺组织CD_(45)^(+)炎症细胞浸润以及胰腺组织程序性坏死标志蛋白混合谱系激酶结构域样蛋白(MLKL)磷酸化水平;TUNEL法检测胰腺组织细胞凋亡指数。结果喜树碱组的胰腺组织病理评分、肺组织病理评分、血清淀粉酶和脂肪酶含量分别为(2.30±0.31)分、(2.29±0.34)分、(1742.33±183.51)U/L和(46.90±2.17)U/L,均显著低于AP组的(5.06±0.88)分、(3.40±0.09)分、(2385.33±383.10)U/L和(69.13±9.76)U/L;胰腺组织IL-1、IL-6 mRNA表达量分别为95.79±48.11、255.50±213.32,均显著低于AP组的212.35±80.61、1006.80±509.06;胰腺和肺组织CD45+细胞数分别为(14.25±5.32)、(29.20±4.44)个/高倍视野,均显著低于AP组的(59.83±13.67)、(58.25±5.91)个/高倍视野。与AP组比较,喜树碱组胰腺组织细胞凋亡指数显著升高[(3.64±1.16)%比(1.92±0.29)%],磷酸化MLKL蛋白免疫组织化学评分显著降低[(1.75±0.20)分比(4.53±1.28)分],差异均有统计学意义(P值均<0.05)。结论拓扑异构酶I抑制剂喜树碱可能通过诱导胰腺腺泡细胞凋亡并抑制腺泡细胞坏死重塑AP小鼠腺泡细胞死亡方式,进而减轻胰腺局部损伤及急性胰腺炎相关肺损伤。Objective To investigate the protective mechanism of topoisomerase I inhibitor on pancreatic acinar cells and lung during acute pancreatitis(AP)in mice.Methods Eighteen Balb/C male mice were randomly divided into three groups using random number method:control group,AP group and CPT+AP group.AP model was established by intraperitoneal injection of caerulein and lipopolysaccharide.CPT+AP group received intraperitoneal injection of camptothecin(CPT,50 mg/kg)before AP induction.Mice in control group were intraperitoneal injected with an equal volume of normal saline.The pathological examinations of pancreas and lung tissue were analyzed.The serum levels of amylase and lipase were detected by enzyme linked immunosorbent assay(ELISA)and the mRNA expression of IL-1 and IL-6 were analyzed by reverse transcription-polymerase chain reaction(RT-PCR);the infiltration of CD_(45)^(+)cells in pancreas and lung tissue as well as the expression of phosphorylated mixed lineage kinase domain-like protein(MLKL)in pancreas were detected by immunohistochemistry;the apoptosis index of pancreatic cells was analyzed by TUNEL assay.Results The pathological scores of pancreas and lung tissue,serum levels of amylase and lipase in CPT+AP group were[(2.30±0.31),(2.29±0.34),(1742.33±183.51)U/L and(46.90±2.17)U/L],which were significantly lower than those in AP group[(5.06±0.88),(3.40±0.09),(2385.33±383.10)U/L and(69.13±9.76)U/L];the mRNA expression of IL-1 and IL-6 in pancreatic tissue were 95.79±48.11,255.50±213.32,which were also remarkably lower than those in AP group(212.35±80.61,1006.80±509.06);the infiltration of CD45+inflammatory cells in pancreas and lung were(14.25±5.32,29.20±4.44)/high power field,which were notably lower than those in AP group(59.83±13.67,58.25±5.91)/high power field;the apoptosis index of pancreatic cells was significantly higher than that in AP group[(3.64±1.16)%vs(1.92±0.29)%];the histochemistry score of phosphorylated MLKL protein in pancreatic tissue was significantly lower than that in AP gr

关 键 词：拓扑异构酶I抑制剂 胰腺炎 细胞凋亡 细胞死亡 小鼠 

分 类 号：R576[医药卫生—消化系统]