纤维粘连蛋白靶向JAK/STAT3信号通路调控粘连性肠梗阻的机制

Mechanism of fibronectin targeting JAK/STAT3 signaling pathway to regulate adhesive ileus

目的:探究纤维粘连蛋白(FN)靶向JAK/STAT3信号通路调控粘连性肠梗阻的作用机制。方法:选取2019年1月—2020年12月就诊行肠切除手术的粘连性肠梗阻患者65例(观察组),同期行腹股沟疝手术患者65例(对照组),取患者肠组织及术前血液样本,采用qRT-PCR、Western blot及免疫组织化学实验检测肠组织样本中FN、STAT3、p-STAT3基因及蛋白表达,采用酶联免疫吸附实验(ELISA)检测血清样本中FN表达。体外培养人肠上皮细胞系HIEC-6,采用慢病毒转染抑制细胞FN表达,采用CCK8、Transwell迁移实验观察FN对细胞增殖及分化的影响,同时采用qRT-PCR、Western Blot实验检测FN对细胞STAT3、p-STAT3表达的影响。结果:与对照组比较,观察组肠组织中FN mRNA及蛋白表达明显较高(P<0.05),STAT3、p-STAT3 mRNA及蛋白表达则明显较低(P<0.05),且观察组血清中分泌型FN含量也明显较高(P<0.05)。体外实验显示,与NC siRNA组HIEC-6细胞比较,LV-FN siRNA组HIEC-6细胞在48、72、96、120 h的增殖率均明显较低(P<0.05),细胞迁移能力也明显较低(P<0.05),其FN mRNA及蛋白表达明显较低(P<0.05),STAT3、p-STAT3 mRNA及蛋白则明显较高(P<0.05)。结论:纤维粘连蛋白在粘连性肠梗阻患者肠组织及血清中呈明显高表达,可通过JAK/STAT3信号通路的异常激活影响细胞的增殖和迁移,引起病变肠段发病。

Objective:To explore and analyze the mechanism of Fibronectin(FN)targeting JAK/STAT3 signaling pathway to regulate adhesive ileus.Methods:From January 2019 to December 2020,65 patients with adhesive ileus who underwent intestinal resection were selected as the observation group,and 65 patients who underwent inguinal hernia surgery during the same period were selected as the control group.Intestinal tissue and preoperative blood samples were taken from the patients.qRT-PCR,Western Blot and immunohistochemistry were used to detect FN,STAT3 and P-STAT3 gene and protein expression in intestinal tissue samples,and Enzymelinked immunosorbent assay(ELISA)was used to detect FN expression in serum samples.Cell line HIEC-6 was cultured in vitro.FN expression was inhibited by lentivirus transfection,and the effects of FN on cell proliferation and differentiation were observed by CCK8 and Transwell migration experiments.At the same time,qRT-PCR and Western Blot were used to detect the effects of FN on the expression of STAT3 and P-STAT3.Results:Compared with the control group,the expression of FN mRNA and protein in intestinal tissue of observation group was significantly higher(P<0.05),while the expression of STAT3 and P-STAT3 mRNA and protein was significantly lower(P<0.05),and the content of secreted FN in serum of observation group was also significantly higher(P<0.05).In vitro experiments showed that compared with NC siRNA group HIEC-6 cells,the proliferation rate of LV-FN siRNA group HIEC-6 cells was significantly lower at 48 h,72 h,96 h and 120 h(P<0.05),and the cell migration ability was also significantly lower(P<0.05).The mRNA and protein expressions of FN were significantly lower(P<0.05),while the mRNA and protein expressions of STAT3 and P-STAT3 were significantly higher(P<0.05).Conclusion:Fibroinin is significantly overexpressed in the intestinal tissues and serum of patients with adhesive ileus,and can affect cell proliferation and migration through abnormal activation of JAK/STAT3 signaling pathway,leading to the pathogenesis of the diseased intestinal segment.