STAT3/IL-21通路介导长链非编码RNA H19参与大鼠重症急性胰腺炎相关性肠损伤

Effect of LncRNA H19 up-regulation mediated by STAT3/IL-21 pathway on intestinal injury in rats with severe acute pancreatitis

目的探讨长链非编码RNA(LncRNA)H19及STAT3/IL-21通路在大鼠重症急性胰腺炎(SAP)发生发展中的作用。方法45只SD大鼠分为对照组(n=15)、SAP组(n=15)和转录激活因子3(STAT3)抑制组(n=15)。SAP组大鼠经胰胆管逆行微量泵入5%牛磺胆酸钠(0.1 mL/100 g)制备SAP模型。对照组大鼠泵入生理盐水。STAT3抑制组大鼠在诱导SAP后经腹腔注射STAT3抑制剂(STAT3-IN-1),剂量为10 mg/kg。其余两组注射等体积0.5%DMSO。造模后24 h处死大鼠,留取血清及肠道组织标本。酶联免疫吸附法(ELASA)检测大鼠血清淀粉酶及IL-21水平,RT-PCR法测定各组大鼠血清中H19和STAT3基因mRNA表达水平,以及测定各组大鼠肠道组织中带状闭合蛋白-1(ZO-1)、咬合蛋白(Occludin)、闭合蛋白(Claudin)-1和Claudin-2基因mRNA表达差异。结果与对照组比较,两个实验组大鼠血清中淀粉酶表达明显升高(F=51.120,P<0.01),证明造模成功;SAP组大鼠血清IL-21较对照组明显升高,STAT3被抑制后,IL-21呈下降趋势(F=40.220,P<0.01)。SAP组大鼠血清中H19、STAT3基因mRNA表达均明显升高,而与SAP组比较,STAT3抑制组两基因表达水平均呈降低趋势(H19:F=15.165,STAT3:F=31.279,P<0.01)。此外,与对照组比较,SAP组大鼠肠道组织中ZO-1、Occludin、Claudin-1基因mRNA表达明显降低,而Claudin-2基因表达明显升高;与SAP组比较,STAT3抑制剂组ZO-1、Occludin、Claudin-1基因mRNA表达均呈下降趋势,Claudin-2基因表达明显升高(ZO-1:F=31.478,Occludin:F=24.614,Claudin-1:F=13.839,Claudin-2:F=11.496,P<0.01)。结论大鼠SAP相关性肠屏障功能障碍发生发展可能与STAT3/IL-21通路介导的H19表达上调有关。

Objective To explore the possible role of long non-coding RNA(LncRNA)H19 and signal transducer and activator of transcription 3(STAT3)/interleukin-21(IL-21)pathway in the occurrence and development of the severe acute pancreatitis(SAP).Methods A total of 45 SD rats were randomly divided into control group(n=15),SAP group(n=15)and STAT3 inhibition group(n=15).SAP group received retrograde pumping of 5%sodium taurocholate(0.1 mL/100 g)into the pancreaticobiliary duct to induce SAP;the rats in the control group were injected with normal saline.STAT3 inhibition group were intraperitoneally injected with a STAT3 inhibitor(STAT3-IN-1)at the dose of 10 mg/kg,and the other two groups were injected with equal volume 0.5%DMSO.At 24 h after modeling,all the rats were executed,and the serum and the intestinal tissues were collected.The levels of IL-21 and amylase in serum were determined by enzyme-linked immunosorbent assay(ELISA).The expression levels of STAT3 mRNA and H19 mRNA in serum were determined by RT-PCR.The mRNA expression levels of ZO-1,Occludin,Claudin-1 and Claudin-2 in the intestinal tissues were detected.Results Compared with the control group,the expression levels of serum amylase in the two experimental groups significantly increased(F=51.120,P<0.01).The expression levels of IL-21 in the SAP group significantly increased,IL-21 significantly decreased after STAT3 was inhibited(F=40.220,P<0.01).The mRNA expression levels of H19 and STAT3 genes increased significantly in the SAP group,but the expression levels of two genes showed a downward trend in STAT3 inhibition group compared with SAP group(H19:F=15.165,STAT3:F=31.279,P<0.01).In addition,compared with the control group,the mRNA expression levels of ZO-1,Occludin,and Claudin-1 genes in the intestinal tissues of the SAP group were significantly reduced,while the expression levels of Claudin-2 genes were significantly increased,the mRNA expression levels of ZO-1,Occludin and Claudin-1 genes showed a downward trend in STAT3 inhibition group,and the expression levels of Claudin-2 genes increased significantly(ZO-1:F=31.478,Occludin:F=24.614,Claudin-1:F=13.839,Claudin-2:F=11.496,P<0.01).Conclusions The development of intestinal barrier dysfunction in the rats with SAP may be related to the up-regulation of H19 expression mediated by STAT3/IL-21 pathway.