摘要
为探讨槲皮苷(quercitrin, QI)联合电针(electroacupuncture, EA)刺激对免疫抑制小鼠免疫功能的影响,将实验小鼠随机分为对照组(Control)、环磷酰胺(cyclophosphamide,CTX)模型组、QI组和QI+EA联合治疗组。用CTX建立免疫抑制小鼠模型后,第2 d起各组分别给予生理盐水、QI灌胃及QI灌胃联合EA刺激,连续治疗14 d。治疗结束后,称重法测定胸腺及脾脏指数;HE染色法观察脾脏组织学变化,吉姆萨染色法测定腹腔巨噬细胞吞噬百分率及吞噬指数;ELISA检测小鼠血浆IgG和IgM、IL-2和IL-4水平以及抗氧化相关指标如超氧化物歧化酶(superoxide diamutase, SOD)、丙二醛(malonaldehyde, MDA)、过氧化氢酶(catalase, CAT)水平;FACS检测脾脏CD4^+、CD8^+T细胞亚群以及组织活性氧类(reactive oxygen species, ROS)水平;Western blotting检测脾脏组织Nrf2总蛋白及核蛋白、总NQO1、HO-1蛋白、NF-κB p65及TLR2蛋白的表达水平。结果显示,CTX组的免疫器官指数、巨噬细胞吞噬百分率及吞噬指数、IgG、IgM、IL-2、IL-4、SOD、CAT及CD4^+、CD8^+T细胞亚群等指标均较对照组明显降低(P<0.05),而脾脏组织损伤程度、MDA、ROS及Nrf2、NQO1、HO-1、NF-κB p65、TLR2水平则均显著升高(P<0.000 1);相较于CTX组,QI组小鼠免疫功能相关指标及SOD、CAT水平和Nrf2、NQO1、HO-1蛋白水平均显著升高(P<0.05),而脾脏组织损伤程度、MDA、ROS及NF-κB p65、TLR2蛋白水平均显著下降(P<0.05);联合EA可进一步加强QI的上述作用(P<0.000 1)。由此,EA联合QI对免疫抑制小鼠有保护作用,可通过提高免疫器官功能及抗氧化水平促进小鼠的免疫力,具体作用机制可能是通过激活抗氧化应激Nrf2/ARE通路而起到提高免疫力的作用。
To investigate the effect of quercitrin combined with electroacupuncture on immune function of immunosuppressed mice, mice were randomly divided into control group, cyclophosphamide model group, quercitrin group and quercitrin+electroacupuncture group. Immunosuppressive mice model were established by CTX, and from the next day, each group was given saline, quercitrin by gavage and quercitrin by gavage combined with electroacupuncture stimulation respectively for 14 days. After the treatment, indexes of thymus and spleen were measured by weighing method;spleen tissue was observed by HE staining;macrophage phagocytosis percentage and phagocytosis index were measured by Giemsa staining;ELISA was used to test mice serum IgG, IgM, IL-2, IL-4, SOD, MDA, CAT;FACS was used to test spleen CD4^+, CD8^+T cell percentages and ROS levels;Western blotting was used to quantify spleen Nrf2, NQO1, HO-1, NF-κB p65 and TLR2. The results showed that the indexes of the thymus and the spleen, macrophage phagocytosis percentage and phagocytosis index, IgG, IgM, IL-2, IL-4, SOD, CAT, CD4^+, CD8^+T lymphocytes in CTX group were significantly lower than those in control group(P < 0.05), while the levels of immune organ tissue injury, MDA, ROS and Nrf2, NQO1, HO-1, NF-κB p65, TLR2 were significantly higher than those in control group(P < 0.000 1). Compared with CTX group, immune function related indexes, the levels of SOD, CAT, Nrf2, NQO1 and HO-1 protein in quercitrin group were significantly higher(P < 0.05), while the degree of spleen tissue injury, the levels of MDA, ROS, Nrf2, NQO1, HO-1, NF-κB p65 and TLR2 protein were significantly lower(P < 0.05). The above effects of quercitrin could be further enhanced by combined electroacupuncture therapy(P < 0.0001). Therefore, electroacupuncture combined with quercitrin has immunoprotective effect on immunosuppressed mice. It can improve the immune function of the immunosuppressive mice by improving the function of immune organs and enhancement of antioxidant levels. The possible mechanism
作者
陈龙云
周艳艳
陈会敏
徐安莉
徐建民
赵敏
CHEN Yun-long;ZHOU Yan-yan;CHEN Hui-min;XU An-li;XU Jian-min;ZHAO Min(Hubei University of Traditional Chinese Medicine,Wuhan 430000,China)
出处
《现代免疫学》
CAS
CSCD
北大核心
2019年第5期361-369,共9页
Current Immunology
基金
国家自然科学基金青年基金项目(31700152)
2016湖北中医药大学青苗计划(2016ZZX023)
关键词
电针
槲皮苷
环磷酰胺
免疫抑制
electroacupuncture
quercitrin
cyclophosphamide
immunosuppression
