【作者】 杨静；

【导师】 江劲波； 邓常青；

【作者基本信息】 湖南中医药大学 ， 中医内科学， 2006， 硕士

【摘要】 目的：探讨补阳还五汤及其有效部位生物碱和苷对动脉血栓形成大鼠血浆TXA2.PGI2的影响及对大鼠血小板聚集和血小板内cAMP.cGMP的影响,从血小板活化的角度研究该方及其有效部位抗动脉血栓形成作用的机制,从而初步揭示该方抗动脉血栓形成的物质基础和抗血小板机理。方法：1.体内实验：将SD大鼠随机分为6组,假手术组,腹腔注射(ip)生理盐水10ml/kg；模型组,腹腔注射(ip)生理盐水10ml/kg；生物碱组,腹腔注射(ip)生物碱0.209g/kg；苷组,腹腔注射(ip)苷1.63g/kg；补阳还五汤原方组,腹腔注射(ip)原方7.5g/kg；抵克利得组,灌胃(ig)抵克利得0.03g/kg。实验前12小时给药一次,再于实验前给药一次,参照Kurz法改良制作大鼠颈总动脉血栓模型,于右侧颈总动脉插管取血,离心取上层贫血小板血浆,按放射免疫法测定血浆TXB2和6-keto-PGF1α含量。以上动物取血后剪下血栓部位血管,测定血栓重量。2.体外实验：将SD大鼠随机分为5组,即空白组、生物碱组、苷组、原方组、抵克利得组,剂量、给药时间、给药方式同体内实验。于左侧颈总动脉插管取血,离心取富血小板血浆(PRP),进行血小板计数及ADP诱导的血小板聚集实验。取血小板聚集试验前、后的PRP提取血小板cAMP、cGMP,采用放免法检测cAMP、cGMP。结果：1.模型组造模后,颈总动脉有明显的血栓形成。与模型组比较,补阳还五汤原方组、生物碱组、苷组和抵克利得组血栓重量均显著低于模型组(均为P<0.05)。2.与假手术组比,模型组血浆TXB2含量升高(P<0.05),血浆6-keto-PGF1α含量降低(P<0.05)。原方组血浆TXB2显著高于假手术组(P<0.05)而与模型组接近(P>0.05),但其血浆6-keto-PGF1α含量显著高于模型组(P<0.05)。生物碱组血浆TXB2含量较模型组显著降低(P<0.05),其血浆6-keto-PGF1α含量显著高于模型组(P<0.05)。苷组TXB2含量较模型组有所降低,但差异无显著性意义(P>0.05),其6-keto-PGF1α含量略低于假手术组而与模型组接近(均为P>0.05)。抵克利得组血浆TXB2和6-keto-PGF1α与模型组比较差异均无显著性意义(P>0.05)。3.各组血小板聚集比较,生物碱组血小板聚集强度与空白组相比显著降低(P<0.01)。苷组血小板聚集强度显著低于空白组(P<0.01)。原方组血小板聚集强度与空白组相比显著降低(P<0.05)。抵克利得组血小板聚集率显著低于空白组(P<0.01)。4.血小板cAMP比较,空白组血小板聚集后血小板内cAMP显著降低(P<0.05)。苷、原方组和抵克利得组聚集后血小板cAMP较聚集前显著降低(均为P<0.05)。生物碱组聚集后血小板cAMP虽较聚集前有所降低,但差异无显著性意义(P>0.05)。各组组间分析表明,聚集前后血小板cAMP降低值比较,各组差异均无显著性意义(P>0.05)。5.空白组聚集后血小板内cGMP降低(P<0.05),原方组聚集后血小板cGMP较聚集前显著降低(P<0.05)。而生物碱、苷和抵克利得组聚集后血小板cGMP虽有所降低,但差异无显著性意义(P>0.05)。各组组间分析表明,聚集前后血小板cGMP降低值比较,生物碱组cGMP降低值显著低于空白组(P<0.05)；苷、抵克利得和原方cGMP降低值与空白组比较,差异无显著性意义(P>0.05)。结论：补阳还五汤具有抗动脉血栓形成的作用,其有效部位生物碱和苷可能为其抗动脉血栓形成的主要物质基础,生物碱可通过促进PGI2产生,抑制血小板TXA2产生而实现抗动脉血栓的作用,原方的抗血栓作用与其促进PGI2产生有关,苷抗血栓作用可能不是通过调节TXA2和PGI2产生介导的。在对血小板聚集影响的研究中发现,各药物均可抑制ADP介导的血小板聚集,生物碱抗血小板聚集作用与其抑制血小板聚集时cAM P、cGM P降低有关,原方对血小板聚集后cAMP、cGM P的降低的抑制作用弱,苷抗血小板聚集作用不是通过增加血小板内cAM P、cGM P而实现的,可能与其它途径有关。结果提示生物碱类有效部位可能系该方抗血小板作用的主要物质,其作用与抑制TXA2产生,促进PGI2合成,增加血小板cAM P、cGM P有关。更多还原

【Abstract】 Objective:To explore effect of Buyang Huanwu Decoction (BHD)and its active fractions such as alkaloid and glycoside extracted from BHD on arterial thrombosis in rats and rat platelet aggregation and cAMP of the platelet and cGMP of the platelet. Research antithrombotic form mechanism of Buyang Huanwu Decoction (BHD)and its active fractions from the angle of platelet activation and reveal substantial foundation on antithrombotic form and mechanism of antiplatelet activation about BHD.Methods:1.Inbody experiment:Carotid thrombosis model in rats was induced by topical ferric chloride. The model rats were administered with BHD,alkaloid,glycoside and ticlopidine respectively. Weight of thrombosis was determined in each group.The levels of TXB2 and 6-keto-PGF1αin plasma in rats were measured by radioimmunoassay. 2.Outbody experiment:Divide SD rats into five groups,inclouding BHD group,alkaloid group,glycoside group and ticlopidine group.Drug doses,time and means of administering drugs are the same to experiment 1.Blood result from nearside carotid, PRP is abstracted by acentric means, number of platelets are acculated and experiment of platelet aggregation induced by ADP is carried out. The levels of cAMP and cGMP resulting from blood plasma in rats before and after platelet aggregation were measured by radioimmunoassay.Results:1.After model group made model, carotid had significant thrombus form.Weights of thrombosis in BHD group, alkaloid group, glycoside group and ticlopidine group were significantly lower than that of model group(p<0.05).2.The levels of TXB2 were increased and the levels of 6-keto-PGF1αwere decreased markedly in model group(p<0.05). The levels of TXB2 in BHD group were markedly higher than that of the sham-operated group (p<0.05),and approachable to that of the model group(p>0.05). BHD group could increase the levels of 6-keto-PGF1αwhen compared to the model group(p<0.05). Compared with the model group, alkaloid group could decrease the levels of TXB2 and increase the levels of 6-keto-PGF1αsignificantly(p<0.05). The tendency of the decreased TXB2 was noted in glycoside group, but there was no significant difference in TXB2 between glycoside group and the model group(p>0.05).The levels of 6-keto-PGF1αin glycoside group were lower than that of the sham-operated group(p<0.05), and approachable to that of the model group(p>0.05).3.After platelet aggregation inhibitations of BHD,alkaloid and glycoside groups were markedly lower than that of blank group (p<0.05).4.Compared with cAMP before platelet aggregation,cAMP of blank group,BHD group, glycoside group and Ticlopidine group after platelet aggregation could be decreased significantly(p<0.05).The tendency of the decreased cAMP after platelet aggregation was noted in alkaloid group, but there was no significant difference before platelet aggregation (p>0.05).Compared with decreasion of cAMP in every group before and after platelet aggregation, there was no significant difference (p>0.05).5. cGMP of blank group and BHD group after platelet aggregation could be decreased significantly(p<0.05).The tendency of the decreased cGMP after platelet aggregation was noted in alkaloid group,glycoside group and Ticlopidine group, but there was no significant difference before platelet aggregation (p>0.05).Compared with decreasion of cGMP in every group before and after platelet aggregation, cGMP of alkaloid group after platelet aggregation could be decreased significantly than that of blank group (p<0.05). when compared to the blank group,there was no significant difference in glycoside group,BHD group and Ticlopidine group (p>0.05).Conclusions:BHD had effections of Antithrombotic form.Maybe alkaloid and glycoside are main substantial foundation on antithrombotic form. The efficacy of alkaloid may contribute to inhibiting production of TXA2 and increasing synthesis of PGI2. Antithrombotic action of glycoside was not based on its modulation on TXA2 and PGI2,it may be concerned with other aspects.In resarch of effection of platelet aggregation,every medicine could inhabit platelet aggregation by ADP. Effection of antiplatelet aggregation of alkaloid was relative to inhabition of cAMP and cGMP when platelet was aggregated. Antiplatelet aggregation action of glycoside was not based on increasement of cAMP and cGMP in platelets,it may be concerned with other aspects.Result reveals that alkaloid may be main substantial foundation on antiplatelet action and may be relative to inhibiting production of TXA2, producing synthesis of PGI2 and increasing cAMP and cGMP in platelets.更多还原