强力宁对四氯化碳、乙醇诱导的肝纤维化大鼠模型肝脏Ⅰ、Ⅲ型前胶原mRNA表达的影响

Effects of Potenlini on mRNA Expression of Procollagen Type Ⅰ,Ⅲ in CCl 4 Plus Ethanol Induced Liver Injury and Hepatic Fibrosis in Rats

目的：探讨强力宁治疗对毒素性肝损伤－纤维化大鼠模型肝脏内Ⅰ、Ⅲ型前胶原ｍＲＮＡ表达的影响，从分子水平揭示强力宁对肝纤维化的治疗作用及机理。方法：采用四氯化碳皮下注射和饮用乙醇水溶液方法诱导大鼠肝损伤－纤维化模型，同时部分予以强力宁治疗，在四氨化碳等处理后定期处死大鼠收集血清和肝脏标本，采用异硫氰酸胍－酚－氯仿一步法抽提肝组织总ＲＮＡ，用地高辛标记的Ⅰ、Ⅲ型前胶原ｃＤＮＡ探针与各组肝组织总ＲＮＡ进行斑点杂交，比较各组标本肝脏Ⅰ、Ⅲ型前胶原的表达水平。结果：四氯化碳等损伤后肝脏内Ⅰ、Ⅲ型前胶原ｍＲＮＡ表达显著增高，而强力宁能够抑制损伤后肝脏内Ⅰ、Ⅲ型前胶原ｍＲＮＡ的异常增高的表达。结论：强力宁能够通过抑制毒素致肝损伤动物模型肝脏内Ⅰ、Ⅲ型前胶原基因的表达来抑制肝纤维化的发生。

Abstract Aims: In these study,the effects of Potenlini on procollagen type Ⅰ,Ⅲ mRNA expression in cirrhostic livers of rats had been studied in order to clarify the mechanism and effects of potenlini in molecular level. Material and methods:Rat model of liver injury and hepatic fibrosis were obtained by subcutaneous injections of CC1 4 twice weekly and administration of 10% ethanol in drinking water.Normal control group,Potenlini treatment group and normal control group were established at the same time.By the weeks of 2,6 and 9 after CC1 4treatment,the rats in each group were killed randomlly,liver samples were collected.HE stain and collagen special stain were used to observe the histological changes in livers.The total RNA of liver tissue was extracted by single step RNA isolation method (Acid Guanidinium Thiocyanate Phenol Chloroform Extraction).DIG labeled cDNA probes of type Ⅰ,Ⅲ procollagen and collagenase were used to hybridize with dot blotted total liver RNA samples.The conclusions were photographed and analysed by computer and changed into data which could be analysed and compared statistically.Results:According to microscopic observation,in model control group,at the second week the main pathologic