目的 基于调控内源性逆转录病毒（endogenous retroviruses，ERVs）/维甲酸诱导基因I样受体（retinoic acid-inducible gene I-like receptors，RLRs）/I型干扰素（type I interferon，IFN-I）信号通路，探讨黄芪-丹参药对（Astragali Radix-Salviae Miltiorrhizae Radix et Rhizoma herb pair，HQ-DS）治疗肝纤维化（hepatic fibrosis，HF）的作用机制。方法 通过超高效液相色谱-串联质谱（ultra-performance liquid chromatography-tandem mass spectrometry，UPLC-MS/MS）对HQ-DS提取物以及含药血清进行成分分析与鉴定。将48只雄性C57BL/6J小鼠随机分为对照组、模型组及HQ-DS低、中、高剂量（4.55、9.10、18.20 g/kg）组和水飞蓟素（100 mg/kg）组，每组8只。采用四氯化碳（carbon tetrachloride，CCl₄）建立HF小鼠模型，给予药物干预后，检测血清中肝功能相关指标；通过苏木素-伊红（hematoxylin-eosin，HE）染色、Masson和天狼星红染色观察肝组织病理变化；流式细胞术检测肝组织巨噬细胞浸润情况；Western blotting检测肝组织纤维化相关蛋白表达；采用高通量转录组测序分析肝组织基因转录水平变化；通过qRT-PCR、Western blotting和ELISA检测肝组织ERVs/RLRs/IFN-I信号通路相关因子表达。通过Mavs^-/-小鼠建立HF模型后给予HQ-DS进行干预，验证HQ-DS抗HF的作用机制。体外实验采用CCK-8实验评估HQ-DS对脂多糖（lipopolysaccharide，LPS）、尼日利亚菌素（Nigericin）刺激iBMDM细胞活力的影响，免疫荧光检测dsRNA表达，qRT-PCR和Western blotting进一步验证HQ-DS对ERVs激活的抑制作用。结果 与对照组比较，模型组小鼠体质量增加缓慢，肝组织出现空泡样变性以及炎症细胞浸润，血清中肝功能相关指标显著升高（P＜0.001），肝组织中ERVs/RLRs/IFN-I信号通路相关因子表达显著升高（P＜0.001）；与模型组比较，HQ-DS和水飞蓟素干预后小鼠体质量增长缓慢显著改善（P＜0.01、0.001），肝组织空泡变性及炎症浸润程度明显减轻，血清中肝功能相关指标显著降低（P＜0.01、0.001），HF进程得到有效缓解，肝组织ERVs/RLRs/IFN-I信号通路相关因子表达显著降低（P＜0.05、0.01、0.001）。进一步的机制验证显示，在Mavs^-/-小鼠中，CCl₄诱导的促炎因子升高被显著抑制（P＜0.001），且ERVs/RLRs/IFN-I通路相关因子的mRNA表达水平亦被显著下调（P<0.001）；给予HQ-DS后炎症因子和ERVs/RLRs/IFN-I通路相关因子的mRNA表达水平均无显著改变。细胞实验中，与模型组比较，HQ-DS显著提高iBMDM细胞活力（P＜0.001），减少dsRNA积累，并下调ERVs及IFN-I通路核心基因的表达（P＜0.05、0.01、0.001），同时抑制RLRs通路关键蛋白磷酸化TANK结合激酶1（phosphorylated TANK-binding kinase 1，p-TBK1）、磷酸化干扰素调节因子3（phosphorylated interferon regulatory factor 3，p-IRF3）的表达（P＜0.05、0.01、0.001）。结论 HQ-DS通过抑制ERVs/RLRs/IFN-I信号通路，减轻肝组织炎症损伤，缓解小鼠HF进程。

Objective To investigate the mechanism by which Huangqi (Astragali Radix)-Danshen (Salviae Miltiorrhizae Radix et Rhizoma) herb pair (HQ-DS) in treating hepatic fibrosis (HF) via regulating the endogenous retroviruses (ERVs)/retinoic acid-inducible gene I-like receptors (RLRs)/type I interferon (IFN-I) signaling pathway. Methods The components of HQ-DS extract and drug-containing serum were analyzed and identified using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). A total of 48 male C57BL/6J mice were randomly divided into control group, model group, HQ-DS low-, medium-, high-dose (4.55, 9.10, 18.20 g/kg) groups and silymarin (100 mg/kg) group, with eight mice in each group. A mouse model of HF was established using carbon tetrachloride (CCl₄), and after drug intervention, liver function related indicators in serum were detected. The pathological changes of liver tissue through hematoxylin eosin (HE) staining, Masson and Sirius red staining were observed. Flow cytometry was used to detect macrophage infiltration in liver tissue. Western blotting was used to detect the expressions of fibrosis related proteins in liver tissue. High-throughput transcriptome sequencing was used to analyze changes in gene transcription levels in liver tissue. The expressions of factors related to the ERVs/RLRs/IFN-I signaling pathway in liver tissue were detected using qRT-PCR, Western blotting and ELISA. A HF model was established in Mavs^-/- mice and intervened with HQ-DS to verify the mechanism of HQ-DS’s anti-HF effect. In vitro experiments were conducted using CCK-8 assay to evaluate the effect of HQ-DS on the activity of iBMDM cells stimulated by lipopolysaccharides (LPS) and nigericin. Immunofluorescence detection of dsRNA expression was performed, qRT-PCR and Western blotting were used to further verify the inhibitory effect of HQ-DS on the activation of ERVs. Results Compared with control group, the body weight of mice in model group was increased slowly, with vacuolar degeneration and inflammatory cell infiltration in liver tissue, liver function related indicators in serum were significantly increased (P < 0.001), and the expressions of factors related to the ERVs/RLRs/IFN-I signaling pathway in liver tissue were significantly increased (P < 0.001). Compared with model group, HQ-DS and silymarin intervention significantly improved the slow growth of body weight in mice (P < 0.01, 0.001), significantly reduced the degree of vacuolar degeneration and inflammatory infiltration in liver tissue, significantly reduced liver function related indicators in serum (P < 0.01, 0.001), effectively alleviated the process of HF, and significantly reduced the expressions of factors related to the ERVs/RLRs/IFN-I signaling pathway in liver tissue (P < 0.05, 0.01, 0.001). Subsequent verification of the underlying mechanisms revealed that in Mavs^-/- mice, the elevation of pro-inflammatory factors induced by CCl₄ was markedly suppressed (P < 0.001), the mRNA expression levels of factors associated with the ERVs/RLRs/IFN-I signaling pathway were significantly downregulated (P < 0.001). However, following the administration of HQ-DS, there was no significant change in the mRNA expression levels of inflammatory factors and factors related to the ERVs/RLRs/IFN-I pathway. In cell experiments, compared with model group, HQ-DS significantly increased iBMDM cell viability (P < 0.001), reduced dsRNA accumulation, and downregulated the expressions of core genes in ERVs and IFN-I pathways (P < 0.05, 0.01, 0.001), while inhibited the expressions of key proteins [phosphorylated TANK binding kinase 1 (p-TBK1), phosphorylated interferon regulatory factor 3 (p-IRF3)] in RLRs pathway (P < 0.05, 0.01, 0.001). Conclusion HQ-DS alleviates inflammatory injury and ameliorates the progression of HF in mice by inhibiting ERVs/RLRs/IFN-I signaling pathway.

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国家自然科学基金青年项目（82505414）；湖南省自然科学基金青年科学基金项目（2026JJ40093）；中药粉体与创新药物省部共建国家重点实验室培育基地开放基金（25PTKF1005，24PTKF1005）