中药复方多糖对不同MHC B-LβⅡ基因型鸡淋巴细胞中TLR4介导的MyD88依赖性途径的影响

doi:10.16431/j.cnki.1671-7236.2018.03.025

摘要/Abstract

摘要：

试验旨在探讨不同剂量中药复方多糖在不同MHC B-Lβ Ⅱ基因型鸡淋巴细胞中对TLR4及下游MyD88依赖性信号转导通路主要元件的影响。采用PCR-SSCP方法将500羽京红1号蛋鸡按MHC B-Lβ Ⅱ基因型分组，采集不同MHC B-Lβ Ⅱ基因型鸡外周血淋巴细胞，分别加入终浓度为100、75、50、0 μg/mL（高、中、低剂量组和对照组）中药复方多糖共培养16、24、32、48 h收集细胞，应用实时荧光定量PCR方法检测不同MHC B-Lβ Ⅱ基因型鸡TLR4、MyD88、TNAF-6基因的表达。结果显示，与对照组相比，中药复方多糖能显著增强不同MHC B-Lβ Ⅱ基因型鸡TLR4、MyD88、TNAF-6 mRNA的表达量（P < 0.05）；中、低剂量组AA基因型鸡TLR4、MyD88、TNAF-6基因的表达量高于高剂量组（培养16 h时TLR4基因除外），高剂量组BB基因型TLR4、MyD88、TNAF-6基因的表达量高于其他剂量组（培养32和48 h时TLR4基因除外）），低剂量组BC基因型TLR4、MyD88、TNAF-6基因的表达量高于其他剂量组（培养16 h时TLR4基因除外）。结果表明，中药复方多糖发挥机体免疫调节作用的机制是通过与淋巴细胞表面TLR4结合，激活下游MyD88依赖性信号转导通路，调控细胞免疫，且不同MH-C B-Lβ Ⅱ基因型鸡淋巴细胞最适中药复方多糖免疫调节剂量不同。

关键词: 中药复方多糖; MHC B-LβⅡ基因; 鸡淋巴细胞; TLR4; MyD88依赖性信号转导通路

Abstract:

To investigate the effects of different doses of compound Chinese herbal medicinal polysaccharides (cCHMPS) on TLR4 and downstream MyD88 dependent signal transduction pathway components in chicken lymphocytes of different MHC B-LβⅡ genotypes,PCR-SSCP technique was applied to group layer according to different MHC B-LβⅡ genotypes.The peripheral blood lymphocytes of chicken with different MHC B-LβⅡ genotypes were collected,and added with 100,75,50 and 0 μg/mL cCHMPS (high,middle and low dose groups and control group),respectively,then co-culturing for 16,24,32 and 48 h.The expression of TLR4,MYD88 and TRAF-6 mRNA were detected using Real-time PCR method.The results showed that compared with control group,cCHMPS could significantly improve the expression levels of TLR4,MYD88 and TRAF-6 mRNA of different MHC B-Lβ Ⅱ genotypes chickens (P < 0.05);The expression levels of TLR4,MYD88 and TRAF-6 mRNA of AA genotype chicken lymphocyte in middle and low dose groups were higher than those of high dose group (except TLR4 gene cultured for 16 h);The expression of TLR4,MyD88 and TNAF-6 mRNA of BB genotype in high dose group were higher than those of other dose groups (except TLR4 gene cultured for 32 and 48 h);The expression of TLR4,MyD88 and TNAF-6 mRNA of BC genotype in low dose were higher than that of other dose groups (except TLR4 gene cultured for 16 h).There results indicated that cCHMPS played an important role in the body’s immune regulatory mechanism by binding to TLR4 in the surface of lymphocytes,activating the downstream MyD88-dependent signal transduction pathway,regulating cellular immunity,and cCHMPS optimum immunomodulatory does were different in each MHC B-Lβ Ⅱ genotype chickens.

Key words: compound Chinese herbal medicinal polysaccharides (cCHMPS); MHC B-Lβ Ⅱ gene; chicken lymphocyte; TLR4; MyD88 dependent signaling pathway

中图分类号:

S853.7

马昭, 刘钢, 杨莉, 刘晓婷, 乔彦杰, 朱晓庆, 谷新利. 中药复方多糖对不同MHC B-LβⅡ基因型鸡淋巴细胞中TLR4介导的MyD88依赖性途径的影响[J]. 《中国畜牧兽医》, 2018, 45(3): 754-762.

MA Zhao, LIU Gang, YANG Li, LIU Xiaoting, QIAO Yanjie, ZHU Xiaoqing, GU Xinli. Effect of Compound Chinese Herbal Medicinal Polysaccharides on MyD88 Dependent Pathway Mediated by TLR4 in Lymphocytes of Chicken with Different MHC B-LβⅡ Genotypes[J]. , 2018, 45(3): 754-762.

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