安肠愈疡汤对溃疡性结肠炎大鼠肠黏膜屏障的修复作用及机制研究

Research of repair effect and mechanism of Anchang Yuyang decoction on intestinal mucosal barrier in ulcerative colitis rats

关键词:  安肠愈疡汤  溃疡性结肠炎  咬合蛋白  紧密连接蛋白1  白介素-13/酪氨酸激酶/信号转导与转录激活因子6信号通路

Key words:  Anchang Yuyang decoction  ulcerative colitis  Occludin  Claudin-1  IL-13/JAK1/STAT6 signaling pathway

摘要:

[目的] 观察中药安肠愈疡汤对溃疡性结肠炎（UC）大鼠咬合蛋白（Occludin）、紧密连接蛋白1（Claudin-1）基因、蛋白及白细胞介素-13（IL-13）/酪氨酸激酶1（JAK1）/信号转导与转录激活因子6（STAT6）信号通路的调控作用，探讨其修复肠黏膜屏障的机制。[方法] 将36只SD大鼠随机分为6组，分别为空白组、模型组、美沙拉嗪对照组、安肠愈疡汤低、中、高剂量组，经4.5%葡聚糖硫酸钠（DSS）溶液造模及药液灌胃后处死大鼠，留取标本，苏木精-伊红（HE）染色检测各组大鼠结肠黏膜组织病理情况，逆转录聚合酶链反应（RT-PCR）法、蛋白免疫印迹（Western Blot）法检测结肠组织Occludin、Claudin-1基因、蛋白的表达，RT-PCR法检测IL-13、JAK1、STAT6基因的表达。[结果] 各用药组均明显降低组织病理学评分，差异均具有统计学意义（P<0.01），镜下表现为结肠组织结构好转，炎细胞浸润减轻，均可上调Occludin、Claudin-1基因及蛋白表达（P<0.01），差异均具有统计学意义；除低剂量组外，余用药组不同程度激活IL-13/JAK1/STAT6通路的基因表达（P<0.05或P<0.01）；以中药高剂量组和美沙拉嗪组的作用最为显著，差异均具有统计学意义（P<0.05或P<0.01）。相关性分析发现，Occludin、Claudin-1表达水平与JAK1、STAT6表达水平呈负相关（r=-0.664，r=-0.654，r=-0.629，r=-0.637，P<0.05），与IL-13表达水平呈正相关（r=0.888，r=0.983，P<0.05），差异均具有统计学意义。[结论] 中药安肠愈疡汤能够明显促进UC大鼠肠黏膜修复，其作用机制可能与激活IL-13/JAK1/STAT6信号通路，上调Occludin和Claudin-1的表达水平有关。

Abstract:

[Objective] To observe the effect of Anchang Yuyang decoction on Occludin, Claudin-1 gene, protein and IL-13/JAK1/STAT6 signal pathway in ulcerative colitis rats, and to explore the mechanism of repairing intestinal mucosal barrier.[Methods] The 36 SD rats were randomly divided into 6 groups:blank group, model group, mesalazine group and Anchang Yuyang decoction low, medium and high dose group. After 4.5% DSS solution was used to establish the model and the rats were killed by gavage. The pathological conditions of colonic mucosa were examined by HE. The expressions of Occludin, Claudin-1 gene and protein were detected by RT-PCR and Western Blot, and the gene expressions of IL-13, JAK1, STAT6 were detected by RT-PCR.[Results] The score of histopathology was significantly decreased in all treatment groups (P<0.01). The colonic structure is improved and inflammatory cell infiltration was reduced, also Occludin, Claudin-1 gene and protein expression was raised in the treatment group (P<0.01). The gene expression of IL-13/JAK1/STAT6 pathway was activated to different degrees in other groups except the low-dose group(P<0.05 or P<0.01). The high-dose group and the mesalazine group had the most significant effects (P<0.05 or P<0.01). The correlation analysis showed that there was a negative correlation between the expression levels of Occludin, Claudin-1 and JAK1, STAT6(r=-0.664, r=-0.654, r=-0.629, r=-0.637, P<0.05), positive correlation with the expression levels of IL-13 (r=0.888, r=0.983, P<0.05).[Conclusion] Anchang Yuyang decoction can significantly promote intestinal mucosal repair in UC rats, the mechanism of which may be related to the regulation of the IL-13/JAK1/STAT6 signaling pathway and the up-regulation of the expression levels of Occludin and Claudin-1 in colon tissues.