大黄附子汤对BALB/c小鼠腹腔巨噬细胞功能的影响

Effects of Dahuang Fuzi Decoction on the Functions of Peritoneal Macrophages in BALB/c Mice

目的:研究大黄附子汤对BALB/c小鼠腹腔巨噬细胞功能的影响。方法:制备小鼠腹腔巨噬细胞,纯化后以5×105/mL细胞接种于96孔培养板或6孔板中,分为:细胞对照组、脂多糖(lipopolysaccharide,LPS)模型组(10 mg·L-1)、大黄附子汤组(终质量浓度为25,50,100,200,400,800 mg·L-1),在细胞贴壁过夜后,正常组给予不含血清的培养液,其余各组给予LPS(10 mg·L-1),大黄附子汤组同时给予不同浓度药物,48 h后测细胞存活率,ELISA法测肿瘤坏死因子-α(TNF-α),白介素-6(IL-6)水平,黄嘌呤氧化酶法(羟氨法)测细胞上清液中超氧化物歧化酶(SOD)活力,硫代巴比妥酸法测丙二醛(MDA)含量,Griess法测一氧化氮(NO)水平。结果:在25～400 mg·L-1内,大黄附子汤对正常细胞代谢MTT活力无显著影响;剂量达50 mg·L-1即能抑制LPS诱导NO的生成(P<0.01),100 mg·L-1时能降低MDA含量(P<0.01),增强SOD酶活力(P<0.05),并能显著抑制LPS诱导TNF-α,IL-6等细胞因子的合成(P<0.01)。结论:大黄附子汤能有效调节BALB/c小鼠腹腔巨噬细胞免疫及抗氧化功能,改善LPS对BALB/c小鼠腹腔巨噬细胞的诱导作用。

Objective： To investigate the effects of Dahuang Fuzi decoction on the function of peritoneal macrophages inmice.Method： Macrophages were seeded onto glass cover slips（5×105 cells/mL）.Different concentrations of Dahuang Fuzi decoction（25,50,100,200,400,800 mg·L-1） were added to the normal or LPS-activited peritoneal macrophages.Metabolic activity of peritoneal macrophages was measured by MTT assay,nitric oxide was detected by Griess reagent,malondiadehyde（MDA） content was detected by hydroxylamine method,superoxide dismutase（SOD） activities was assayed by thiobarbituric acid method,concentrations of tumor necrosis factor-α（TNF-α） and interleukin-6（IL-6） in supernatants were determined by using quantitative sandwich enzymelinked immunosorbent assay（ELISA）kits.Result： Dahuang Fuzi decoction had no obvious effect on the metabolic activity of peritoneal macrophages,but could markedly inhibit the secretion of NO,MDA,TNF-α and IL-6（P0.01）,furthermore,Dahuang Fuzi decoction could improved the production of SOD（P0.05）.Conclusion： Dahuang Fuzi decoction can promote the immune and antioxidant capacity of peritoneal macrophagesin BALB/c mice.