摘要
目的研究黄草乌中分离得到的12个醇胺型二萜生物碱的镇痛活性和毒性,揭示黄草乌的药效物质基础。方法将不同浓度的醇胺型生物碱分别作用于H9c2细胞和PC12细胞,采用MTT法检测细胞存活率,以观察醇胺型生物碱的细胞毒性。将醇胺型生物碱作用HAPI小胶质细胞后,观察细胞强啡肽前体基因表达的改变以初步探讨其镇痛活性。分别采用小鼠醋酸扭体法和热板法观察4个醇胺型生物碱的外周和中枢镇痛作用。结果在H9c2细胞和PC12细胞上,大多数醇胺型生物碱细胞存活率均大于90%,对细胞影响较小,少数醇胺型生物碱高浓度时显示毒性。与正常组HAPI小胶质细胞比较,浓度为30μmol/L的各给药组细胞存活率均大于90%,不显示细胞毒性;除Pengshenine B和Vilmorrianine F外,其余生物碱均能提高HAPI细胞中的Prodynorphin mRNA,其中Vilmorrianine B、Vilmorrianine D和8-O-methyl-sachaconitine能够显著增加小胶质细胞的Prodynorphin mRNA(P<0.05)。与空白对照组比较,Isotalatizidine、Vilmorrianine B、Vilmorrianine D高、中剂量组,Talatisamine高剂量组对冰醋酸所致的小鼠扭体均具有抑制作用,差异具有统计学意义(P<0.05,P<0.01);与基础痛阈值比较,Isotalatizidine、Talatisamine、Vilmorrianine B、Vilmorrianine D高、中剂量组在给药15min后均能显著延长热板所致小鼠疼痛的痛阈值(P<0.05,P<0.01)。结论黄草乌中醇胺型生物碱毒性均较低,含量较高的4种成分都具有明显的镇痛活性,是黄草乌药效物质基础的重要组成部分,其镇痛作用可能是通过刺激脊髓背角的小胶质细胞释放强啡肽而产生。
Objective The analgesic activities and toxicities of twelve amino-alcohol diterpenoid alkaloids isolated from the roots of A.vilmorinianum were studied to reveal the pharmacological substance of this medicinal material.Methods Different concentra⁃tions of amino-alcohol alkaloids were applied to H9c2 cells and PC12 cells respectively,and the cell survival rate was detected by MTT method to observe the cytotoxicity of these compounds.After treating HAPI microglia with amino-alcohol alkaloids,the expression of Prodynorphin mRNA was observed to explore their analgesic activity.The peripheral and central analgesic effects of four main amino-alcohol alkaloids were observed through acetic acid writhing method and hot plate method in mice respectively.Results The survival rate under most amino-alcohol alkaloids was more than 90%on H9c2 and PC12 cells,only a few of them showed toxicity at high concentration.Compared with the normal group,the survival rate of HAPI microglia was more than 90%in all groups with the concentration of 30μmol/L.Except Pengshenine B and Vilmorrianine F,other alkaloids could increase the ex⁃pression of Prodynorphin mRNA in HAPI microglia cells,in which Vilmorrianine B,Vilmorrianine D and 8-O-methyl-sa⁃chaconitine showed significant effect(P<0.05).Compared with the blank control group,the high and middle-monitoring groups of Isotalatizidine,Vilmorrianine B and Vilmorrianine D,the high-dose groups of Talatisamine showed significant inhibito⁃ry effects on acetic acid-induced writhing in mice(P<0.05,P<0.01).Compared with the basic pain threshold,the high and middle-monitoring groups of Isotalatizidine,Talatisamine,Vilmorrianine B and Vilmorrianine D could significantly prolong the pain threshold fifteen minutes after administration in hot plate mice model(P<0.05,P<0.01).Conclusion The amino-alco⁃hol alkaloids isolated from the A.vilmorinianum have weak toxicity and potent analgesic activity.Four components with high con⁃tent have obvious analgesic activities,which are the important part o
作者
苏东雪
庄馨瑛
段小花
马晓霞
SU Dong-xue;ZHUANG Xin-ying;DUAN Xiao-hua;MA Xiao-xia(Yunnan University of Chinese Medicine,Kunming Yunnan 650500,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2023年第12期2826-2830,共5页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(82160746)
云南省“万人计划”青年拔尖人才专项(YNWR-QNBJ-2019-011)
云南省科技计划项目(2019ZF003)。
关键词
黄草乌
醇胺型生物碱
镇痛活性
强啡肽
小胶质细胞
Aconitum vilmorinianum
Amino-alcohol alkaloid
Analgesic activity
Dynorphin
Microglial cell
