芪斛楂颗粒不同部位对小鼠巨噬细胞增殖和吞噬活性及细胞周期的影响  

Effects of different fractions of qihuzha granule on the proliferation,phagocytic activity,and cell cycle of mouse macrophages

作　　者：黄甫静 张金娟 冯敏 廖尚高 何迅 HUANG Fujing;ZHANG Jinjuan;FENG Min;LIAO Shanggao;HE Xun(School of Pharmacy,Guizhou Medical University,Guiyang 550025,Guizhou,China;School of Basic Medical,Guizhou Medical University,Guiyang 550025,Guizhou,China;Guizhou Hongqi Co.,Ltd.,Sunflower Pharmaceutical Group,Liupanshui 553400,Guizhou,China;School of Medicine and Health Management,Guizhou Medical University,Guiyang 550025,Guizhou,China)

机构地区：[1]贵州医科大学药学院,贵州贵阳550025 [2]贵州医科大学 [3]贵州医科大学基础医学院,贵州贵阳550025 [4]葵花药业集团贵州宏奇有限公司,贵州六盘水553400 [5]贵州医科大学医药卫生管理学院,贵州贵阳550025

出　　处：《贵州医科大学学报》2021年第11期1264-1270,共7页Journal of Guizhou Medical University

基　　金：国家自然科学基金(81360635);贵州省科技计划项目[黔科合平台人才(2020)5006]。

摘　　要：目的探讨芪斛楂颗粒不同部位对小鼠巨噬细胞增殖、吞噬活性及细胞周期的影响。方法取芪斛楂颗粒浸膏制备多糖部位(Fr.A)、非多糖水洗脱部位(Fr.B)、非多糖75%乙醇洗脱部位(Fr.C)及非多糖95%乙醇洗脱部位(Fr.D),取对数生长期的小鼠单核巨噬细胞株RAW264.7,分别加7.81、15.73、31.25、62.50、125.00、250.00、500.00及1000.00 mg/L芪斛楂颗粒Fr.A、Fr.B、Fr.C及Fr.D处理24 h,同时设空白组(不含药完全培养基);采用四唑盐(MTT)法检测各组RAW264.7细胞的吸光度(OD)值并计算细胞增殖率;取对数生长期RAW264.7细胞,分为空白组(完全培养基)、脂多糖(LPS)组(5.00 mg/L LPS)及低(30.00 mg/L)、中(60.00 mg/L)、高剂量(120.00 mg/L)Fr.A组,采用中性红吞噬试验测定各组RAW264.7细胞的OD值并计算中性红吞噬率,采用流式细胞术测定各组细胞周期。结果MTT法检测显示,与空白组相比,15.73~1000.00 mg/L Fr.A和Fr.C组、500.00~1000.00 mg/L Fr.B组RAW264.7细胞的增殖率均上升(P<0.05或P<0.01),其中Fr.A组的细胞增殖率较空白组升高最为明显,7.81~1000.00 mg/L Fr.D组细胞增殖率无变化(P>0.05);中性红吞噬试验结果显示,与空白组比较,各质量浓度Fr.A组中性红吞噬率均上升(P<0.05或P<0.01);流式细胞仪检测结果显示,与空白组比较,中、高剂量Fr.A组RAW264.7细胞G0/G1期的比例上升(P<0.05或P<0.01),S期的细胞比例无变化(P>0.05),G2/M期的细胞比例降低(P<0.05或P<0.01)。结论芪斛楂颗粒Fr.A、Fr.B及Fr.C部位能促进小鼠巨噬细胞RAW264.7的增殖,以Fr.A部位作用最强,且可促进RAW264.7细胞的吞噬能力,增加处于G0/G1期的巨噬细胞比例。Objective To explore the effect of different fractions from Qihuzha Granules on proliferation,phagocytic activity and cell cycle of mouse macrophages.Methods Qihuzha granules was used to extract the fractions:Fr.A-polysaccharide,Fr.B-non-polysaccharide eluted by water,Fr.C-non-polysaccharide eluted by 75%ethanol,and Fr.D-non-polysaccharide eluted by 95%ethanol.RAW264.7 cells at logarithmic growth phase were treated with Fr.A,Fr.B,Fr.C,and Fr.D at the concentrations of 7.81,15.73,31.25,62.50,125.00,250.00,500.00,and 1000.00 mg/L of for 24 h,respectively.RAW264.7 cells were cultured in complete culture medium as a blank group.Methyl Thiazolyl Tetrazolium(MTT)assay was applied to detect the absorbance(OD)value of each group,and calculate the cell proliferation rate of each group.Based on the treatment,RAW264.7 cells at logarithmic growth phase were divided into control group(complete culture medium),lipopolysaccharide(5.00 mg/L LPS)group,low-Fr.A(30.00 mg/L),medium-Fr.A(60.00 mg/L),and high-Fr.A dose(120.00 mg/L)groups.Neutral red staining was used to determine the OD value of each group and calculate phagocytosis rates.Flow cytometry was used to determine the cell cycle changes in each group.Results MTT assay showed that the proliferation rates of RAW264.7 cells were increased in 15.73~1000.00 mg/L Fr.A,Fr.C,and 500.00~1000.00 mg/L Fr.B groups when compared to blank group(P<0.05 or P<0.01).The increase in cell proliferation rate was the most obvious in Fr.A group.Fr.D at 7.81~1000.00 mg/L had no effect on cell proliferation rate(P>0.05).Neutral red staining revealed that phagocytosis rate in Fr.A at different concentrations were increased when compared to control group(P<0.05 or P<0.01).Flow cytometry analysis showed that when compared to control group,the ratio of RAW264.7 cells in G0/G1 phase was increased(P<0.05 or P<0.01)in the medium-and high-dose Fr.A groups,the ratio of RAW264.7cells in S phase did not change(P>0.05),and the ratio of RAW264.7 cells in G2/M phase were decreased(P<0.05 or P<0.01).Conclusion Th

关 键 词：脂多糖类 细胞周期 芪斛楂颗粒 巨噬细胞RAW264.7 增殖活性 吞噬活性 

分 类 号：R285.5[医药卫生—中药学]