银芷方调控JAK-STAT信号通路影响巨噬细胞极化改善痔疮大鼠肛周炎症

Yin Zhi Fang Modulates the JAK-STAT Signaling Pathway to Influence Macrophage Polarization for the Improvement of Perianal Inflammation in Rats with Hemorrhoids

关键词:  银芷方  痔疮  JAK-STAT信号通路  M1/M2巨噬细胞极化  炎症反应  

Key words:  yinzhi formula  hemorrhoids  JAK-STAT signaling pathway  M1/M2 macrophage polarization  inflammatory response  

摘要:

摘要：目的 基于JAK-STAT信号通路探讨银芷方干预M1/M2型巨噬细胞极化改善痔疮大鼠肛周炎症反应的作用机理。方法 采用醋酸贴敷法构建大鼠痔疮模型,造模成功后,分为模型组、马应龙组和银芷方低、中、高剂量组,每组6只,并另取6只正常大鼠为对照组。对药物干预组,连续6天在肛周病变区域涂抹对应药物,于2、4、6天进行肛周组织病变评分并计算溃疡面积；实验结束,取大鼠肛周组织HE染色观察大鼠肛周组织病理改变；免疫组化法检测大鼠肛周组织M1型巨噬细胞标志物CD86和M2型巨噬细胞标志物CD206的表达；ELISA法检测肛周组织中M1/M2型巨噬细胞极化因子水平；Western blot 法检测肛周组织中p-JAK2、p-STAT3、p-JAK1、p-STAT6及PPARγ蛋白表达变化。结果 与模型组比较,银芷方中、高剂量组肛周溃疡面积、组织病变及病理学评分均显著降低(P<0.05)；M1型巨噬细胞标志物CD86表达显著下调(P<0.05),M2型巨噬细胞标志物CD206表达显著上调(P<0.05)；细胞极化因子TNF-α水平降低(P<0.05)；IL-4、IL-10、TGF-β水平增加(P<0.05)；p-JAK1、PPARγ 蛋白水平显著上调(P<0.05或P<0.01),p-JAK2、p-STAT3蛋白水平显著下调(P<0.01或P<0.05)；此外,高剂量组还可明显降低IL-1β、COX2水平,并上调p-STAT6表达(P<0.05)。结论 银芷方可能通过调控JAK-STAT信号通路,干预巨噬细胞极化,减轻炎症反应,进而促进痔疮创面修复,改善痔疮症状。

Abstract:

ABSTRACT: OBJECTIVE To investigate the mechanism by which Yin Zhi Fang modulates the JAK-STAT signaling pathway to influence the polarization of M1/M2 macrophages, thereby improving the perianal inflammatory response in rats with hemorrhoids.METHODS A rat model of hemorrhoids was established using the acetic acid patch method. After successful modeling, the rats were divided into the model group, Ma Yinglong group, and low, medium, and high dose groups of Yin Zhi Fang, each consisting of six rats, and treated topically for six consecutive days. Additionally, a control group of six rats was set up. Perianal tissue lesion scores and ulcer areas were assessed on days 2, 4, and 6. Hematoxylin-eosin (HE) staining was used to observe pathological changes in perianal tissues. Immunohistochemistry was employed to detect the expression of the M1 macrophage marker CD86 and the M2 macrophage marker CD206. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of M1/M2 macrophage polarization factors in perianal tissues. Western blotting was performed to assess the expression changes of p-JAK2, p-STAT3, p-JAK1, p-STAT6, and PPARγ proteins in perianal tissues.RESULTS Compared to the model group, the medium and high dose groups of Yin Zhi Fang significantly reduced perianal ulcer areas, tissue lesion scores, and pathological scores (P < 0.05). The expression of the M1 macrophage marker CD86 was significantly downregulated (P < 0.05), while the expression of the M2 macrophage marker CD206 was significantly upregulated (P < 0.05). The level of the macrophage polarization factor TNF-α was decreased (P < 0.05), while IL-4, IL-10, and TGF-β levels were increased (P < 0.05). Protein levels of p-JAK1 and PPARγ were significantly upregulated (P < 0.05 or P < 0.01), and p-JAK2 and p-STAT3 protein levels were significantly downregulated (P < 0.01 or P < 0.05). Furthermore, the high dose group also significantly reduced IL-1β and COX2 levels and upregulated the expression of p-STAT6 (P < 0.05).CONCLUSION Yin Zhi Fang may alleviate inflammatory responses and promote hemorrhoid wound healing by modulating the JAK-STAT signaling pathway and intervening in macrophage polarization, thus improving hemorrhoid symptoms.