中国普外基础与临床杂志

中国普外基础与临床杂志

雷帕霉素对重症急性胰腺炎胰腺损害的保护性作用

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目的探究雷帕霉素对重症急性胰腺炎(severe acute pancreatitis,SAP)大鼠胰腺损害的保护性作用,并进一步阐述其保护机制。方法将 90 只 SPF 级雄性 SD 大鼠随机分为 3 组:即假手术组(SO 组)、SAP 组及雷帕霉素组(RAPA 组),每组 30 只大鼠;每组大鼠再随机分为 24、36 和 48 h 3 个时点亚组,每个亚组 10 只大鼠。每组大鼠均行开腹手术,采用逆行胰胆管注射法制备模型,SO 组大鼠仅注射 0.9% 生理盐水,SAP 组和 RAPA 组大鼠注射 5% 牛磺胆酸钠溶液,且 RAPA 组在注射 5% 牛磺胆酸钠前 30 min 经腹腔注射雷帕霉素。分别于 24、36 及 48 h 处死相应亚组存活的大鼠,收集大鼠血清和胰腺组织,采用 ELISA 法检测血清 IL-1β、IL-6 及 TNF-α 含量,采用 Western blot 法检测胰腺组织中磷酸化的哺乳动物雷帕霉素靶蛋白(p-mTOR)和磷酸化的核糖体 S6 蛋白激酶 1(p-S6K1)的表达水平;胰腺组织行 HE 染色,光镜下进行病理改变评分。结果① 胰腺组织p-mTOR 和 p-S6K1 的表达水平:24、36 及 48 h 时,胰腺组织中 p-mTOR 和 p-S6K1 的表达水平均是 SO 组<RAPA 组<SAP 组,3 组间两两比较差异均有统计学意义(P<0.05)。② IL-1β:48 h 时,3 组的血清 IL-1β 含量顺序为 SO 组<RAPA 组<SAP 组,3 组间两两比较差异均有统计学意义(P<0.05);IL-6:36 h 和 48 h 时,3 组的血清 IL-6 含量顺序为 SO 组<RAPA 组<SAP 组,3 组间两两比较差异均有统计学意义(P<0.05);TNF-α:48 h 时,3 组的血清TNF-α 含量顺序为 SO 组/RAPA 组<SAP 组(P<0.05),但 SO 组和 RAPA 组比较差异无统计学意义(P>0.05)。③ 胰腺织病理学评分:24、36 及 48 h 时 3 组胰腺组织的病理学评分顺序均为 SO 组<RAPA 组<SAP 组,3 组间两两比较差异均有统计学意义(P<0.05)。④ 胰腺组织中 p-mTOR 和 p-S6K1 的表达水平与胰腺组织的病理学评分均呈正相关(r=0.97,P<0.01;r=0.89,P<0.01)。结论雷帕霉素可以减轻 SAP 的胰腺损害程度,对胰腺组织具有保护性的作用。

ObjectiveTo explore the protective effect of rapamycin on pancreatic damage in severe acute pancreatitis (SAP) and further to explain its protective mechanism.MethodsNinety selected SPF males SD rats were randomly divided into 3 groups: sham-operated group (SO group), SAP group, and rapamycin group (RAPA group), with 30 rats in each group. Then each group of rats were randomly divided into 3 subgroups of 24 h, 36 h, and 48 h, 10 rats in each subgroup. Rats in each group underwent laparotomy, the model was prepared by retrograde injection of solutions into biliopancreatic duct, rats of the SO group were injected with 0.9% normal saline, rats of the SAP group and RAPA group were injected with 5% sodium taurocholate solution, but rats of the RAPA group were injected with rapamycin 30 min before the injection of 5% sodium taurocholate. All the survival rats in corresponding subgroup were killed at 24 h,36 h, and 48 h respectively, then serum and pancreas tissues of rats were collected, serum inflammatory factors content of IL-1β, IL-6, and TNF-α were detected by ELISA method, expression levels of p-mTOR and p-S6K1 in pancreas were detected by Western blot, pancreas tissues were stained by Hematoxylin-Eosin Staining and pathological changes of pancreas were scored under light microscope.Results① At the timepoint of 24 h, 36 h, and 48 h, the order of the expression levels of p-mTOR and p-S6K1 in pancreatic tissues of 3 groups were all as follows: SO group<RAPA group<SAP group, there were significant difference among any 2 groups (P<0.05). ② IL-1β: at the timepoint of 48 h, the order of the content of IL-1β in 3 groups were as follows: SO group<RAPA group<SAP group, there were significant differences among any 2 groups (P<0.05); IL-6: at the timepoint of 36 h and 48 h, the order of the content of IL-6 in3 groups were as follows: SO group<RAPA group<SAP group, there were significant differences among any 2 groups (P<0.05); TNF-α: at the timepoint of 48 h, the order of the content of TNF-α in 3 groups was as follows: SO/RAPA group<SAP group (P<0.05), but there was no significant difference between the SO group and RAPA group (P>0.05). ③ Pancreatic histological score: at the timepoint of 24 h, 36 h, and 48 h, the order of the pancreatic histological score in3 groups was all as follows: SO group<RAPA group <SAP group, there were significant differences among any 2 groups (P<0.05). ④ The expression levels of p-mTOR and p-S6K1 in pancreatic tissue were positively correlated with the pathological scores of pancreatic tissue (r=0.97, P<0.01; r=0.89, P<0.01).ConclusionRapamycin can reduce the degree of pancreatic damage in SAP and has protective effect on pancreatic tissue.

关键词: 重症急性胰腺炎; 雷帕霉素; 胰腺损害; 作用

Key words: severe acute pancreatitis; rapamycin; pancreatic damage; effect

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