中南药学

目的 探讨痔瘘洗剂对痔疮模型大鼠的炎症反应、肛周组织病理学损伤的影响，明确痔瘘洗剂治疗痔疮的分子学机制。方法 70只SD雄性大鼠用随机数表法分为空白组，模型组，痔瘘洗剂低、中、高剂量组，分别给予315、621、1242 mg/kg，中药阳性药组，西药阳性药组，每组10只。使用99%浓度乙酸敷贴大鼠肛周构建大鼠痔疮溃烂模型。HE染色观察大鼠肛周组织病理形态，ELISA检测血清中白介素1β(IL-1β)、白介素6(IL-6)、白介素17A(IL-17A)、环氧酶-2(COX-2)、肿瘤坏死因子-α(TNF-α)分泌水平，RT-q PCR检测肛周组织HIF-1α、IL-17A、TNF-α、p65、PI3K、Akt、JAK2、STAT3 mRNA表达水平，Western blot检测肛周组织HIF-1α、IL-17A、TNF-α、p65、PI3K、Akt、JAK2、STAT3蛋白表达水平。结果 与模型组相比，痔瘘洗剂各剂量组、中药阳性药组、西药阳性药组均能改善大鼠肛周组织的溃疡，减少炎性细胞浸润，同时抑制血清中IL-1β、IL-6、IL-17A、COX-2、TNF-α的分泌水平，减少肛周组织中HIF-1α、IL-17A、TNF-α、p65、PI3K、Akt、JAK2、STAT3 mRNA表达，降低HIF-1α、IL-17A、TNF-α、p65、PI3K、Akt、JAK2、STAT3蛋白表达。结论 痔瘘洗剂对乙酸诱导的大鼠痔疮模型有明显的治疗作用，能够显著降低血清中炎症因子水平，并通过调控HIF-1α、IL-17A、TNF-α、p65、PI3K、Akt、JAK2、STAT3相关信号通路，降低HIF-1α、IL-17A、TNF-α、p65、PI3K、Akt、JAK2、STAT3蛋白表达，发挥清热解毒、消肿止痛、化湿止痒的作用。

Objective To evaluate the efficacy of hemorrhoid-fistula detergent(HFD) on inflammatory responses and histopathological injury in perianal tissues of a rat hemorrhoid model, and to explore its underlying molecular mechanisms of therapeutic action. Methods Seventy male Sprague-Dawley(SD) rats were randomly assigned using the random number table method into the following groups(10 rats per group): the normal group, the model group, HFD low, medium, and high dosage groups(315, 621, and 1242 mg/kg), the traditional Chinese medicine positive control group, and the Western medicine positive control group. A hemorrhoid ulcer model was established by topical application of 99% acetic acid to the perianal region. The pathological morphology of perianal tissue was observed by Hematoxylin-Eosin(HE) staining. The serum levels of interleukin-1β(IL-1β), interleukin-6(IL-6), interleukin-17A(IL-17A), cyclooxygenase-2(COX-2), and tumor necrosis factor-α(TNF-α) were quantified by enzyme-linked immunosorbent assay(ELISA). Reverse transcription-quantitative polymerase chain reaction(RT-qPCR) was employed to analyze the mRNA expression levels of hypoxia-inducible factor-1α(HIF-1α), IL-17A, TNF-α, p65(a subunit of NF-κB), phosphatidylinositol 3-kinase(PI3K), protein kinase B(Akt), Janus kinase 2(JAK2), and signal transducer and activator of transcription 3(STAT3) in perianal tissues. Western blot was conducted to determine the protein expression levels of HIF-1α, IL-17A, TNF-α, p65, PI3K, Akt, JAK2, and STAT3 in the same tissues. Results Compared with the model group, all dosage groups of HFD, as well as the TCM and Western medicine positive control groups, significantly ameliorated perianal tissue ulceration and reduced inflammatory cell infiltration; concurrently, they suppressed serum levels of IL-1β, IL-6, IL-17A, COX-2, and TNF-α, downregulated mRNA expression of HIF-1α, IL-17A, TNF-α, p65, PI3K, Akt, JAK2, and STAT3 in perianal tissues, and decreased protein expression of HIF-1α, IL-17A, TNF-α, p65, PI3K, Akt, JAK2, and STAT3. Conclusion HFD exhibited significant therapeutic effects on acetic acid-induced hemorrhoid model in rats. It markedly reduced the levels of inflammatory factors in serum and downregulated the protein expressions of HIF-1α, IL-17A, TNF-α, p65, PI3K, Akt, JAK2 and STAT3 via regulating the related signaling pathways involving these molecules, thereby exerting the effects of clearing heat and detoxifying, relieving swelling and pain, eliminating dampness and stopping itching.