基于TLRs/NODs受体与MAPK及NF-κB信号通路的朱砂七免疫调节机制研究  

Study on immunomodulatory mechanism of Polygonum Cillinerve based on TLRs/NODs receptor and MAPK and NF-κB signaling pathway

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作  者:曹兰秀 吕娟 赵娴 王江 马莉 CAO Lan-Xiu;LU Juan;ZHAO Xian;WANG Jiang;MA Li(Department of Prescription,Shanxi University of Traditional Chinese Medicine,Xianyang 712046,China)

机构地区:[1]陕西中医药大学基础医学院方剂教研室,咸阳712046 [2]陕西中医药大学中药系,咸阳712046

出  处:《中国免疫学杂志》2019年第4期424-428,434共6页Chinese Journal of Immunology

基  金:陕西省科技厅自然科学基础研究项目(2017JM8021);陕西省教育厅基础研究项目(2010Jk476)。

摘  要:目的:本研究探讨朱砂七是否可以增强小鼠免疫力,并明确这种免疫调节机制是否与TLRs/NODs受体与MAPK及NF-κB信号通路相关。方法:建立金黄色葡萄球菌感染小鼠模型,40只雄性C57BL/6J小鼠分为四组,即正常对照组(NS)、金葡菌肺炎模型组(MRSA)、朱砂七组(PC+MRSA)、青霉素组(阳性药,PG+MRSA)。记录各组小鼠死亡率,HE染色观察组织病变,计算活菌数量,ELISA法检测血液炎性因子TNF-α、IL-6水平。Real time PCR法检测TLRs和NODs及其下游炎性因子TNF-α、IL-6 mRNA表达,进一步检测MAPK及NF-κB的基因水平表达。Western blot检测其蛋白表达水平。结果:与MRSA组比较,朱砂七组死亡率明显降低,差异有统计学意义(P<0.05);HE染色结果显示朱砂七组小鼠肺部组织炎性病变明显减轻,活菌数量减少。Real time PCR及Western blot结果显示,与正常对照组比较,MRSA组TLR2、NOD2及其下游炎性因子IL-6、TNF-αmRNA均有明显上升(P<0.05),青霉素组、朱砂七组的上升趋势显著降低,但是仍显著高于正常对照组(均P<0.05)。MRSA模型组动物MAPK、NF-κB信号通路在转录和翻译水平的改变也较为明显,朱砂七可降低MRSA暴露小鼠的MAPK、NF-κB在mRNA及蛋白的表达水平(P<0.05)。结论:朱砂七可以减轻金黄色葡萄球菌对小鼠的感染症状,且该作用可能与其调控TLRs/NODs模式识别受体及其下游炎性因子TNF-α、IL-6相关,该作用可能通过MAPK及NF-κB信号通路起作用。Objective:To investigate whether Polygonum Cillinerve could enhance the immunity of mice,and also explored whether the mechanism of the immune regulation was related with TLRs/Nods receptor and MAPK,NF-κB pathway.Methods:A mouse model of Staphylococcus aureus infection was established,40 male C57 mice were divided into four groups:control group,model group,Polygonum Cillinerve group,penicillin group.The mortality rate of mice was recorded,and tissue HE staining was used to evaluate the tissue damage of mice in each group.The number of live bacteria was calculated either,ELISA method was used to detect the level of inflammatory factor TNF-α/IL-6 in blood.The gene expression level of pattern recognition receptor TLRs and NODs and its downstream inflammatory factor TNF-α/IL-6 in mice were detected by Real time PCR and Western blot,and the gene and protein expression level of MAPK and NF-κB was further detected.Results:Polygonum Cillinerve significantly inhibited the mortality of experimental and host cells of Staphylococcus aureus infection.The inflammatory lesions in the lung tissue of mice in Polygonum Cillinerve groups were significantly reduced and the number of living bacteria was also decreased.Real time PCR and Western blot results showed that TLR2 and NOD2 and their downstream inflammatory factors IL-6 and TNF-αwere increased significantly in MRSA model group(P<0.05).Penicillin and Polygonum Cillinerve could significantly reduced this upward trend,but it was still significantly higher than that in normal group(P<0.05).Polygonum Cillinerve could reduce MAPK,NF-κB mRNA and protein expression level in MRSA exposed animals(P<0.05).Conclusion:Polygonum Cillinerve can reduce the infection symptoms of Staphylococcus aureus in mice,and this effect may be related to its regulation of TLRs/NODs pattern recognition receptor and its downstream inflammatory factor TNF-α/IL-6,which may play important role through regulate MAPK and NF-κB signaling pathway.

关 键 词:朱砂七 免疫调节 TLRS NODs MAPK NF-κB 

分 类 号:R967[医药卫生—药理学;医药卫生—药学]

 

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