摘要
探究在四氯化碳(carbon tetrachloride,CCl_(4))诱导建立的大鼠慢性肝损伤模型中,毛菊苣正丁醇部位(Cichorium glandulosum n-butanol extraction site,CGE)对疾病的缓解作用及机制。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,检测大鼠血清中天冬氨酸氨基转移酶(aspartate aminotransferase,AST)、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、碱性磷酸酶(alkaline phosphatase,AKP)、羟脯氨酸(hydroxyproline,HYP)、白细胞介素-4(interleukin-4,IL-4)、白细胞介素-6(interleukin-6,IL-6)、丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)及肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平;并对肝组织进行苏木精-伊红(hematoxylin-eosin,HE)和马松(Masson)染色观察大鼠肝脏组织结构;采用qPCR、Western blot检测大鼠肝组织和肝星状细胞-T6(hepatic stellate-T6,HSC-T6)中转化生长因子-β1(transforming growth factor-β1,TGF-β1)/细胞信号传导分子(small mothers against decapentaplegic,Smad)、Toll样受体4(Toll-like receptor 4,TLR4)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)和纤维连接蛋白(fibronectin,Fn)的表达,评价CGE对HSC活化的抑制作用。结果显示,CGE能够明显减少CCl_(4)诱发的慢性肝损伤大鼠血清AST、ALT、AKP、HYP水平,并影响相关炎症指标IL-4、IL-6、TNF-α和MDA水平,对SOD活性无影响,可延缓肝损伤进程,缓解肝脏胶原沉积及炎症浸润,对大鼠慢性肝损伤缓解作用显著。CGE可以抑制肝脏组织中α-SMA、TLR4蛋白的表达,体外逆转HSC-T6中增加的TGF-β1/Smad、Fn和TLR4相关表达。上述结果表明,CGE可通过抑制HSC活化发挥对大鼠肝保护作用,缓解CCl_(4)诱导的大鼠慢性肝损伤,同时改善大鼠肝组织炎症反应及轻微肝纤维化,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.
作者
潘培妍
苏林洁
秦冬梅
PAN Pei-yan;SU Lin-jie;QIN Dong-mei(Key Laboratory of Xinjiang Phytomedicine Resource and Utilization of Ministry of Education/School of Pharmacy,Shihezi University,Shihezi 832002,China;Department of Pharmacy,Second Affiliated Hospital,Xinjiang Medical University,Urumqi 830063,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2024年第10期2566-2574,共9页
China Journal of Chinese Materia Medica
基金
国家自然科学基金地区项目(82160742)
新疆生产建设兵团重点领域科技攻关计划项目(2023CB008-25)。
