天山堇菜总黄酮抗炎和免疫调节作用研究  

Study on the Anti-inflammatory and Immunoregulatory Effects of Total Flavonoids from Viola Tianshanica Maxim

作　　者：王雪 司丽君 刘燕 黄华 Wang Xue;Si Lijun;Liu Yan;Huang Hua(Xinjiang Institute of Materia Medica,Urumqi 830004)

机构地区：[1]新疆维吾尔自治区药物研究所,乌鲁木齐830004

出　　处：《中药药理与临床》2021年第2期49-54,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：新疆维吾尔自治区自然科学基金面上项目(编号:2017D01A71)。

摘　　要：目的:考察天山堇菜总黄酮提取物抗炎和免疫调节作用。方法:采用脂多糖(LPS)诱导小鼠腹腔巨噬细胞RAW264.7炎症模型,Griess法检测NO分泌水平,DCFH探针法检测活性氧(ROS)的含量,ELISA法检测细胞因子TNF-α、IL-6、IL-1β的含量,以及采用酶学实验检测天山堇菜总黄酮对环氧酶(COX)以及5-脂氧酶(5-LO)的抑制活性来考察天山堇菜总黄酮的抗炎作用;通过MTS法及ELISA法检测天山堇菜总黄酮对静息期淋巴细胞的影响,对刀豆蛋白(ConA)、LPS分别诱导小鼠T、B淋巴细胞增殖,以及对T淋巴细胞分泌细胞因子IL-2、IL-6、IFN-γ的影响来考察天山堇菜总黄酮提取物的免疫调节作用。结果:在LPS诱导的RAW264.7炎症模型中,与空白对照组比较,LPS模型对照组NO、TNF-α、IL-6、IL-1β含量以及活性氧ROS水平显著升高(P<0.01),与LPS模型对照组比较,天山堇菜总黄酮在0.78μg/mL~12.5μg/mL均能明显降低LPS致RAW 264.7分泌炎症介质NO和促炎症因子TNF-α、IL-6、IL-1β的水平(P<0.05或P<0.01),在3.12μg/mL~12.5μg/mL可显著降低活性氧ROS水平(P<0.01);天山堇菜总黄酮对COX-1、COX-2具有显著的抑制活性,IC_(50)分别为:13.5μg/mL, 0.73μg/mL,对5-LO也具有一定的抑制活性,其IC_(50)为334.3μg/mL;此外,天山堇菜总黄酮在6.25μg/mL~50μg/mL能显著促进静息期脾淋巴细胞增殖(P<0.01)。在T、B淋巴细胞增殖实验中,与空白对照组比较,ConA模型对照组、LPS模型对照组分别能显著促进小鼠T、B淋巴细胞增殖(P<0.01),与模型对照组比较,天山堇菜总黄酮在6.25μg/mL~200μg/mL对ConA、LPS分别诱导的小鼠T、B淋巴细胞增殖具有明显拮抗作用(P<0.05或P<0.01),在12.5μg/mL~200μg/mL明显抑制T淋巴细胞分泌IL-2、IL-6、IFN-γ(P<0.05或P<0.01)。结论:天山堇菜总黄酮具有较好的抗炎活性并且具有双向免疫调节作用,其机制有待进一步研究。Objective: To investigate the anti-inflammatory and the immunoregulatory effects of total flavonoids(TSMF) from Viola tianshanica Maxim. Methods: The inflammatory model of RAW 264.7 macrophage cells was established by LPS. The concent of NO was detected by Griess method, the intracellular reactive oxidative stress(ROS) was detected by using DCFH-probe method, the contents of TNF-α, IL-6 and IL-1β were determined by ELISA, the inhibitory activity of TSMF against cyclooxygenase(COX) and 5-Lipoxygenase were detected by enzymatic assay, in order to study its anti-inflammatory effect. Furthermore, MTS and ELISA method were used to determine the effects of TSMF on resting lymphocytes, the effect on proliferations of T and B lymphocytes induced by ConA and LPS, respectively, and the secretions of IL-2, IL-6 and IFN-γ in T lymphocytes.Results: Compared with the control group, the levels of NO, ROS,TNF-α, IL-6 and IL-1β were increased in RAW 264.7 inflammatory cell model group,(P<0.01). Compared with the model group, at the doses of 0.78 μg/mL~12.5 μg/mL, TSMF significantly inhibited the secretions of NO, TNF-α, IL-6 and IL-1β(P<0.05 or P<0.01), at the doses of 3.125 μg/mL~12.5 μg/mL, TSMF obviously reduced the content of ROS(P<0.01). In addition, TSMF showed the significantly inhibitory activities on cox-1 and cox-2, the IC_(50) s were 13.5 μg/mL and 0.73 μg/mL, respectively. TSMF also inhibited the activity of 5-LO, the IC_(50) was 334.3 μg/mL. At the doses of 6.25 μg/mL~50 μg/mL, TSMF significantly promoted the resting lymphocyte proliferation(P<0.01). In T and B lymphocyte proliferation test, compared with the control group, T and B lymphocyte proliferations were stimulated in ConA or LPS model groups(P<0.01). Compared with the model group, TSMF at the doses of 6.25 μg/mL~50 μg/mL significantly inhibited T and B lymphocyte proliferations(P<0.05 or P<0.01).TSMF at the doses of 12.5 μg/mL~200 μg/mL obviously inhibited the secretions of IL-2, IL-6 and IFN-γ(P<0.05 or P<0.01). Conclusion: TSMF has goo

关 键 词：天山堇菜 总黄酮 抗炎 免疫调节 

分 类 号：R285.5[医药卫生—中药学]