文章摘要
徐旭 史剑 谈春业 刘锦波 何小舟.慢性移植性肾病大鼠脾脏中辅助性T细胞和B 细胞免疫状态的研究*[J].,2016,16(32):6220-6225
慢性移植性肾病大鼠脾脏中辅助性T细胞和B 细胞免疫状态的研究*
The Immune Status of Th and B Cells in the Rat Spleen during ChronicAllograft Nephropathy
  
DOI:
中文关键词: Th 细胞  B 细胞  慢性移植性肾病
英文关键词: Th cells  B cells  Chronic allograft nephropathy
基金项目:国家自然科学基金面上项目(81273267,K112409514)
作者单位
徐旭 史剑 谈春业 刘锦波 何小舟 苏州大学附属第三医院骨科实验室苏州大学附属第三医院泌尿外科实验室 
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中文摘要:
      目的:检测慢性移植性肾病(CAN)大鼠脾脏中辅助性T细胞(Th)和B细胞特征性因子表达量的变化,探究这些Th/B细胞 免疫状态在CAN病程中的作用。方法:采用Fischer-Lewis 左肾原位移植法建立大鼠慢性移植性肾病模型,Lewis-Lewis 同种自体 移植作为对照组。所有受体大鼠,术后8 周处死,取脾脏组织,进行HE 染色,拍照后采用双盲法评价脾脏组织病理变化程度及淋 巴细胞浸润情况。用Trizol 法提取脾脏组织中总RNA,采用实时荧光定量聚合酶链式反应(qRT-PCR)法检测各组脾脏中Th1, Th2,Th17,Treg 和B 细胞标志性因子的表达情况。结果:与对照组相比,CAN 大鼠脾脏出现明显的结构肿胀及淋巴细胞浸润增 多,并且Th1 细胞特征性因子IFN-γ和T-bet 表达量显著增加(P<0.001,P<0.05);Th2 细胞特征性因子GATA-3 表达升高(P<0. 001),但IL-4无变化;IFN-γ/IL-4 比例明显上调(P<0.001),T-bet/GATA3 比例没有显著差异。Th17 的特征性因子IL-17 未见明显 改变,而Treg 细胞特征性因子Foxp3 表达增加(P<0.001),IL-17/Foxp3 平衡明显向Treg 细胞偏移(P<0.05)。B 细胞激活相关因子 TNFRSF13C 和RAG1表达量均显著上调(P<0.01,P<0.05),而RAG2 水平则没有变化。结论:CAN大鼠脾脏中Th1/Th2 的活性 平衡向Th1偏移,分化平衡未出现显著变化;Th17/Treg 的平衡向Treg 细胞偏移,B细胞免疫状态也被激活,这些变化在CAN病 程的发展中起到了重要作用,并且为临床监测和治疗提供了新的依据。
英文摘要:
      Objective:To explore the immune statuses of splenic Th and B cells in a chronic allograft nephropathy (CAN) rat model, and initially investigate the potential role of them in CAN.Methods:Chronic allograft nephropathy rat model was established by orthotopic renal transplantation from Fischer to Lewis rats. Lewis to Lewis as the controls. Spleen samples were obtained 8 weeks after transplantation. Sections of spleen tissues were stained by hematoxylin-eosin (HE), and the histopathology changes were elvaluated a double-blind review. Besides, the expression of Th1/Th2/Th17/Treg/B-cell-associated immunological molecules were examined byquantitative Real-time PCR (qRT-PCR).Results:There were splenic nodules swelling and a significant increase of lymphocyte infiltration in the CAN group as compared to NC group. The expression of Th1-related molecules(IFN-γand T-bet) were increased (P<0. 001, P<0.05), while only Th2-related transcription factor GATA3 not IL-4 was enhanced (P<0.001). Similarly, IFN-γ/IL-4 rather than T-bet/GATA3 was up-regulated in the spleen with CAN (P<0.001). Moreover, there were parallel Th17-related cytokines IL-17 level but higher Foxp3 level of Treg (P<0.001), as well as lower IL-17/Foxp3 (P<0.05) in the CAN rats than NC individuals. Besides, the expression of TNFRSF13C and RAG1 which involving in B cell activation were also improved in CAN recipients (P<0.01,P<0.05).Conclusion:The changes of these Th and B cell immune statuses in the spleen are closely associated with the pathological process of CAN, which also provides a novel clue for monitoring this disease.
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