Tang Qiao, Zhou Chao, Bai Zhaofang, Yao Qing, Chen Simin, Wen Xinru, He Zhaoyun, Zhang Jin, Li Ruisheng, Gong Man
Fifth Medical Center of Chinese PLA General Hospital, Beijing 100039, China.
School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Dec 20;44(12):2291-2299. doi: 10.12122/j.issn.1673-4254.2024.12.04.
To explore the role of the cGAS-STING signaling pathway in the therapeutic mechanism of Formula (LXJDHYF) for acute-on-chronic liver failure (ACLF) in mice.
Thirty C57BL/6 mice were randomly divided into blank control group, model group, low- and high-dose LXJDHYF groups, and H151 (a specific cGAS-STING pathway inhibitor) group (6). In all but the control group, the mice were treated with CCl to induce liver cirrhosis followed by intraperitoneal injections of lipopolysaccharide and D-amino galactose to establish mouse models of ACLF. After the treatments, the mouse livers were collected for HE and TUNEL staining, and serum levels of ALT, AST and TBil were determined. In bone marrow-derived macrophages (BMDMs) and liver tissues of ACLF mice, the expressions of cGAS-STING signaling pathway-related mRNAs including IFN‑β, ISG15, IL-6 and TNF-α were determined with RT-qPCR, and the phosphorylation levels of IRF3 and STING proteins were investigated using Western blotting.
Compared with the mice in the model group, the LXJDHYF-treated mice exhibited milder hepatocyte necrosis and inflammatory cell infiltration in the liver with significantly reduced hepatocyte apoptosis. LXJDHYF treatment also significantly lowered serum levels of ALT, AST, TBil, IL-6 and TNF-α in ACLF mice and effectively suppressed the expressions of cGAS-STING signaling pathway-related mRNA in both the BMDMs and the liver tissues and the phosphorylation of IRF3 and STING proteins in the BMDMs.
LXJDHYF can significantly improve liver function and attenuate inflammation in ACLF mice possibly by inhibiting excessive activation of the cGAS-STING signaling pathway.
探讨环鸟苷酸-腺苷酸合成酶-干扰素基因刺激蛋白（cGAS-STING）信号通路在方剂（LXJDHYF）对小鼠急性慢性肝衰竭（ACLF）治疗机制中的作用。
将30只C57BL/6小鼠随机分为空白对照组、模型组、低剂量和高剂量LXJDHYF组以及H151（一种cGAS-STING通路特异性抑制剂）组（每组6只）。除对照组外，其余小鼠均用四氯化碳处理以诱导肝硬化，随后腹腔注射脂多糖和D-氨基半乳糖以建立ACLF小鼠模型。处理后，收集小鼠肝脏进行苏木精-伊红（HE）和末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）染色，并测定血清谷丙转氨酶（ALT）、谷草转氨酶（AST）和总胆红素（TBil）水平。在ACLF小鼠的骨髓源性巨噬细胞（BMDM）和肝组织中，采用逆转录定量聚合酶链反应（RT-qPCR）测定cGAS-STING信号通路相关mRNA（包括干扰素-β（IFN-β）、干扰素刺激基因15（ISG15）、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α））的表达，并使用蛋白质免疫印迹法检测干扰素调节因子3（IRF3）和STING蛋白的磷酸化水平。
与模型组小鼠相比，LXJDHYF处理的小鼠肝脏中肝细胞坏死和炎性细胞浸润较轻，肝细胞凋亡明显减少。LXJDHYF处理还显著降低了ACLF小鼠血清中ALT、AST、TBil、IL-6和TNF-α的水平，并有效抑制了BMDM和肝组织中cGAS-STING信号通路相关mRNA的表达以及BMDM中IRF3和STING蛋白的磷酸化。
LXJDHYF可能通过抑制cGAS-STING信号通路的过度激活，显著改善ACLF小鼠的肝功能并减轻炎症反应。
