苗药了哥王不同部位提取物的肾毒性研究

目的研究苗药了哥王不同部位提取物对健康大鼠的肾毒性作用,为了哥王的毒性作用机制及临床用药提供参考。方法以70%乙醇为溶剂,采用渗漉法提取,得了哥王乙醇总提取物;将上述提取物用水分散后,依次用石油醚、乙酸乙酯、正丁醇萃取得相应部位提取物,剩余为水部位提取物。将SD大鼠随机分为乙醇总提取物组、石油醚部位组、乙酸乙酯部位组、正丁醇部位组、水部位组和空白组,每组12只(雌雄各半)。给药组大鼠灌胃相应剂量的药液(乙醇总提取物317.520 mg/kg、石油醚部位7.875 mg/kg、乙酸乙酯部位78.435 mg/kg、正丁醇部位53.865 mg/kg、水部位76.545 mg/kg),每天1次,连续给药2周,再停药恢复2周;空白组大鼠灌胃等体积1.0%羧甲基纤维素钠溶液。实验期间观察大鼠的一般情况,取尿液(第14、28天)、血清和双肾组织(第15、29天),计算其肾脏指数,检测血清和尿液中的肾功能指标水平并观察肾组织的病理形态学变化。结果给药期间,与空白组比较,乙醇总提取物组、乙酸乙酯部位组大鼠出现精神萎靡、活动量和饮食量减少、大便稀薄、体质量下降等中毒行为活动特征;石油醚部位组、正丁醇部位组和水部位组大鼠精神状态较空白组稍差,活动量和饮食量稍减少,大便稀薄,体质量增长缓慢;但各组大鼠肛温无较大变化。给药2周后,乙醇总提取物组大鼠的肾脏指数,乙醇总提取物组、乙酸乙酯部位组大鼠血清中N-乙酰氨基葡萄糖苷酶(NAG)、尿素氮(BUN)、肌酐(Cr)水平,正丁醇部位组大鼠血清中NAG水平,水部位组大鼠血清中Cr水平,以及乙醇总提取物组、石油醚部位组大鼠尿液中NAG水平,乙酸乙酯部位组大鼠尿液中NAG、尿蛋白水平均显著升高(P<0.05或P<0.01);在病理形态学观察中,乙醇总提取物组、石油醚部位组和乙酸乙酯部位组大鼠有不同程度的肾小管结构不清、细胞肿胀、少数细胞坏死,伴有肾小球固缩、肾小管硬化、炎症细胞浸润。停药后,大鼠以上症状逐渐好转,精神状态有明显改善,活动量和饮食量增加,大便趋于正常。停药恢复2周后,各给药组大鼠血清和尿液中上述指标水平均恢复至与空白组接近(P>0.05);各给药组大鼠的肾小球结构逐渐恢复清晰,乙醇总提取物组、石油醚部位组和乙酸乙酯部位组大鼠少见细胞肿胀和炎症细胞浸润。结论苗药了哥王乙醇总提取物、石油醚部位和乙酸乙酯部位均具有一定的肾毒性,且具有一定的可逆性。其毒性成分可能为脂溶性成分。

Determination of the Content of Total Alkaloids in Different Solvent Extracts from Climbing Groundsel Herb Produced in Guizhou Province and Their Antibacterial Activity

[Objective] This study aimed to compare the contents of total alkaloids in different solvent extracts from climbing groundsel herb produced in Guizhou Province and their antibacterial activity. [Method] The content of total alkaloids was measured by UV spectrophotometry, and Oxford cups were used to investigate the antibacterial activity of each solvent extract. [Result] There was a good linear correlation between the absorbance measured by the UV spectrophotometer and the content of total al- kaloids within the concentration of 0.011 0-0.054 8 mg/ml at 207 nm, and the re- gression equation was Y=23.654X＋0.021, R=0. 999 7 and the average recovery rate was 99.2%. The contents of total alkaloids in 60% ethanol extract, 95% ethanol and water ethanol were 38.71, 52.25 and 60.50 mg/g, respectively. The 60% ethanol ex- tract had strong antibacterial activity against Streptococcus pneumoniae, Staphylo- coccus aureus and Escherichia coil; the water extractive had stronger antibacterial activity against Escherichia colr, 95% ethanol extract had weak antibacterial activity, with no inhibition of Pseudomonas aeruginosa. [Conclusion] This method is accurate, simple, with good repeatability, which can be used for the determination of alkaloids content of the climbing groundsel herb; there is no positive correlation between the content of total alkaloids in different solvent extracts of climbing groundsel herb and their antibacterial activity.