摘要
目的探讨在DEN诱导的大鼠肝癌模型中,护肝片在肝纤维化—肝癌发展过程中所产生的影响。方法雄性SD大鼠90只随机分为二乙基亚硝胺(diethylinitrosamine,DEN)组,正常对照组,护肝片组(921 mg·kg-1)。DEN组和护肝片组给予0.2%DEN溶液灌胃,按体重10 mg·kg-1给药,每周5次,至14周,对照组给予溶媒。护肝片组同时灌胃(ig)给予护肝片溶剂(羟甲基纤维素钠(CMC-Na)溶液配制),每日1次,直至16周。收集所有大鼠血液样本,进行血清学检测。通过病理组织学检查检测肝纤维化的程度。采用试剂盒检测肝组织中羟脯氨酸的含量。采用免疫组化法检测肝组织中α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)及胎盘型谷胱甘肽硫转移酶(Glutathione S-transferase placenta,GST-P)蛋白的表达。结果 DEN能够成功的诱导大鼠在12周发生肝纤维化,在16周发生肝癌。护肝片能够显著抑制肝纤维化—肝癌的发生。在12周时,护肝片能显著降低与肝功能相关的血清学指标以及α-SMA的过度表达,降低纤维化的程度。在16周时,护肝片能显著降低DEN诱导的大鼠肝癌的发生率和多样性。此外护肝片还能降低癌前病变指标γ-谷氨酰转肽酶(gamma-glutamyl transferase,GGT)和GST-P蛋白的表达。结论护肝片能够通过降低癌前病变指标GGT和GST-P的表达来阻止DEN诱导的肝癌的发生,同时也能减轻肝纤维化的程度,提示GGT与GST-P表达增强与肿瘤的发生密切相关,可能可以作为由纤维化进一步发展为肿瘤的标志,或者有可能成为预测肝癌发生的指标。
Objective To investigate the effects of liver-aid tablets on the development of hepatic fibrosis-carcinoma using a rat model of DEN-induced hepatocarcinogenesis. Methods The 90 rats were randomly assigned into three groups ： control group, DEN alone group, liver-aid tablets group（921 mg . kg^-1 ）. The rats in DEN alone and liver-aid tablets groups were treated with 0.2% DEN in CMC-Na by gavage 5 times a week for 14 weeks. During the experimental period, the rats in liver-aid tablets group were concomitantly administered liver-aid tablets in 0.5% CMC-Na for 16 weeks. Blood samples were collected from all rats for determination of serum enzymes levels. Histopathologic examination was carried out on all animals to determine the extent of fibrosis in rat liver and the development of HCC, and hydroxyproline content of the liver was also measured. In addition,the expression of α-smooth muscle actin（α-SMA） and glutathione S transferase placenta （GST-P） were detected by immunohistochemical method. Results DEN administration successfully induced hepato carcinogenesis in rats that caused liver fibrosis at 12th week and HCC at 16th week. However,liver-aid tablets significantly inhibited the development of hepatic fibrosis-carcinoma. At 12th week,liver-aid tablets significantly decreased the serum parameters of liver function, the extent of fibrosis and the overexpression of α-SMA. At 16th week, the incidence and multiplicity of development of HCC in DEN group were significantly decreased by liver-aid tablets. In addition, the GST-P and GGT positive foci, as a marker for neoplastic and preneoplas tic change ,were highly expressed in rat liver of DEN group while were obviously reduced by liver-aid tablets treatment. Conclusions During the process of DEN induced hepatic fibrosis-carcinoma in rats, the expressions of GGT and GST-P are inhibited by liver-aid tab lets, suggesting the injury mechanisms of DEN may involved GGT and GST-P.
出处
《安徽医药》
CAS
2013年第10期1652-1655,共4页
Anhui Medical and Pharmaceutical Journal
基金
国家自然科学基金资助项目(No 81073098)
