TLR/NF-κB independent signaling pathway in TNF-α suppression of diabetic MyD88-knockout mice after <i>Lycium barbarum</i> polysaccharides administration

doi:10.3785/j.issn.1008-9292.2018.02.05

J Zhejiang Univ (Med Sci)

2018, Vol. 47

Issue (1): 35-40 DOI: 10.3785/j.issn.1008-9292.2018.02.05

TLR/NF-κB independent signaling pathway in TNF-α suppression of diabetic MyD88-knockout mice after Lycium barbarum polysaccharides administration

LIU Tingting(

),WANG Lingxiao,YANG Xiaohui,YAO Zhiqing,CAI Huizhen*(

)

School of Public Health and Management, Ningxia Medical University, Yinchuan 750004, China

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Abstract

Objective: To investigate the effect of Lycium barbarum polysaccharides (LBPs) on TLR/NF-κB independent pathway and serum tumor necrosis factor (TNF-α) level in diabetic MyD88-knockout mice. Methods: Diabetes was induced by feeding high-fat/high-sugar diet and injection of low-dose streptozotocin in MyD88-knockout mice. The diabetic mice were randomly divided into model group, positive control group and LBPs group. The expressions of TRAM, TRIF, TRAF6, RIP1 and TNF-α mRNA and proteins in mouse peritoneal macrophages were detected by real-time RT-PCR and Western blotting after LBPs treatment for 3 month. Serum TNF-α was determined with ELISA kit. Results: Real time RT-PCR showed that compared with model group, the relative expressions of Tram, Trif, Traf6 and Tnf-α mRNA in macrophages of LBPs group were significantly decreased and expression of Rip1 was significantly increased (all P < 0.05). Expression of TRAM, TRIF, TRAF6, RIP1 and TNF-α proteins as well as serum TNF-α level had no significant difference between LBPs group and model group (all P > 0.05). Conclusion: LBPs may not inhibit serum TNF-α level through TLR/NF-κB independent pathway.

Key words： Myeloid differentiation factor 88/physiology Signal transduction Tumor necrosis factor-alpha/biosynthesis Polysaccharides/pharmacology Lycium barbarum/chemistry Diabetes mellitus, type 2 Disease models, animal Cells, cultured

Received: 03 December 2017 Published: 12 June 2018

CLC:

R96

Corresponding Authors: CAI Huizhen E-mail: 18695272282@163.com;xingcao_c@sina.com

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	LIU Tingting
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	YANG Xiaohui
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Cite this article:

LIU Tingting,WANG Lingxiao,YANG Xiaohui,YAO Zhiqing,CAI Huizhen. TLR/NF-κB independent signaling pathway in TNF-α suppression of diabetic MyD88-knockout mice after Lycium barbarum polysaccharides administration. J Zhejiang Univ (Med Sci), 2018, 47(1): 35-40.

URL:

http://www.zjujournals.com/med/10.3785/j.issn.1008-9292.2018.02.05 OR http://www.zjujournals.com/med/Y2018/V47/I1/35

MyD88非依赖性信号通路在枸杞多糖抑制糖尿病小鼠肿瘤坏死因子α中的作用

目的: 研究枸杞多糖对髓样分化因子88（MyD88）非依赖性信号通路的影响，探讨枸杞多糖影响TNF-α生成的机制。方法: 采用高糖高脂饲料联合链脲菌素诱导MyD88基因敲除小鼠形成2型糖尿病模型。将造模成功的小鼠随机分为模型对照组、阳性对照组和枸杞多糖组，另取8只小鼠作为健康对照组。干预三个月后，采用实时定量RT-PCR和蛋白质印迹法检测小鼠腹腔巨噬细胞中TRAM、TRIF、TRAF6、RIP1和TNF-α基因和蛋白表达，ELISA法测定小鼠血清TNF-α水平。结果: 枸杞多糖抑制了糖尿病小鼠腹腔巨噬细胞中Tram、Trif、Traf6和Tnf-α的基因表达，激活了Rip1基因（均P < 0.05），但对TRAM、TRIF、TRAF6、RIP1和TNF-α蛋白表达和血清TNF-α水平无影响（均P > 0.05）。结论: 枸杞多糖可能不能通过MyD88非依赖性信号通路抑制TNF-α的生成。

关键词： 髓样分化因子88/生理学, 信号传导, 肿瘤坏死因子α/生物合成, 多糖类/药理学, 枸杞/化学, 糖尿病, 2型, 疾病模型, 动物, 细胞, 培养的

Tab 1 Primer sequences for real-time RT-PCR

Tab 2 mRNA expressions of Trif, Tram, Traf6, Rip1 and Tnf-α in mice peritoneal macrophages

Fig 1 Electropherograms of TRIF, TRAM, TRAF6, RIP1 and TNF-α expression in each group

Tab 3 Expressions of TRIF, TRAM, TRAF6, RIP1 and TNF-α protein in mice peritoneal macrophages


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