BLys和TLR-4在系统性红斑狼疮转基因小鼠模型中的作用及可能机制

发布时间：2018-04-13 20:03

本文选题：Toll样受体- + B淋巴细胞刺激因子　；参考：《免疫学杂志》2017年09期

【摘要】：目的研究Toll样受体-4(Toll-like receptors-4,TLR-4)与B淋巴细胞刺激因子(B lymphocyte stimulator,BLys)在系统性红斑狼疮(systemic lupus erythematosus,SLE)转基因小鼠模型中的作用及可能机制,为SLE的治疗提供新靶点。方法选择SPF级EB病毒膜抗原BLLF1转基因雌性小鼠共30只,随机分成空白对照组、BLys阻断剂组和TLR-4阻断剂组各10只,选择SPF级野生型小鼠为野生对照组。BLys阻断剂组应用抗BR3单克隆抗体500 mg/d,腹腔注射连续10 d,TLR-4阻断剂组应用抗人TLR-4抗体250 mg/d,腹腔注射连续10 d,野生对照组和空白对照组腹腔注射等量生理盐水。继续喂养10 d后无菌取尾静脉血,RT-PCR法检测TLR-4 m RNA水平,ELISA法检测白细胞介素-6(interleukin-6,IL-6)、IL-17、肿瘤坏死因子-α(tumor necrosis factor alpha,TNF-α)和IL-2水平,金标免疫斑点法检测抗ds DNA抗体滴度,常规生化法检测补体C3(complement 3)、血沉(erythrocyte sedimentation rate,ESR)和C反应蛋白(C-reaction protein,CRP)水平,对小鼠肾脏进行HE和Masson染色。结果野生对照组和BLys阻断剂组,空白对照组和TLR-4阻断剂组外周血中单个核细胞(PBMC)的BLys无统计学差异(P0.05);与野生对照组和BLys阻断剂组比,空白对照组和TLR-4阻断剂组PBMC的BLys明显增高(P0.05)。提示EB病毒膜抗原BLLF1转基因小鼠为SLE模型,BLys阻断剂可阻断PBMC的BLys的表达。野生对照组和空白对照组干预前后的TLR-4 m RNA、IL-6、IL-17、TNF-α、IL-2、抗ds DNA抗体、C3、ESR和CRP表达无明显差异(P0.05);空白对照组、BLys阻断剂组和TLR-4阻断剂组干预前外周血TLR-4 m RNA、IL-6、IL-17、TNF-α、抗ds DNA抗体、C3、ESR和CRP表达无明显差异(P0.05)。干预后,BLys阻断剂组和TLR-4阻断剂组的TLR-4 m RNA、IL-6、IL-17、TNF-α、抗ds DNA抗体、C3、ESR和CRP较干预前明显降低(P0.05),IL-2较干预前明显增高(P0.05)。与野生对照组比,其余组别的TLR-4 m RNA、IL-6、IL-17、TNF-α、IL-2、C3、ESR和CRP表达干预前后均存在差异(P0.05)。与空白对照组比,BLys阻断剂和TLR-4阻断剂组HE和Masson染色均有明显改善。结论BLys和TLR-4可能是参与SLE的发生发展,机理可能与调节免疫炎症反应有关。
[Abstract]:Objective to study the role and possible mechanism of Toll like receptor -4 Toll-like receptors-4 (TLR-4) and B lymphocyte stimulating factor B lymphocyte stimulator (BLys4) in systemic lupus erythematosus (SLE) transgenic mice, and to provide a new target for the treatment of SLE.Methods Thirty SPF grade EB virus membrane antigen BLLF1 transgenic female mice were randomly divided into control group (n = 10) and TLR-4 blocker group (n = 10).The wild-type mice of SPF grade were selected as wild control group. BLys antagonist group was treated with anti- monoclonal antibody 500mg / d, intraperitoneal injection of anti-human TLR-4 antibody 250mg / d, intraperitoneal injection of anti-human TLR-4 antibody 250mg / d, wild control group and blank control group.Group A received intraperitoneal injection of the same amount of normal saline.After 10 days of continuous feeding, the levels of TLR-4 m RNA were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) in sterile caudal vein. The levels of IL-17, TNF- 伪 and IL-2 in tumor necrosis factor were detected by Elisa, and the titer of anti-DS DNA antibody was detected by immunoblot assay.Routine biochemical method was used to detect the levels of complement C3(complement 3, erythrocyte sedimentation rsr) and C-reactive protein C reaction protein (CRP) in the erythrocyte of mice. The kidneys of mice were stained with HE and Masson.Results there was no significant difference in BLys in peripheral blood mononuclear cells between wild control group and BLys blocker group, blank control group and TLR-4 blocker group. Compared with wild control group and BLys blocker group, the BLys of PBMC in blank control group and TLR-4 blocker group was significantly higher than that in control group and TLR-4 blocker group.It is suggested that EB virus membrane antigen BLLF1 transgenic mice can block the expression of PBMC BLys in SLE model.There was no significant difference in the expression of TLR-4 m RNA-IL-17TNF- 伪 and anti-DS DNA antibody C3TSR and CRP between the wild control group and the blank control group before and after intervention, but there was no significant difference in the expression of TLR-4 m RNA-6IL-17TNF- 伪 and anti-DS DNA antibody C3NSR and CRP between the blank control group and the TLR-4 blocker group before and after intervention.After intervention, the levels of TLR-4 m RNAs, IL-6 and IL-17 TNF- 伪, anti-DS DNA antibody C _ 3C _ 3N _ (2) and CRP in the TLR-4 antagonist group were significantly lower than those before the intervention, and the levels of P0.05 and IL-2 were significantly higher than those before the intervention.Compared with the wild control group, there were significant differences in the expression of TLR-4 m RNA-IL-6, IL-17, TNF- 伪, IL-2C3, ESR and CRP between the other groups before and after intervention (P 0.05).Compared with the blank control group, the HE and Masson staining of the two groups were significantly improved.Conclusion BLys and TLR-4 may be involved in the development of SLE, and the mechanism may be related to the regulation of immune inflammation.
【作者单位】：山东中医药大学附属医院检验科;
【分类号】：R-332;R593.241

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