發(fā)布時(shí)間：2018-04-28 16:47

  本文選題：Apelin-13 + 實(shí)驗(yàn)性自身免疫性神經(jīng)炎��； 參考：《中南大學(xué)》2014年碩士論文

【摘要】：目的觀察Apelin-13對(duì)大鼠實(shí)驗(yàn)性自身免疫性神經(jīng)炎的防治作用并探討其可能的機(jī)制。 方法采用周圍神經(jīng)髓鞘抗原(P257-81)注射入Lewis大鼠后肢足墊誘導(dǎo)EAN模型。Lewis雄性大鼠隨機(jī)分為對(duì)照組、EAN模型組和Apelin-13處理組。Apelin-13處理組于免疫當(dāng)天至第15天每天尾靜脈注射Apelin-13(0.1mg/kg)。觀察各組大鼠發(fā)病情況和坐骨神經(jīng)組織病理學(xué)變化及炎癥細(xì)胞浸潤(rùn)情況。制備脾臟單個(gè)淋巴細(xì)胞懸液,加入刀豆蛋白A、脂多糖和周圍神經(jīng)髓鞘抗原(P257-81)不同刺激物孵育48小時(shí)后,采用流式細(xì)胞術(shù)檢測(cè)淋巴細(xì)胞增殖。采用酶聯(lián)免疫(ELISA)法檢測(cè)大鼠血漿中腫瘤壞死因子-α(TNF-α)、白細(xì)胞介素-6(IL-6)、干擾素-γ (IFN-γ)、白細(xì)胞介素-4(IL-4)和轉(zhuǎn)化生長(zhǎng)因子-β(TGF-β)的水平,采用實(shí)時(shí)定量PCR和Western blot檢測(cè)淋巴結(jié)中TNF-α、IL-6、IFN-γ、IL-4和TGF-β mRNA和蛋白的表達(dá)。 結(jié)果(1)EAN模型組大鼠坐骨神經(jīng)神經(jīng)束間和神經(jīng)束內(nèi)有大量的炎癥細(xì)胞浸潤(rùn),存在局灶性脫髓鞘現(xiàn)象。與對(duì)照組比較,EAN模型組脾臟淋巴細(xì)胞的基礎(chǔ)增殖以及LPS和周圍神經(jīng)髓鞘抗原(P257-81)誘導(dǎo)的細(xì)胞增殖水平均顯著增高(均P0.05)。與對(duì)照組比較,EAN模型組大鼠血漿中TNF-α、IL-6和IFN-γ水平顯著升高,而血漿中IL-4和TGF-β的水平顯著降低(均P0.05)。與對(duì)照組比較,EAN模型組大鼠淋巴結(jié)中TNF-α、IL-6和IFN-γ mRNA和蛋白的表達(dá)顯著升高,而IL-4和TGF-β mRNA和蛋白的表達(dá)顯著降低(均P0.05)。(2)與EAN模型組比較,Apelin-13處理組大鼠最初發(fā)病時(shí)間明顯延長(zhǎng),高峰期臨床評(píng)分顯著降低(P0.05),坐骨神經(jīng)神經(jīng)束間和神經(jīng)束內(nèi)炎癥細(xì)胞浸潤(rùn)明顯減少,脫髓鞘現(xiàn)象明顯減少。與EAN模型組相比,Apelin-13處理組的基礎(chǔ)細(xì)胞增殖以及LPS和周圍神經(jīng)髓鞘抗原(P257-81)誘導(dǎo)的細(xì)胞增殖水平均顯著降低(均P0.05)。與EAN模型組比較,Apelin-13處理組大鼠血漿中TNF-α、IL-6和IFN-γ水平顯著降低,而血漿中IL-4和TGF-β的水平顯著增加(均P0.05)。與對(duì)照組比較,Apelin-13處理組大鼠淋巴結(jié)中TNF-α、IL-6和IFN-γ mRNA和蛋白的表達(dá)顯著降低,而IL-4和TGF-β mRNA和蛋白的表達(dá)顯著升高(均P0.05)。 結(jié)論Apelin-13對(duì)實(shí)驗(yàn)性自身免疫性神經(jīng)炎大鼠有防治作用,其機(jī)制可能與下調(diào)TNF-α、IL-6和IFN-γ的表達(dá)和上調(diào)IL-4和TGF-β有關(guān)。圖11幅,表8個(gè),參考文獻(xiàn)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- 偽, IL-6, IFN- 緯, 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- 偽, IL-6 and IFN- 緯 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- 偽 IL-6 and IFN- 緯 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- 偽, IL-6 and IFN- 緯 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- 偽 mRNA and IFN- 緯 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- 偽 IL-6 and IFN- 緯 expression and up-regulation of IL-4 and TGF- 尾. There are 11 figures, 8 tables and 73 references.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R745

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