摘要
目的:以四氯化碳(CCl4)诱导的肝纤维化大鼠为模型,探讨叶下珠对CCl4诱导的肝纤维化大鼠的保护作用及机制。方法:将60只SD大鼠随机分为正常组、模型组、秋水仙碱(0. 12 mg/kg)组和叶下珠(16 g/kg、8 g/kg、4 g/kg,生药量)组,每组10只。除正常组外,其余各组大鼠均腹腔注射40%CCl4花生油溶液(1 ml/kg),每周2次,连续8周,建立大鼠肝纤维化模型。从第一次注射CCl4花生油溶液开始,各治疗组分别灌胃给予相应药物,正常组和模型组给予等体积的生理盐水,每天1次,连续8周,8周末摘除眼球取血,收集肝脏。生化法检测血清中谷氨酸转氨酶(ALT)和门冬氨酸转氨酶(AST)活性;酶联免疫吸附试验(ELISA)测定血清中透明质酸(HA)、层黏连蛋白(LN)、Ⅲ型前胶原(PCⅢ)及Ⅳ型胶原(Ⅳ-C)表达水平;生化法检测肝组织中丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)水平; ELISA法检测肝组织中肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)和白介素-1β(IL-1β)含量;蛋白质免疫印迹法(Western Blot)测定肝组织中α-SMA、TGF-β1相对表达量;通过苏木精-伊红(HE)染色和Masson染色,显微镜下观察比较肝组织病理学变化。结果:与模型组比较,叶下珠各剂量组能明显降低大鼠血清中ALT、AST、LN、HA、PCⅢ和Ⅳ-C含量;并显著降低肝组织中MDA、TNF-α、IL-6及IL-1β水平,升高GSH-Px和SOD活性;下调肝组织纤维化相关蛋白TGF-β1、α-SMA表达;病理切片显示肝组织炎症坏死程度和肝纤维化病变程度明显改善。结论:叶下珠对CCl4诱导的肝纤维化大鼠具有明显保护作用,其机制可能与抑制氧化应激、炎症反应,调控α-SMA、TGF-β1蛋白表达有关。
Objective： To investigate the protective effect and mechanism of Phyllanthus urinaria on hepatic fibrosis in rats induced by carbon tetrachloride（CCl4）. Methods： Sixty SD rats were randomly divided into normal group,model group,colchicine group（0. 12 mg/kg） and high,medium and low dose of Phyllanthus urinaria group（16 g/kg,8 g/kg,4 g/kg）,with 10 rats in each group. Except the normal group,the other rats were injected intraperitoneally with 40% CCl4 peanut oil solution（1 ml/kg） to establish hepatic fibrosisa model,twice a week for 8 weeks. From the first injection of CCl4 peanut oil solution,each treatment group were given the corresponding drug intervention by gavage,normal group and model group were given equal volume of saline solution,once a day for 8 weeks. After 8 weeks,eyeballs were removed to collect blood,the livers were collected. Biochemical method was used to detect alanine aminotransferase（ALT） and aspartate aminotransferase（AST） in serum. The levels of serum HA,laminin（LN）,type Ⅲ（PCⅢ） and type Ⅳ collagen（Ⅳ-C） were detected by enzyme-linked immunosorbent assay（ELISA）. The levels of malondialdehyde（MDA）,superoxide dismutase（SOD） and glutathione peroxidase（GSH-Px） levels were detected by biochemical method. The content of TNF-α,IL-6 and IL-1β in liver tissues were detected by ELISA.Western Blot was used to determine the relative expression of α-SMA,TGF-β1 in liver tissues. The liver sections were prepared by hematoxylin-eosin staining（HE） and Masson staining. The pathological changes of liver tissue were observed under microscope. Results： Compared with the model group,the levels of ALT,AST,LN,HA,PCⅢ and Ⅳ-C in the serum were significantly decreased in high,medium and low dose group of Phyllanthus urinaria,the levels of MDA,TNF-α,IL-6 and IL-1β in liver tissue were significantly decreased and the activities of GSH-Px and SOD were increased. Liver tissue fibrosis related protein α-SMA,TGF-β1 expression was reduced. The path
作者
王刚
曹后康
曹秋妍
许琼梅
李跃龙
张可锋
Wang Gang;Cao Houkang;Cao Qiuyan;Xu Qiongmei;Li Yuelong;Zhang Kefeng(Guilin Medical University,Guilin 541004;Gansu University of Chinese Medicine,Lanzhou 730000)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2018年第4期104-108,共5页
Pharmacology and Clinics of Chinese Materia Medica
基金
广西八桂学者专项项目(桂财教函[2017]143号)
广西大学生创新创业训练计划项目(桂教高教[2017]4号)
关键词
叶下珠
肝纤维化
氧化应激
炎症反应
Phyllanthus urinaria
Hepatic fibrosis
Oxidative stress
Inflammatory response
