参苓白术散对感染鼠伤寒沙门菌小鼠的免疫调节作用
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摘要
目的探讨参苓白术散对感染鼠伤寒沙门菌小鼠的抑菌作用及免疫调节机制。方法用鼠伤寒沙门菌标准菌株(STM,50115)制备STM小鼠模型,造模后给予小鼠参苓白术散(1 g/ml)或庆大霉素(2 mg/ml)连续灌胃10 d,观察小鼠盲肠病理改变,检测盲肠中TNF-α、IL-1β、IL-6、TLR4、MyD88、NF-κB mRNA水平及TLR4、MyD88、NF-κB蛋白含量,测定淋巴结中CD4~+T、CD8~+T及CD4~+IFN-γ和CD8~+IFN-γ细胞比例。结果与对照组比较,STM模型组TNF-α、IL-1β、IL-6及TLR4、MyD88、NF-κB mRNA水平升高(P<0.01),参苓白术散组和庆大霉素组较比STM模型组呈下降趋势(P<0.05);STM模型组TLR4、MyD88、NF-κB蛋白含量高于对照组(P<0.01),参苓白术散组和庆大霉素组低于STM模型组(P<0.05);STM模型组CD4~+T、CD8~+T及CD4~+IFN-γ和CD8~+IFN-γ细胞比例均高于对照组(P<0.05),参苓白术散组高于STM模型组(P<0.05),庆大霉素组较STM模型组有下降趋势(P>0.05)。结论参苓白术散对STM感染具有一定治疗效果,治疗机制可能与下调TLR4-MyD88-NF-κB信号诱导的炎性细胞因子分泌及上调CD4~+T、CD8~+T比例及功能有关。
Objective To investigate the antibacterial effect and immunoregulatory mechanism of Shenqi Baizhu San on mice infected with Salmonella typhimurium(STM). Methods The models of Salmonella typhimurium were constructed using standard strains(STM, 50115). After the models were established, the mice of the cecal were observed. The m RNA expression of inflammatory cytokines of TNF-α, IL-1β, IL-6, TLR4, MyD88, and NF-κB were detected. Then we analyzed the protein expression of TLR4, My D88, and NF-κB, and the proportion of CD4~+T, CD8~+T, CD4~+IFN-γ and CD8~+IFN-γ cells in the lymph nodes. Results Compared with control group, the levels of TNF-α, IL-1β, IL-6, TLR4, MyD88, and NF-κB mRNA were higher in STM model group(P <0.01), while those cytokines in Shenlin Baizhu San group and gentamicin group were lower than in STM model group(P<0.05). The protein concentration of TLR4, MyD88 and NF-κB in STM model group were higher than that in control group(P<0.01), and lower in Shenlin Baizhu San group and gentamicin group than in STM model group(P<0.05). The proportions of CD4~+T, CD8~+T, CD4~+IFN-γ and CD8~+IFN-γ cells in STM model group were higher than those in control group(P<0.05), and those cells in Shenlin Baizhu San group were higher than that in STM model group(P<0.05). Compared with STM model group, the proportions of CD4~+T, CD8~+T, CD4~+IFN-γ and CD8~+IFN-γ cells in gentamicin group had a downward trend(P >0.05). Conclusion Shenqi Baizhu San has a therapeutic effect on STM infection, and its therapeutic mechanism may be related to the down-regulation of the secretion of various inflammatory cytokines induced by TLR4-MyD88-NF-κB signaling pathway, and the up-regulation of percentage and function of CD4~+T and CD8~+T cells.
Objective To investigate the antibacterial effect and immunoregulatory mechanism of Shenqi Baizhu San on mice infected with Salmonella typhimurium(STM). Methods The models of Salmonella typhimurium were constructed using standard strains(STM, 50115). After the models were established, the mice of the cecal were observed. The m RNA expression of inflammatory cytokines of TNF-α, IL-1β, IL-6, TLR4, MyD88, and NF-κB were detected. Then we analyzed the protein expression of TLR4, My D88, and NF-κB, and the proportion of CD4~+T, CD8~+T, CD4~+IFN-γ and CD8~+IFN-γ cells in the lymph nodes. Results Compared with control group, the levels of TNF-α, IL-1β, IL-6, TLR4, MyD88, and NF-κB mRNA were higher in STM model group(P <0.01), while those cytokines in Shenlin Baizhu San group and gentamicin group were lower than in STM model group(P<0.05). The protein concentration of TLR4, MyD88 and NF-κB in STM model group were higher than that in control group(P<0.01), and lower in Shenlin Baizhu San group and gentamicin group than in STM model group(P<0.05). The proportions of CD4~+T, CD8~+T, CD4~+IFN-γ and CD8~+IFN-γ cells in STM model group were higher than those in control group(P<0.05), and those cells in Shenlin Baizhu San group were higher than that in STM model group(P<0.05). Compared with STM model group, the proportions of CD4~+T, CD8~+T, CD4~+IFN-γ and CD8~+IFN-γ cells in gentamicin group had a downward trend(P >0.05). Conclusion Shenqi Baizhu San has a therapeutic effect on STM infection, and its therapeutic mechanism may be related to the down-regulation of the secretion of various inflammatory cytokines induced by TLR4-MyD88-NF-κB signaling pathway, and the up-regulation of percentage and function of CD4~+T and CD8~+T cells.
引文
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[15]李姿慧,王键,蔡荣林,等.参苓白术散对脾虚湿困型溃疡性结肠炎大鼠结肠组织NF-κB p65蛋白表达及相关炎性因子的影响[J].北京中医药大学学报,2015,38(5):315-317,360-361.
[16]李忠.小鼠肺癌免疫细胞对参苓白术散敏感效果的研究[J].海南医学院学报,2013,19(6):738-742,745.
[2]姚营,王丽丽,徐永平,等.小鼠对鼠伤寒沙门菌感染免疫应答的研究进展[J].现代生物医学进展,2015,15(29):5773-5775,5789.
[3]Galen JE,Buskirk AD,Tennant SM,et al.Live attenuated human salmonella vaccine candidates:tracking the pathogen in natural infection and stimulation of host immunity[J].EcoSal Plus,2016,7(1):1-26.
[4]鄢鹤铭,邓锦星,陈洁,等.参苓白术散对肠道菌群及免疫调节作用的探讨[J].新中医,2015,47(11):1-2.
[5]李晓冰,崔利宏,陈玉龙,等.参苓白术散对溃疡性结肠炎小鼠肠道调节性T细胞免疫调节作用[J].中成药,2014,36(6):1295-1297.
[6]梁菊玲.参苓白术散治疗肠道菌群失调相关疾病研究进展[J].亚太传统医药2017,13(13):56-58.
[7]姚营,李晓宇,徐永平,等.鼠伤寒沙门菌诱导昆明小鼠肠道感染模型的建立[J].现代生物医学进展,2014,14(28):5401-5406.
[8]Acurcio LB,Sandes S,Bastos RW,et al.Milk fermented by Lactobacillus species from Brazilian artisanal cheese protect germ-freemice against Salmonella Typhimurium infection[J].Beneficial Microbes,2017,8(4):579-588.
[9]Wu SY,Wang LD,Xu GM,et al.Spv locus aggravates salmonella infection of zebrafish adult by inducing Th1/Th2 shift to Th2 polarization[J].Fish and Shellfish Immunology,2017,67:684-691.
[10]Johnston DG,Corr SC.Toll-Like receptor signalling and the control of intestinal barrier function[J].Methods in Molecular Biology(Clifton,N.J.),2016,1390:287-300.
[11]宋惠凤,金火星.参苓白术散的现代临床应用概述[J].中成药,2013,35(2):379-383.
[12]韦德锋,韦维,刘熙荣,等.参苓白术散治疗脾虚型溃疡性结肠炎作用机制与临床研究概述[J].辽宁中医药大学学报,2018,20(9):217-220.
[13]战仁慧,张建.Toll样受体在伤寒沙门氏菌感染过程中的作用机制[J].中国免疫学杂志,2013,29(10):1098-1102.
[14]Batista J,Ralph MT,Vaz RV,et al.Plant lectins Con Br and CFLmodulate expression toll-like receptors,pro-inflammatory cytokines and reduce the bacterial burden in macrophages infected with Salmonella enterica serovar Typhimurium[J].Phytomedicine,2017,25:52-60.
[15]李姿慧,王键,蔡荣林,等.参苓白术散对脾虚湿困型溃疡性结肠炎大鼠结肠组织NF-κB p65蛋白表达及相关炎性因子的影响[J].北京中医药大学学报,2015,38(5):315-317,360-361.
[16]李忠.小鼠肺癌免疫细胞对参苓白术散敏感效果的研究[J].海南医学院学报,2013,19(6):738-742,745.