显齿蛇葡萄醇提物含药血清对脂多糖诱导巨噬细胞RAW264.7炎症模型的抗炎作用  

Study on Anti-Inflammatory Effects of Serum Containing Ampelopsis Grossedentata Alcohol Extract on LPS Induced RAW264.7 Inflammation Model

作　　者：邱盼子 谢谦 黄涛 欧阳波[1,2] 肖作奇 QIU Panzi;XIE Qian;HUANG Tao;OUYANG Bo;XIAO Zuoqi(Hunan Provincial Maternal and Child Health Care Hospital,Changsha Hunan 410008,China;Hunan Engineering Research Center of Traditional Chinese Medicine Clinical Formula Transformation for Women and Children,Changsha Hunan 410008,China)

机构地区：[1]湖南省妇幼保健院,湖南长沙410008 [2]妇儿中医临床验方转化湖南省工程研究中心,湖南长沙410008

出　　处：《中医药导报》2024年第10期27-31,共5页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：湖南省自然科学基金项目(2021JJ80026,2021JJ80027);长沙市科技创新平台项目(kh2005013);湖南省中医药管理局一般项目(D202044)。

摘　　要：目的:采用血清药物化学的研究方法,分析显齿蛇葡萄醇提物(AGEE)大鼠口服后的入血成分,基于入血成分研究AGEE含药血清对脂多糖(LPS)诱导RAW264.7细胞炎症的抗炎作用。方法:12只SD大鼠随机分为AGEE组和空白组,分别给予AGEE(5.4 g/kg)和纯净水灌胃,以制备含药血清和空白血清,采用HPLC-DAD分析其含药血清色谱图,通过与空白血清色谱图对比分析,探讨AGEE的体内移行成分。以RAW264.7细胞为研究对象,将细胞分为5%或15%空白血清组,5%或15%空白血清+LPS组,以及5%或15%含药血清+LPS组,按相应条件培养24 h后,采用细胞增殖-毒性检测试剂盒(CCK-8)法检测每组细胞存活率;采用酶联免疫吸附试验(ELISA)法检测各组细胞中TNF-α、IL-6、IL-10水平。结果:AGEE含药血清与空白血清比较,发现16个色谱峰,其中9种为原型成分入血,7种可能是AGEE在大鼠体内产生的代谢物。CCK-8检测结果表明,各组细胞按上述条件培养24 h后均未发现毒性。5%或15%空白血清+LPS组细胞中TNF-α、IL-6、IL-10水平高于5%或15%空白血清组(P<0.01或P<0.05)。15%含药血清+LPS组细胞中TNF-α水平低于5%空白血清+LPS组(P<0.05);5%或15%含药血清+LPS组细胞IL-6水平低于5%或15%空白血清+LPS组(P<0.01);5%或15%含药血清+LPS组细胞IL-10水平高于5%或15%空白血清+LPS组(P<0.01或P<0.05)。结论:建立了AGEE含药血清HPLC-DAD分析方法,明确了其活性物质基础。AGEE含药血清可降低LPS诱导RAW264.7细胞炎症反应。Objective:To analyze the blood components of Ampelopsis Grossedentata Alcohol Extract(AGEE)after oral administration by serum pharmacochemical method,and to study the anti-inflammatory effect of serum containing AGEE on LPS induced inflammation in RAW264.7 cells based on the blood components.Methods:Totally 12 SD rats were randomely divided into AGEE group and blank group,respectively orally given AGEE(5.4 g/kg)and distilled water to prepare drug-containing serum and blank serum.HPLC-DAD was used to analyze the chromatogram of drug-containing serum,and the internal migration components of AGEE were explored by comparing with the chromatogram of blank serum.RAW264.7 cells were divided into 5%or 15%blank serum group,5%or 15%blank serum+LPS group,and 5%or 15%serum containing AGEE+LPS group.After 24 hours of culture according to the corresponding conditions,the cell viability of each group was detected by CCK-8 method,and the levels of TNF-α,IL-6,and IL-10 in each group of cells were detected by ELISA method.Results:Compared with the blank serum,16 chromatographic peaks were found in the serum containing AGEE,among which 9 components were original components entering the blood and 7 components were possible metabolites produced by AGEE in rats.CCK-8 assay showed that there was no toxicity found in each group of cells cultured under the above conditions for 24 hours.The 5%or 15%blank serum+LPS group showed higher levels of TNF-α,IL-6 and IL-10 than 5%or 15%blank serum group(P<0.01 or P<0.05).The 15%serum containing AGEE+LPS group showed lower level of TNF-αthan 5%blank serum+LPS group(P<0.05).The 5%or 15%serum containing AGEE+LPS group showed lower levels of IL-6 than 5%or 15%blank serum+LPS group(P<0.01).The 5%or 15%containing AGEE+LPS group showed higher level of IL-10 than 5%or 15%blank serum+LPS group(P<0.01 or P<0.05).Conclusion:A method for analyzing AGEE-containing serum by HPLC-DAD was established,and the active substance basis of AGEE has been preliminarily clarified.AGEE-containing serum can significantly

关 键 词：显齿蛇葡萄 血清药物化学 RAW264.7细胞 炎症因子 巨噬细胞 

分 类 号：R285.5[医药卫生—中药学]