摘要
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。
OBJECTIVE To study the nephrotoxicity of the extracts from different parts of Miao medicine Wikstroemia indica in healthy rats,and to provide reference for the study of its toxicity mechanism and clinical drug use.METHODS Using 70%ethanol as solvent,total ethanol extract of W.indica was extracted with diacolation method.After dispersing the above extract with water,the fractions of corresponding fractions were obtained with petroleum ether,ethyl acetate and n-butanol,and the rest was the extract of water fraction.SD rats were randomly divided into total ethanol extract group,petroleum ether fraction group,ethyl acetate fraction group,n-butanol fraction group,water fraction group and blank group,with 12 rats in each group(half male and half female).The rats in the administration groups were given the corresponding dose of drug solution intragastrically(total ethanol extract 317.520 mg/kg,petroleum ether fraction 7.875 mg/kg,ethyl acetate fraction 78.435 mg/kg,n-butanol fraction 53.865 mg/kg and water fraction 76.545 mg/kg),once a day,for consecutive 2 weeks,and then stopped taking drug for 2 weeks;rats in the blank group were given equal volume of 1.0%sodium carboxymethyl cellulose solution intragastrically.During the experiment,the general conditions of rats were observed.The samples of urine(on the 14th and 28th day),serum and bilateral renal tissues(on the 15th and 29th day)were taken respectively,the renal index was calculated,the levels of renal function indexes in serum and urine were detected,and the pathomorphological changes of renal tissues were observed.RESULTS During administration,compared with blank group,the rats in the total ethanol extract group and ethyl acetate fraction group showed poisoning behavior and activity characteristics such as mental depression,decreased activity and diet,thin stool and decreased body mass.The mental state of the rats in the petroleum ether fraction group,n-butanol fraction group and water fraction group were slightly worse than that in blank group,and slightly decreased activity and diet as well as thin stool,and slowly increased body mass were found;however,there was no significant difference in anal temperature in each group.After 2 weeks of administration,the renal index in total ethanol extract group,the serum levels of N-acetylglucosaminidase(NAG),urea nitrogen(BUN)and creatinine(Cr)in total ethanol extract group and ethyl acetate fraction group,serum level of NAG in n-butanol fraction group and serum level of Cr in water fraction group,as while as NAG levels in urine of rats in total ethanol extract group and petroleum ether fraction group,NAG and urinary protein levels in urine of rats in ethyl acetate fraction group were increased significantly(P<0.05 or P<0.01).In the pathomorphological observation,renal tubules showed different degrees of unclear structure,cell swelling and a few cell necrosis in the total ethanol extract group,petroleum ether fraction group and ethyl acetate fraction group,accompanying by glomerular pyknosis,renal tubular sclerosis and inflammatory cell infiltration,compared with blank group.After drug withdrawal,the mental state of rats in the administration groups were significantly improved,the amount of activity and diet increased,and the stool tended to be normal.Two weeks after drug withdrawal and recovery,the levels of above indexes in serum and urine of rats in administration groups returned to be close to that in blank group(P>0.05);the glomerular structure of rats in each administration group gradually recovered clearly,and cell swelling and inflammatory cell infiltration were rare in total ethanol extract group,petroleum ether fraction group and ethyl acetate fraction group.CONCLUSIONS The total ethanol extract,petroleum ether fraction and ethyl acetate fraction of Miao medicine W.indica have certain nephrotoxicity and reversibility.The toxic component may be fat soluble.
作者
任晨晨
冯果
李玮
王建科
彭礼珍
朱光林
王文静
苏红梅
REN Chenchen;FENG Guo;LI Wei;WANG Jianke;PENG Lizhen;ZHU Guanglin;WANG Wenjing;SU Hongmei(School of Pharmacy,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;National Engineering Research Center of Miao Medicine,Guiyang 550025,China)
出处
《中国药房》
CAS
北大核心
2022年第4期419-424,共6页
China Pharmacy
基金
国家自然科学基金资助项目(No.81760766,No.8206-0767)
全国中医药创新骨干人才培训项目(No.国中医药人教函[2019]128号)
贵州省药物新剂型新工艺科技创新人才团队项目(No.黔科合平台人才[2017]5655)
贵州省高层次创新型人才项目(No.黔人领发[2020]4号)
贵州省中医药管理局中医药、民族医药科学技术研究课题(No.QZYY-2020-083)
贵州中医药大学博士启动基金项目(No.贵中医博士基金[2017]1号)。
关键词
了哥王
苗药
肾毒性
乙醇总提取物
乙酸乙酯部位
石油醚部位
大鼠
Wikstroemia indica
Miao medicine
nephrotoxicity
total ethanol extract
ethyl acetate fraction
petroleum ether fraction
rat