目的 基于“亢害承制”理论探讨化瘀软肝胶囊干预肝纤维化模型大鼠细胞自噬的作用机制。方法 以SD雄性大鼠为研究对象，采用复合多因素方法（40% CCl₄花生油溶液皮下注射+高脂饲料+5%乙醇溶液饮水）复制肝纤维化大鼠模型8周，模型评价成功后依据随机数字表法分为正常组（NC）、模型组（Model）、秋水仙碱组（Col）、化瘀软肝胶囊低、中、高剂量组（HYRG-L、HYRG-M、HYRG-H），药物干预6周后处死，取血浆和肝组织，检测各组大鼠血清中ALT、AST，计算Liver Index水平，ELISA法检测IL-6含量的表达，天狼猩红染色观察肝组织胶原纤维沉积状况，Western Blot与免疫组织化学染色法检测α-SMA、LC3-B、Beclin-1的蛋白表达情况，RT-qPCR观察LC3-B、Beclin-1、α-SMA的基因水平变化。结果 与NC组比较，Model组大鼠肝脏组织中央静脉周围大量肝细胞脂肪变性、结缔组织增生、胶原纤维沉积，不同静脉间互相连接，形成纤维桥接，并可见炎性细胞浸润；Model组大鼠血清中ALT、AST及Liver Index水平增高（P<0.05），IL-6水平升高（P<0.05），肝脏组织中LC3-B、Beclin-1、α-SMA的蛋白与mRNA水平上升（P<0.05），与Model组相比较，Col组与化瘀软肝胶囊不同剂量组可不同程度调节ALT、AST及Liver Index水平（P<0.05，P<0.01），降低IL-6细胞因子水平，改善肝纤维化大鼠的纤维化程度，下调LC3-B、Beclin-1、α-SMA的蛋白与mRNA表达水平（P<0.05）。结论 中医学“亢害承制”理论指导下化瘀软肝胶囊抗肝纤维化的作用机制与其抑制炎症反应、阻碍HSC的过度增殖与活化、下调细胞自噬等途径有关。

Objective To investigate the mechanism of Huayu Rangan Capsule''s intervention on autophagy in liver fibrosis model rats based on the theory of "kanghai chengzhi"-a term of the five elements doctrine. It refers to the harm caused by excessive activity, which must be resisted and controlled.Methods The male SD rats were used as the research subjects to replicate the rat model of liver fibrosis by using a compound multi-factor method (subcutaneous injection of 40% CCl₄ peanut oil solution + high-fat feed +5% ethanol solution drinking water) for 8 weeks. After successful Model evaluation, the rats were divided into normal group (NC), Model group (Model), colchicine group (Col), Huayu Ruangan capsule low-dose, medium-dose and high-dose groups (HYRG-L, HYRG-M and HYRG-H) according to random number table method. After 6 weeks of drug intervention, the rats were sacrificed. Plasma and Liver tissue were collected, ALT AST in serum of rats in each group was detected, Liver Index level was calculated, IL-6 content was detected by ELISA, collagen fiber deposition in Liver tissue was observed by sirosin staining, α-SMA, LC3-B was detected by Western Blot and immunohistochemical stainingBeclin-1 protein expression in LC3-B and Beclin-1 α-SMA gene were observed by RT-qPCR.Results Compared with the NC group, a large number of hepatocyte steatosis, connective tissue hyperplasia and collagen fiber deposition around the central vein in the liver tissue of rats in the Model group were observed. Different veins were interconnected to form fibrous bridging, and inflammatory cell infiltration was also observed. The serum levels of ALT, AST and Liver Index in Model group were increased (P<0.05), the level of IL-6 was increased (P<0.05), the protein and mRNA levels of LC3-B, Beclin-1, α-SMA in Liver tissue were increased (P<0.05), compared with Model group, Col group and Huayu Ruangan Capsule groups with different doses could regulate the levels of ALT, AST and Liver Index to different degrees (P<0.05, P<0.01), reduce the level of IL-6 cytokines, and improve the degree of fibrosis in rats with Liver fibrosis. Down-regulation of LC3-B, Beclin-1, α-SMA protein and mRNA expression levels (P<0.05).Conclusions The mechanism of Huayu Ruangan Capsule against liver fibrosis under the guidance of the theory of "kanghai chengzhi" in traditional Chinese medicine is related to its inhibition of inflammatory response, blocking excessive proliferation and activation of HSC, and down-regulation of autophagy.

R285.5

甘肃省中医方药挖掘与创新转化重点实验室开放基金（zyzx-2020-02）：基于细胞自噬探讨化瘀软肝胶囊对HF大鼠Beclin-1及LC3-II表达的影响及其抗肝纤维化的机制，负责人：汪龙德；敦煌医学与转化省部共建教育部重点实验室开放基金（DHYX18-27）：化瘀软肝胶囊治疗气滞血瘀型非酒精性脂肪性肝病的临床疗效及对幽门螺旋杆菌影响的研究，负责人：汪龙德；甘肃省教育厅2021年优秀研究生“创新之星”项目资助（2021CXZX-766）：基于网络药理学从AKT/mTOR信号通路研究化瘀软肝胶囊干预HSC自噬的效应机制，负责人：牛媛媛。