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目的探讨促肾上腺皮质激素释放因子(CRF)对肠道杯状细胞分泌产物的影响。方法首先用不同浓度的CRF作用于LS174T细胞以确定最佳作用浓度;用10~(-10)mol/L的CRF作用于LS174T细胞,利用Real-time q PCR、Western blotting、免疫荧光染色和ELISA技术对肠道杯状细胞分泌产物的表达量进行检测。结果 Realtime q PCR结果显示,与PBS对照组相比,CRF作用6 h后,黏蛋白2(M UC2),岩藻糖基转移酶-2(FUT2)和三叶因子-3(TFF3)的mRNA表达量均下降(P<0.05)。细胞免疫荧光和ELISA的结果显示,CRF作用6 h后,MUC2的蛋白表达量降低(P<0.05)。Western blotting结果显示,CRF作用24 h后,FUT2和TFF3的蛋白表达水平均下降(P<0.05)。结论 CRF可以通过改变LS174T细胞分泌产物的表达,从而破坏肠黏膜屏障功能。
Objective To investigate the effect of corticotropin releasing factor (CRF) on the secretion of intestinal goblet cells. Methods LS174T cells were treated with different concentrations of CRF to determine the best concentration. The cells were treated with 10 ~ (-10) mol / L CRF in LS174T cells. Real-time qPCR, Western blotting, immunofluorescence and ELISA Technology to detect the quantity of intestinal goblet cell secretory products. Results Realtime qPCR results showed that mRNA expression of MUC2, FUT2 and TFF3 mRNA increased after 6-hour CRF treatment compared with PBS control group The expression levels decreased (P <0.05). Cell immunofluorescence and ELISA showed that the expression of MUC2 decreased after 6 hours of CRF treatment (P <0.05). Western blotting showed that the protein expression of FUT2 and TFF3 decreased after 24 h of CRF treatment (P <0.05). Conclusion CRF can affect the intestinal mucosal barrier function by changing the expression of secreted products of LS174T cells.