本研究旨在探讨甘菊正丁醇提取部位(CGE)对四氯化碳(CCl_4)诱导的大鼠慢性肝损伤模型的缓解作用及机制。通过皮下注射CCl_4橄榄油溶液构建慢性肝损伤模型,CGE治疗4周后,血清天冬氨酸转氨酶(AST)水平,丙氨酸氨基转移酶(ALT),碱性磷酸酶(AKP),羟脯氨酸(HYP),白细胞介素-4(IL-4),白细胞介素-6(IL-6),丙二醛(MDA),超氧化物歧化酶(SOD),检测肿瘤坏死因子-α(TNF-α)。采用苏木素-伊红(HE)染色和Masson染色对大鼠肝脏组织进行处理,观察其结构。qPCR和Westernblot用于检测转化生长因子-β1(TGF-β1)/小母亲对无截瘫(Smad)的表达,Toll样受体4(TLR4),α-平滑肌肌动蛋白(α-SMA),和纤维连接蛋白(Fn)在大鼠肝组织和肝星状T6(HSC-T6)中的表达,并评估CGE对HSC活化的抑制作用。结果表明,CGE能显著降低血清AST水平,ALT,AKP,HYP,并影响IL-4、IL-6、TNF-α等相关炎症指标的水平,CCl_4诱导的大鼠慢性肝损伤中的MDA和MDA,对SOD活性没有影响,这可能会延迟肝损伤的过程,减轻肝脏胶原沉积和炎症浸润,对减轻大鼠慢性肝损伤有显著疗效。CGE可以抑制肝组织中α-SMA和TLR4蛋白的表达,逆转TGF-β1/Smad的表达,Fn,和TLR4在HSC-T6的体外表达。以上结果表明,CGE通过抑制HSC活化,减轻CCl_4诱导的大鼠慢性肝损伤,对大鼠肝组织有保护作用,并能改善大鼠肝组织的炎症反应和轻度肝纤维化。其药效机制可能与TGF-β1/Smad和TLR4相关表达有关。
This study aims to investigate the mitigating effect and mechanism of Cichorium glandulosum n-butanol extraction site(CGE) on the disease in carbon tetrachloride(CCl_4)-induced chronic liver injury model in rats. A chronic liver injury model was constructed by subcutaneous injection of CCl_4 olive oil solution, and after four weeks of CGE treatment, serum levels of aspartate aminotransferase(AST), alanine aminotransferase(ALT), alkaline phosphatase(AKP), hydroxyproline(HYP), interleukin-4(IL-4), interleukin-6(IL-6), malondialdehyde(MDA), superoxide dismutase(SOD), and tumor necrosis factor-α(TNF-α) were detected. Liver tissue was processed by hematoxylin-eosin(HE) staining and Masson staining to observe the structure of the rat liver. qPCR and Western blot were used to examine the expression of transforming growth factor-β1(TGF-β1)/small mothers against decapentaplegic(Smad), Toll-like receptor 4(TLR4), α-smooth muscle actin(α-SMA), and fibronectin(Fn) in rat liver tissue and hepatic stellate-T6(HSC-T6) and evaluate the inhibitory effect of CGE on HSC activation. The results showed that CGE could significantly reduce the serum levels of AST, ALT, AKP, HYP, and affect the levels of related inflammatory indexes including IL-4, IL-6, and TNF-α, and MDA in CCl_4-induced chronic liver injury in rats and had no effect on SOD activity, which could delay the process of liver injury, alleviate the hepatic collagen deposition and inflammatory infiltration, and had significant efficacy in mitigating chronic liver injury in rats. CGE could inhibit α-SMA and TLR4 protein expression in the liver tissue and reverse the increased TGF-β1/Smad, Fn, and TLR4-related expression in HSC-T6 in vitro. The above results indicated that CGE exerted hepatoprotective effects in rats by inhibiting HSC activation and alleviated CCl_4-induced chronic liver injury in rats and could ameliorate inflammatory response and slight liver fibrosis in rat liver tissue. Its pharmacodynamic mechanism might be related to TGF-β1/Smad and TLR4-related expression.