Resistin促进急性胰腺炎病情加重的作用机制

Mechanism of Resistin in Promoting Exacerbation of Acute Pancreatitis

目的分析抵抗素(Resistin)对雨蛙素(caerulein,CN)诱导的小鼠重症急性胰腺炎(SAP)模型的影响,并探讨其恶化机制。方法将BALB/C小鼠随机分为5组:对照组、CN组、Resistin+CN组、sh-Resistin+CN组和Resistin+CN+3-MA组,每组10只。对Resistin+CN组、sh-Resistin+CN组和Resistin+CN+3-MA组小鼠施用携带Resistin、sh-Resistin的AVV病毒载体;除对照组外,其他4组通过以1 h间隔腹膜内注射10次CN(100μg·kg^(-1))诱导CN胰腺炎模型;Resistin+CN+3-MA组在最后1次CN注射后2 h通过腹膜内注射3-MA(20 mg·kg^(-1))。采用干化学法测量血清淀粉酶、酶比色法检测脂肪酶活性。胰腺切片苏木精和伊红(HE)染色后进行胰腺组织病理学评分,采用TUNEL法测定胰腺组织细胞凋亡、透射电子显微镜观察胰腺自噬、Western Blot检测胰腺组织中Resistin表达。结果与CN组相比,Resistin+CN组的血清淀粉酶、脂肪酶活性,胰腺组织病理学评分和TUNEL阳性细胞百分比显著增加(P<0.05);sh-Resistin+CN组小鼠的血清淀粉酶、脂肪酶活性,胰腺组织病理学评分和TUNEL阳性细胞百分比显著降低(P<0.05)。与CN组相比,Resistin+CN组中每个细胞质区域的自噬泡百分比显著增加,sh-Resistin+CN组的自噬泡百分比显著降低。Resistin+CN组胰腺组织中P62、LC3Ⅱ水平较CN组显著增加,而sh-Resistin+CN组胰腺组织中P62、LC3Ⅱ水平较CN组显著降低。与Resistin+CN组相比,Resistin+CN+3-MA组的胰腺组织学评分以及胰腺组织中P62、LC3Ⅱ水平和自噬泡百分比均显著降低(P<0.05)。结论Resistin通过诱导自噬受损在SAP发展中发挥重要作用。

Objective To investigate the role of resistin in a mouse model of severe acute pancreatitis(SAP)induced by cerulein(CN),and to characterize its regulatory mechanism.Methods BALB/C mice were randomly divided into 5 groups:control group,CN group,resistin+CN group,sh-resistin+CN group and resistin+CN+3-MA group,with 10 mice in each group.Adenoviral vectors carrying resistin or sh-resistin were administered to the mice in resistin+CN group,sh-resistin+CN group and resistin+CN+3-MA group.Except the control group,the other 4 groups were intraperitoneally injected with CN(100μg·kg^(-1))10 times at 1-h interval to induce pancreatitis.In resistin+CN+3-MA group,3-MA(20 mg·kg^(-1))was administered by intraperitoneal injection 2 h after the last CN injection.Serum amylase was measured by dry chemistry,and lipase activity was detected by enzymatic colorimetry.Pancreatic sections were stained with hematoxylin and eosin(HE)for histopathological scoring.Pancreatic apoptosis was observed by TUNEL assay,and pancreatic autophagy by transmission electron microscopy.Resistin expression in pancreatic tissue was detected by Western blot.Results Compared with CN group,serum amylase,lipase activity,pancreatic histopathological score,percentage of TUNEL-positive cells,percentage of autophagic vesicles,and expression of P62 and LC3Ⅱincreased in resistin+CN group,but decreased in sh-resistin+CN group(P<0.05).Compared with resistin+CN group,pancreatic histological score,levels of P62 and LC3Ⅱand percentage of autophagic vesicles decreased in resistin+CN+3-MA group(P<0.05).Conclusion Resistin plays an important role in SAP development by inducing autophagy.