基于巨噬细胞吞噬及分泌功能的清金化痰汤生物限值测定  

Biological Evaluation for Quality Control of Water Extract of Qingjin Huatantang Based on Phagocytic and Secretory Functions of Macrophages

作　　者：张琼玲 孙正霄 肖顺丽 丁世兰 许浚 张铁军 游云[1] ZHANG Qiong-ling;SUN Zheng-xiao;XIAO Shun-li;DING Shi-lan;XU Jun;ZHANG Tie-jun;YOU Yun(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Tianjin Institute of Pharmaceutical Research,Tianjin 300301,China)

机构地区：[1]中国中医科学院中药研究所,北京100700 [2]天津药物研究院有限公司,天津300301

出　　处：《中国实验方剂学杂志》2021年第24期10-16,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家“重大新药创制”科技重大专项(2018ZX09721004-006-003)。

摘　　要：目的:建立基于小鼠巨噬细胞RAW264.7吞噬及分泌功能的清金化痰汤水提物冻干粉生物限值测定方法,从生物活性层面对清金化痰汤进行质量控制。方法:建立巨噬细胞RAW264.7吞噬模型和炎症模型,以清金化痰汤水提物冻干粉对巨噬细胞RAW264.7吞噬指数和白细胞介素-6(IL-6)分泌量的抑制率为指标,考察清金化痰汤水提物冻干粉的生物活性,并寻找生物限值。结果:巨噬细胞优选接种密度为3×10^(5)个/mL,药物作用时间24 h,脂多糖(LPS)质量浓度1 mg·L^(-1);当清金化痰汤水提物冻干粉质量浓度为500 mg·L^(-1)时,对RAW264.7巨噬细胞既无毒性也无明显促增殖作用,在此浓度下,对RAW264.7巨噬细胞吞噬中性红有明显促进作用,吞噬指数均>113%,且对LPS诱导RAW264.7巨噬细胞分泌IL-6呈现出显著且稳定的抑制作用,抑制率>45%。结论:结合清金化痰汤抗炎及免疫调节的药理活性,建立了基于RAW264.7巨噬细胞吞噬及分泌功能的生物限值评价方法,在设定的实验条件下,确定500 mg·L^(-1)为限值浓度,在该限值浓度下,清金化痰汤水提物对巨噬细胞的吞噬指数有明显的促进作用或对LPS诱导的RAW264.7巨噬细胞IL-6的分泌量有明显抑制作用,均可判断为质量合格。Objective:To establish a method for evaluating the biological activity of water extract lyophilized powder of Qingjin Huatantang based on the phagocytic and secretory functions of macrophages,and to control the quality of this formula from the biological activity level.Method:The phagocytic and inflammation models of RAW264.7 macrophages were established,the inhibition rates of water extract lyophilized powder of Qingjin Huatantang on interleukin-6(IL-6)secretion and phagocytic index of neutral red of RAW264.7 macrophages were chosen as indicators to investigate the biological activity of Qingjin Huatantang,and the biological limit was searched.Result:The optimal inoculation density of RAW264.7 macrophages was 3×10^(5) pcs/mL,and the concentration of lipopolysaccharide(LPS) was 1 mg·L^(-1) after treatment for 24 h.When the concentration was 500 mg·L^(-1),water extract lyophilized powder of Qingjin Huatantang had no toxicity and no obvious promotion effect on the proliferation of RAW264.7 macrophages,and at this concentration,the phagocytosis of RAW264.7 macrophages for neutral red was significantly promoted,the phagocytic index was >113%.In addition,the lyophilized powder had a significant and stable inhibitory effect on IL-6 secretion of RAW264.7 macrophages induced by LPS,the inhibitory rate was >45%.Conclusion:Combined with the anti-inflammatory and immunomodulatory effects of Qingjin Huatantang,this study establishes an in vitro biological limit method for evaluating the quality of water extract of Qingjin Huatantang based on the phagocytic and secretory functions of RAW264.7 macrophages,and 500 mg·L^(-1) was confirmed as the limit concentration.Under the limit concentration,Qingjin Huatantang water extract can significantly promote the phagocytic index of macrophages or significantly inhibit the secretion of IL-6 of RAW264.7 macrophages induced by LPS,which can be judged as qualified.

关 键 词：清金化痰汤 水提物 冻干粉 RAW264.7巨噬细胞 吞噬指数 白细胞介素 生物活性评价 

分 类 号：R22[医药卫生—中医基础理论] R28[医药卫生—中医学]