复方小花清风藤浸膏抗急性肝损伤作用及对炎症干预作用的机制研究

【作者】 杨龙飞；

【导师】 孙建宁；

【作者基本信息】 北京中医药大学 ， 中西医结合基础， 2006， 博士

【摘要】 很多因素都可以引起肝脏损伤,包括外伤导致的肝损伤、化学性肝损伤(包括药物性肝损伤和非药物性肝损伤,主要是由于平时用药及酒精等因素造成的肝脏损伤)以及病毒性肝炎所导致的肝损伤等。其中以病毒性肝炎引起的肝损伤最为常见。我国是HBV感染的高发区,根据1992-1995年全国病毒性肝炎血清流行病学调查,人群HBsAg携带率达9.75%,约1.2亿人。探讨中药对肝脏的保护作用以及抗炎的机制对开发新的抗肝炎药物有重要意义。近年的研究表明:肝炎的发生机制除与引起炎症的病毒有关外,还与机体的免疫机制引起的肝脏损伤有关。炎症的组织学特征是炎性细胞浸润,细胞因子、炎症介质和转录因子等都在肝炎发病的病理过程中起关键性作用。小花清风藤(Sabia. parviflora Wall. ex Roxb)为苗族民间药物,是清风藤科清风藤属藤本植物,主要分布于云南、广西、贵州等地,以前的研究资料表明,其对甲肝及乙肝的治疗效果均较显著,有很好的利湿退黄和降转氨酶作用。但相关的药理研究资料极少。贵州省科技产业厅立项,对该药进行化学和活性的系统研究,并研制抗肝损伤的新药。复方小花清风藤浸膏主要由小花清风藤、三七、石斛组成,有清肝利胆的功效,主要用于治疗病毒性肝炎。本研究采用四氯化碳、α-萘异硫氰酸酯、D-氨基半乳糖、卡介苗联合脂多糖诱发四种肝损伤模型,采用生物化学、酶联免疫分析、免疫组织化学、逆转录聚合酶链反应等手段,从肝脏功能、脂质过氧化、细胞因子的表达、肝细胞凋亡以及转录因子的活化等环节探讨了复方小花清风藤浸膏对不同肝损伤模型的保护作用及其对炎症干预作用的相关机制。第一部分复方小花清风藤对急性肝损伤动物模型的保护作用1复方小花清风藤对四氯化碳(CCL4)、α-萘异硫氰酸酯(ANIT)致小鼠肝损伤模型的保护作用目的:研究复方小花清风藤对四氯化碳致急性化学性肝损伤模型及α-萘异硫氰酸酯致胆瘀症模型小鼠血清中转氨酶、碱性磷酸酶及胆红素含量的影响,探讨复方小花清风藤对肝细胞膜通透性和胆汁排泄功能的影响。方法:分别采用四氯化碳皮下注射、α-萘异硫氰酸酯灌胃致小鼠急性肝脏损伤,观察复方小花清风藤对四氯化碳致肝损伤模型小鼠血清中谷丙转氨酶(ALT)及谷草转氨酶(AST)含量的影响;α-萘异硫氰酸酯致肝损伤模型小鼠血清中ALT、AST、碱性磷酸酶(ALP)及总胆红素(T-Bil)含量的影响。结果:两种模型均可见小鼠血清中的转氨酶含量显著升高,α-萘异硫氰酸酯灌胃48小时后,小鼠血清中的ALP及T-Bil显著升高。复方小花清风藤6.5g/kg、3.3g/kg可明显降低四氯化碳致肝损伤模型小鼠血清中的ALT含量,13g/kg、6.5g/kg、更多还原

【Abstract】 Liver injury can be caused by many factors, including trauma, chemical materials (mostly drugs or alcohol) and hepatic virus, etc. The most common cause is hepatitis virus. HBV infection rate is high in our country. According to the epidemiologic investigation on hepatitis serum in 1992 to 1995, the HBsAg rate arrive 9.75%, this means about 1.2 hundred million people have HbsAg in serum. So it is significant to investigate the protection effect of traditional Chinese medicine and the anti-inflammation mechanism. In recent years, it is proved that hepatitis is related with not only virus, but also the body immune system. The histological sign of inflammatory include leukocyte infiltration, cytokines, inflammatory mediums and transcription factors, etc.Sabia. parviflora Wall. ex Roxb mainly grow in Yunnan, Guangxi, Guizhou province. The research materials in before show that it has perfect effect on hepatitis A and hepatitis B. The effect of decreasing ALT, AST and T-Bil is even better. But the concerned pharmacological materials are quite few. In this paper, Parviflora compond was mainly composed of Sabia. parviflora Wall. ex Roxb. By the means of biochemical assay, enzyme-linked immunosorbent assay (ELISA), immunohistochemistry (IHC) and molecular biology, effects of Parviflora compond on the inflammatory reaction in acute liver injury models were studied from liver tissue damage, leukocyte infiltrate, cytokine expression and transcription factors activation, etc. The chief results are as follows.Part One : Protective effects of Parviflora compond on acute liver injury models1 Protective effects of Parviflora compond on liver injury models induced by CCL4 and ANIT in miceOBJECTIVE: To study the effects of Parviflora compond on serum ALT, AST, ALP and T-Bil in liver injury models im mice induced by CCL4 and ANIT. METHODS:The two acute liver injury models were established by CCL4 ip and ANIT ig. Serum ALT, AST, ALP, T-Bil was determined. RESULTS:Serum ALT and AST increased in both models. ANIT ig 48 hours later, serum ALP and T-Bil obviously increased. Parviflora compond (6.5g/kg, 3.3g/kg, ig) can significantly decrease serum ALT level in mice of CCl4 liver injury model. Parviflora compond (13g/kg, 6.5g/kg, 3.3g/kg, ig) can decrease serum ALP and T-Bil level in mice of ANIT liver injury model. CONCLUSION:Parviflora compond have更多还原