Apelin-13对大鼠实验性自身免疫性神经炎的防治作用及其机制

发布时间：2018-04-28 16:47

本文选题：Apelin-13 + 实验性自身免疫性神经炎　；参考：《中南大学》2014年硕士论文

【摘要】：目的观察Apelin-13对大鼠实验性自身免疫性神经炎的防治作用并探讨其可能的机制。方法采用周围神经髓鞘抗原(P257-81)注射入Lewis大鼠后肢足垫诱导EAN模型。Lewis雄性大鼠随机分为对照组、EAN模型组和Apelin-13处理组。Apelin-13处理组于免疫当天至第15天每天尾静脉注射Apelin-13(0.1mg/kg)。观察各组大鼠发病情况和坐骨神经组织病理学变化及炎症细胞浸润情况。制备脾脏单个淋巴细胞悬液,加入刀豆蛋白A、脂多糖和周围神经髓鞘抗原(P257-81)不同刺激物孵育48小时后,采用流式细胞术检测淋巴细胞增殖。采用酶联免疫(ELISA)法检测大鼠血浆中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、干扰素-γ (IFN-γ)、白细胞介素-4(IL-4)和转化生长因子-β(TGF-β)的水平,采用实时定量PCR和Western blot检测淋巴结中TNF-α、IL-6、IFN-γ、IL-4和TGF-β mRNA和蛋白的表达。结果(1)EAN模型组大鼠坐骨神经神经束间和神经束内有大量的炎症细胞浸润,存在局灶性脱髓鞘现象。与对照组比较,EAN模型组脾脏淋巴细胞的基础增殖以及LPS和周围神经髓鞘抗原(P257-81)诱导的细胞增殖水平均显著增高(均P0.05)。与对照组比较,EAN模型组大鼠血浆中TNF-α、IL-6和IFN-γ水平显著升高,而血浆中IL-4和TGF-β的水平显著降低(均P0.05)。与对照组比较,EAN模型组大鼠淋巴结中TNF-α、IL-6和IFN-γ mRNA和蛋白的表达显著升高,而IL-4和TGF-β mRNA和蛋白的表达显著降低(均P0.05)。(2)与EAN模型组比较,Apelin-13处理组大鼠最初发病时间明显延长,高峰期临床评分显著降低(P0.05),坐骨神经神经束间和神经束内炎症细胞浸润明显减少,脱髓鞘现象明显减少。与EAN模型组相比,Apelin-13处理组的基础细胞增殖以及LPS和周围神经髓鞘抗原(P257-81)诱导的细胞增殖水平均显著降低(均P0.05)。与EAN模型组比较,Apelin-13处理组大鼠血浆中TNF-α、IL-6和IFN-γ水平显著降低,而血浆中IL-4和TGF-β的水平显著增加(均P0.05)。与对照组比较,Apelin-13处理组大鼠淋巴结中TNF-α、IL-6和IFN-γ mRNA和蛋白的表达显著降低,而IL-4和TGF-β mRNA和蛋白的表达显著升高(均P0.05)。结论Apelin-13对实验性自身免疫性神经炎大鼠有防治作用,其机制可能与下调TNF-α、IL-6和IFN-γ的表达和上调IL-4和TGF-β有关。图11幅,表8个,参考文献73篇。
[Abstract]:Objective to observe the preventive and therapeutic effects of Apelin-13 on experimental autoimmune neuritis in rats and to explore its possible mechanism. Methods Lewis rats were injected with peripheral nerve myelin sheath antigen (P257-81) to induce EAN model. The male rats were randomly divided into control group and Apelin-13 treatment group. Apelin-13 group was injected with 0.1 mg / kg Apelin-13 in caudal vein every day from the day of immunization to the 15th day. The pathological changes of sciatic nerve and inflammatory cell infiltration were observed. Spleen single lymphocyte suspension was incubated with concanavalin A, lipopolysaccharide (LPS) and peripheral nerve myelin antigen (P257-81) for 48 hours. Lymphocyte proliferation was detected by flow cytometry. The levels of TNF- 伪, IL-6, IFN- 纬, IL-4IL-4 and TGF- 尾 in plasma of rats were detected by enzyme linked immunosorbent assay (Elisa). Real-time quantitative PCR and Western blot were used to detect the expression of IL-4, TGF- 尾 mRNA and protein in lymph nodes. Results there were a large number of inflammatory cells infiltrating between the sciatic nerve bundles and the nerve bundles in the EAN group, and there was focal demyelination. Compared with the control group, the basic proliferation of spleen lymphocytes and the level of proliferation induced by LPS and P257-81 were significantly increased in EAN model group (all P 0.05). Compared with the control group, the plasma levels of TNF- 伪, IL-6 and IFN- 纬 in the EAN group were significantly increased, while the levels of IL-4 and TGF- 尾 in the plasma were significantly decreased (P 0.05). Compared with the control group, the expression of TNF- 伪 IL-6 and IFN- 纬 mRNA and protein in the lymph nodes of rats in the EAN group was significantly increased, while the expression of IL-4 and TGF- 尾 mRNA and protein was significantly decreased in the EAN model group (all P0.05A. 2) compared with the EAN model group, the initial onset time in the Apelin-13 group was significantly longer than that in the EAN model group. The peak clinical score decreased significantly (P 0.05), inflammatory cell infiltration and demyelination of sciatic nerve bundles and nerve bundles decreased significantly. Compared with the EAN model group, the basal cell proliferation and the proliferation level induced by LPS and P257-81 were significantly decreased in the Apelin-13 treatment group (all P 0.05). Compared with the EAN model group, the plasma levels of TNF- 伪, IL-6 and IFN- 纬 in the Apelin-13 group were significantly decreased, while the levels of IL-4 and TGF- 尾 in the plasma were significantly increased (P 0.05). Compared with the control group, the expression of TNF- 伪 mRNA and IFN- 纬 mRNA and protein in the lymph nodes of rats treated with Apelin-13 was significantly decreased, while the expression of IL-4 and TGF- 尾 mRNA and protein were significantly increased (all P 0.05). Conclusion Apelin-13 has preventive and therapeutic effects on experimental autoimmune neuritis in rats. The mechanism may be related to down-regulation of TNF- 伪 IL-6 and IFN- 纬 expression and up-regulation of IL-4 and TGF- 尾. There are 11 figures, 8 tables and 73 references.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R745

【参考文献】