OBJECTIVE To investigate the mechanism of Yinzhi formula 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, high dose groups of Yinzhi formula, each consisting of six rats, and treated topically for six consecutive days. Additionally, the control group of six rats was set up. Perianal tissue lesion scores and ulcer areas were assessed on 2nd, 4th, 6th day. 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. 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 with the model group, the medium and high dose groups of Yinzhi formula significantly reduced perianal ulcer areas, tissue lesion scores, and pathological scores(P<0.05 or P<0.01); 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 Yinzhi formula 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.