Department of Hepatology, School of Traditional Chinese Medicine, Capital Medical University, Beijing, People's Republic of China.
Department of Hepatology, Beijing Key Laboratory of Traditional Chinese Medicine Collateral Disease Theory Research, Capital Medical University, Beijing, People's Republic of China.
Drug Des Devel Ther. 2021 Sep 8;15:3845-3862. doi: 10.2147/DDDT.S308713. eCollection 2021.
Acute-on-chronic liver failure (ACLF) is a severe, complicated human disease. E2F1-mediated apoptosis plays an important role in ACLF development. Jieduan-Niwan (JDNW) formula, a traditional Chinese medicine (TCM), has shown remarkable clinical efficacy in ACLF treatment. However, the hepatoprotective mechanisms of the formula are barely understood.
This study aimed to investigate the mechanisms of JDNW formula in ACLF treatment by specifically regulating E2F1-mediated apoptotic signaling pathways in rats.
The JDNW components were determined by high-performance liquid chromatography (HPLC) analysis. The ACLF rat model was established using human serum albumin immune-induced liver cirrhosis, followed by D-galactosamine and lipopolysaccharide joint acute attacks. The ACLF rat was treated with JDNW formula. Prothrombin time activity was measured to investigate the coagulation function. Liver pathological injury was observed by hematoxylin-eosin (HE) and reticular fiber staining. The hepatocyte apoptosis index and apoptosis rate were determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and flow cytometry, respectively. Additionally, the expression of key genes and proteins that regulate E2F1-mediated apoptosis was analyzed by quantitative real-time PCR and Western blot.
Seven major components of JDNW formula were detected. The formula ameliorated the coagulation function, decreased the hepatocyte apoptosis index and apoptosis rate, and alleviated liver pathological damage in ACLF rats. The down-regulation of the expression of genes and proteins from p53-dependent and non-p53-dependent apoptosis pathways and the up-regulation of the expression of genes from blocking anti-apoptotic signaling pathways indicated that JDNW formula inhibited excessive hepatocyte apoptosis in ACLF rats via E2F1-mediated apoptosis signaling pathways.
The findings indicate that JDNW formula protects livers of ACLF rats by inhibiting E2F1-mediated apoptotic signaling pathways, implying that these pathways might be a potential therapeutic target for ACLF treatment.
急性肝衰竭（ACLF）是一种严重而复杂的人类疾病。E2F1 介导的细胞凋亡在 ACLF 的发展中起着重要作用。解毒软肝复方（JDNW）是一种中药方剂，已在 ACLF 的治疗中显示出显著的临床疗效。然而，该方剂的肝保护机制尚不清楚。
本研究旨在通过特异性调节 E2F1 介导的凋亡信号通路，探讨 JDNW 配方治疗 ACLF 的机制。
采用高效液相色谱法（HPLC）分析 JDNW 的成分。采用人血清白蛋白免疫诱导肝硬化，再用半乳糖胺和脂多糖联合急性攻击建立 ACLF 大鼠模型。用 JDNW 配方治疗 ACLF 大鼠。通过测定凝血酶原时间活动度来研究凝血功能。通过苏木精-伊红（HE）和网状纤维染色观察肝组织病理损伤。通过末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）法和流式细胞术分别测定肝细胞凋亡指数和凋亡率。此外，通过实时定量 PCR 和 Western blot 分析调节 E2F1 介导的凋亡的关键基因和蛋白的表达。
检测到 JDNW 配方的 7 种主要成分。该配方改善了 ACLF 大鼠的凝血功能，降低了肝细胞凋亡指数和凋亡率，减轻了肝组织病理损伤。p53 依赖性和非 p53 依赖性凋亡途径中基因和蛋白表达下调，阻断抗凋亡信号途径中的基因表达上调，表明 JDNW 配方通过 E2F1 介导的凋亡信号通路抑制 ACLF 大鼠过度的肝细胞凋亡。
这些发现表明，JDNW 配方通过抑制 E2F1 介导的凋亡信号通路来保护 ACLF 大鼠的肝脏，这表明这些通路可能是 ACLF 治疗的潜在治疗靶点。
