Treg及IL-4对重症急性胰腺炎时枯否细胞M2极化状态的调节作用

Effect of CD4^+CD25^+Fox P_3^+ regulatroy T cells and IL-4 on kupffer cell M2 polarization state of severe acute pancretitis

目的探讨雨蛙肽联合脂多糖诱导小鼠重症急性胰腺炎(severe acute pancreatitis,SAP)的方法和Treg(CD4+CD25+FoxP3+T调节淋巴细胞)及IL-4(白介素-4)对SAP时枯否细胞(Kupffer cells)M2极化状态的调节作用的比较。方法 (1)正常组予腹腔注射生理盐水(NS);SAP组予腹腔注射雨蛙肽联合脂多糖。SAP组细分为3组:造模后9 h、12 h和24 h组,比较胰腺病理情况。(2)比较正常组与模型组(SAP 8 h+NS组)肝脏炎症因子的基因表达水平。(3)模型组分3组:SAP生理盐水对照组(SAP16 h+NS组)、SAP IL-4治疗组(SAP 16 h+IL-4组)、SAP Treg治疗组(SAP 16 h+Treg组)。RT-PCR检测肝脏炎症因子IL-1β、TNF-α和IL-10 mRNA表达水平;免疫荧光技术检测肝脏CD163、CCR7的表达。结果 (1)HE病理结果造模后24 h可见胰腺片状坏死。(2)模型组IL-1βmRNA的表达显著高于正常组(P<0.05)。(3)SAP Treg治疗组IL-1β、TNF-αmRNA的表达显著低于SAP生理盐水对照组(P<0.05);SAP Treg治疗组IL-1βmRNA的表达显著低于SAP IL-4治疗组(P<0.05)。结论雨蛙肽联合脂多糖成功诱导小鼠SAP。IL-4和Treg对SAP具有一定的治疗作用。

Objective To investigate the effective method of inducing severe acute pancreatitis （SAP） with cerulein plus lipopolysaccharide （LPS） and the regulation of the M2 anti-inflammatory kupffer cells by IL-4 and CD4 ^＋ CD25^ ＋FoxP3^ ＋ regulatory T cells on SAP in mice.Methods （1） Normal group was induced by intraperitoneal injection of saline ; SAP group was induced by intraperitoneal injection of cerulein plus LPS.SAP group was divided into three groups：9 hours,12 hours and 24 hours after making mold groups.Histopathological alterations of pancreatic tissues were evaluated between these three groups and the normal group.（2） Expression of inflammatory cytokines mRNA in liver tissues were assessed by RT-PCR between normal group and SAP 8 h ＋ NS group.（3） SAP models were divided into three groups：SAP 16 h ± NS group,SAP 16 h ＋ IL-4 group and SAP 16 h ± Treg group.Expression of IL-1 β,TNF-α and IL-10 mRNA in liver were assessed by RT-PCR; expression of CD163 and CCR7 in liver were assessed by confocal immunofluorescence microscopy.Results （1） The results of HE staining：acinar cell necrosis was found in 24 hours after making mold.（2） The expression of IL-1 β of model group was significantly higher than that of normal group （P 〈0.05）.（3） The expression of IL-1 β、TNF-α mRNA of SAP 16 h ＋ Treg group were significantly lower than that of SAP 16 h ＋ NS group （P 〈 0.05）.Conclusion It is a effective method by cerulein plus LPS to induce SAP in mice.IL-4and Treg have therapeutic effects on SAP.