叶下珠及其复方抗乙肝病毒的临床及实验研究

【作者】 王荣国；

【导师】 王新华； 符林春；

【作者基本信息】 广州中医药大学 ， 中医临床基础， 2001， 硕士

【摘要】 叶下珠属植物在我国有着广泛的分布，具清肝、活血、解毒之效，治疗黄疸诸证有悠久历史。叶下珠复方是广州中医药大学热带医学研究所经过多年实验研究，临床观察，反复筛选，不断改进组成的具有清热解毒，疏肝健脾，活血化瘀功效，对慢性乙型肝炎有较好疗效的纯中药复方，由叶下珠、黄芪、三七等组成。 鸭乙肝体内模型仍是目前评价药物抗乙肝病毒作用的常用动物模型。但后天感染DHBV的鸭乙肝模型受毒种毒力影响大，缺乏稳定性，影响了对药物疗效的评价，需要建立一种更加稳定的鸭乙肝体内模型。斑点杂交法在鸭乙肝体内实验中应用较广，本实验进一步改进该实验操作条件，以期能更真实反映鸭外周血中DHBV-DNA水平变化。采用斑点杂交法直接检测广州麻鸭外周血中DHBV-DNA，以斑点出现为阳性，以0D490值作为DHBV-DNA水平，每10天采血1次，连续观察90天，结果广州麻鸭自然携带DHBV率为37.66％，外周血中DHBV-DNA水平高，维持时间长，适宜作为先天感染DHBV的实验动物，并以此建立起先天感染DHBV鸭乙肝模型。初步观察季节、饲料、DHBV阳性血清等对先天感染DHBV的广州麻鸭外周血中DHBV-DNA影响不大，说明该模型具有经济、简便、稳定性好的特点、是一种较理想的鸭乙肝模型。 本文观察了叶下珠及其复方等药物在先天感染DHBV鸭乙肝模型，后天感染DHBV的鸭乙肝模型中抗DHBV作用，结果海南叶下珠、福建叶下珠、广西叶下珠均具有一定作用；还观察到同一药物、相同剂量在先天感染DHBV模型中对DHBV-DNA的抑制作用较后天感染DHBV模型弱，这是否因先天感染DHBV的广州麻鸭外周血中DHBV-DNA水平高，维持时间长，且与先天感染DHBV麻鸭免疫耐受有关；观察到3TC、福建叶下珠单独使用具有较强抑制DHBV-DNA作用，联合使用不能增强抗DHBV作用，似乎与临床报道不一致；3TC对DHBV抑制作用不稳定，而ACV每次实验均能重复其对DHBV的抑制作用，是否与3TC稳定性差，易分解有关。 临床采用叶下珠复方联合抗乙肝特异性转移因子治疗40例慢性乙 型肝炎患者，并与单用叶下珠复方对照。结果治疗组总有效率92．5兄 HBeAe阴转率55．5％，HBV－DNA阴转率56．2％；对照组总有效串73．3＄， HBeAg阴转率44．0％，HBV－DNA阴转率43．4％，总有效串两组有显著性差 异（P＜0．05）；HBeAs、HBV－DNA阴转率两组无显著性差异（P＞0．05）。 总之，鸭乙肝先天感染粳型较后天励染模型具有稳定性好的优点。 叶下珠及其复方均有一定抑制DHBV－DNA作用，该抑制作用在先天感染 DHBV模型体内较后天感染模型体内低。叶下珠复方联合抗乙肝特异性转 移因子治疗慢性乙肝有增效作用。更多还原

【Abstract】 The traditional herb, Phyllanthus amarus(P.amaurs), distributes most parts of China, which has the effects of quenching evil heat in the liver, activating the blood and toxin-removing and has a long history of treatment many diseases, such as icterus. The compound of P.amaurs has been studied in lab, observed at hospital for many years and selected again and again and at last formed after many times improvement by Tropical Medicine Institute of Guangzhou University of TCM. Which is made up of P.amaurs, Astragalus membranaceus and Panax notoginseng, etc and is useful in the treatment of chronic hepatitis B virus infection and has the effects of clearing away heat and toxic materials, soothe the liver and invigorate the spleen and activating the blood circulation to eliminate blood stasis. DHBV model is an effective way to appraise the anti-HBV drug screening test at present. For being lack of stability and easily affected by toxin, the acquired infection DHBV ducks is not a good way to appraise the drug effect, so it is necessary to build a more stable and effective DHBV model. Spot-blot hybridization is widely used in this experiment. Some experiment conditions will be changed so that we can get the real data about the change of DHBV-DNA in peripheral blood of Guangzhou Brown spot ducks. Directly detect DHBV-DNA in peripheral blood of Guangzhou Brown spot ducks by spot-blot Hybridization method, view it as positive when spot appears, take 0D490 outcome as the level of DHB V-DNA, collect blood once every ten days, observe 90 days continually. The results showed that the natural infection rate of Guangzhou Brown spot Ducks was 37.66%, the level of DHBV-DNA was higher and lasted lOnger, so it is good as a congenital infection DHBV animal and on this basis we build congenital infection DHBV model. Through initial observation the season, food and DHBV positive serum do almost affect the level of DHBV-DNA in peripheral blood of Guangzhou Brown spot Ducks. This study suggested that this mode might be a good DHBV model for its economy, simplicity and good stability. This study observed the anti-DHBV effect of P.amaurs and its compound in Guangzhou Brown spot Ducks with congenital and acquired infection of DHBV. The results showed Hainan P.amaurs, Fujian P.amaurs and Guangxi P.amaurs are all useful, at the same time, also found the same drug, the same dosage, have the different effect, the inhibition tODHBV-DNA in acquired ducks is stronger than in congenital ducks. This maybe relative to high level DHBV-DNA, lasting longer time and the endurance of immunity. 3TC and Fujian P.amaurs used alone have a strong inhibition to DHBV-DNA, both used together did not show the additional effect of anti-DHBV, it appears contradict to clinic; 3TC’s inhibition to DHBV easily fluctuates, but ACV’s effect is stable. Now we are not quite sure whether it is relative to the bad stability and easy dissolution or not. We use the compound of P.amaurs and Anti Hepatitis B specific transfer factor together treating 40 chronic hepatitis B patients, and take the compound of P.amaurs alone as control class. The results showed the whole efficiency of the therapy group is 92.5%, HBeAg seroconversion is 55.5%, HBV-DNA seroconversion is 56.2%; the control class is 73.3%, HBeAg seroconversion is 44.4%, HBV-DNA seroconversion is 43.4%.更多还原