Tian Xiao-Ling, Zhang Yu, DU Shan, Wu Meng-Si, Tan Nian-Hua, Chen Bin
Hunan University of Chinese Medicine Changsha 410208, China the First Affiliated Hospital of Hunan University of Chinese Medicine Changsha 410007, China.
the First Affiliated Hospital of Hunan University of Chinese Medicine Changsha 410007, China.
Zhongguo Zhong Yao Za Zhi. 2024 Aug;49(15):4188-4196. doi: 10.19540/j.cnki.cjcmm.20240412.707.
Based on the hepatocyte growth factor(HGF)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt) signaling axis, this study investigated the therapeutic effect of Paeoniae Radix Rubra and Aconiti Lateralis Radix Praeparata(PRR-ALRP) her-bal pair on acute-on-chronic liver failure(ACLF) rats and its impact on hepatocellular regeneration. The rat model of ACLF was constructed by subcutaneous and tail vein injection of bovine serum albumin combined with intraperitoneal injection of lipopolysaccharides(LPS)+D-galactosamine(D-GalN). The rats were divided into normal control(NC) group, model(vehicle) group, PRR-ALRP(5.85 g·kg(-1)) group, and hepatocyte growth factor granules(HGFG, 4.05 g·kg(-1)) group. Hematoxylin-eosin(HE) staining was used to observe pathological changes in rat liver tissues. Serum alanine aminotransferase(ALT), aspartate transaminase(AST), and total bilirubin(TBIL) were detected using an automatic biochemical analyzer. The levels of interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) inflammatory factors were detected by ELISA. Immunofluorescence staining was used to detect the positive expression of proliferating cell nuclear antigen(PCNA), antigen identified by monoclonal antibody(Ki67), and cell cycle protein B1(CyclinB1). Real-time fluorescence-based quantitative polymerase chain reaction(RT-qPCR) and Western blot were used to detect the mRNA and protein expression levels of HGF, growth factor receptor-bound protein 1(Gab1), PI3K, Akt, phosphorylated PI3K(p-PI3K), and phosphorylated Akt(p-Akt). The results showed that compared with the vehicle group, the PRR-ALRP group had reduced liver tissue pathological scores, improved liver function, and reduced inflammatory response, with enhanced PCNA, Ki67, and CyclinB1 fluorescence expression. Furthermore, compared with the model group, the PRR-ALRP group showed upregulated expression of HGF and Gab1 proteins, as well as activation of PI3K and Akt phosphorylation. These findings suggest that PRR-ALRP herbal pair exerts anti-liver failure effects by alleviating hepatocyte inflammatory damage and promoting hepatocellular regeneration, and its specific regulatory mechanism may be related to the activation of the HGF/PI3K/Akt signaling pathway.
基于肝细胞生长因子（HGF）/磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）信号轴，本研究探讨了赤芍-制附子（PRR-ALRP）药对在慢性肝衰竭急性发作（ACLF）大鼠中的治疗作用及其对肝细胞再生的影响。通过皮下和尾静脉注射牛血清白蛋白联合腹腔注射脂多糖（LPS）+D-氨基半乳糖（D-GalN）构建ACLF大鼠模型。将大鼠分为正常对照（NC）组、模型（溶剂）组、PRR-ALRP（5.85 g·kg⁻¹）组和肝细胞生长因子颗粒（HGFG，4.05 g·kg⁻¹）组。采用苏木精-伊红（HE）染色观察大鼠肝组织病理变化。使用自动生化分析仪检测血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）和总胆红素（TBIL）。通过酶联免疫吸附测定（ELISA）检测白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）炎症因子水平。采用免疫荧光染色检测增殖细胞核抗原（PCNA）、单克隆抗体识别抗原（Ki67）和细胞周期蛋白B1（CyclinB1）的阳性表达。采用实时荧光定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测HGF、生长因子受体结合蛋白1（Gab1）、PI3K、Akt、磷酸化PI3K（p-PI3K）和磷酸化Akt（p-Akt）的mRNA和蛋白表达水平。结果表明，与溶剂组相比，PRR-ALRP组肝组织病理评分降低，肝功能改善，炎症反应减轻，PCNA、Ki67和CyclinB1荧光表达增强。此外，与模型组相比，PRR-ALRP组HGF和Gab1蛋白表达上调，PI3K和Akt磷酸化激活。这些研究结果表明，PRR-ALRP药对通过减轻肝细胞炎症损伤和促进肝细胞再生发挥抗肝衰竭作用，其具体调节机制可能与激活HGF/PI3K/Akt信号通路有关。
