本文關(guān)鍵詞：芍藥苷抑制糖尿病視網(wǎng)膜病變基質(zhì)金屬蛋白酶9激活的研究　出處：《南京醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 糖尿病視網(wǎng)膜病變 基質(zhì)金屬蛋白酶9 小膠質(zhì)細(xì)胞 芍藥苷

【摘要】：背景:糖尿病視網(wǎng)膜病變(DR)是糖尿病引起的并發(fā)癥之一,是導(dǎo)致發(fā)達(dá)國家糖尿病患者后天視力喪失的主要原因。其病理表現(xiàn)為:視網(wǎng)膜細(xì)胞凋亡、視網(wǎng)膜神經(jīng)功能紊亂、新生血管的大量生成等。盡管對這種病變的研究眾多,但迄今對這種多因素疾病的確切機(jī)制還不清楚。小膠質(zhì)細(xì)胞在DR疾病發(fā)展過程發(fā)揮著重要作用�；|(zhì)金屬蛋白酶(MMPs)能夠降解細(xì)胞外基質(zhì),調(diào)節(jié)細(xì)胞內(nèi)穩(wěn)態(tài)平衡�；|(zhì)金屬蛋白9(MMP-9)損害血視網(wǎng)膜屏障(BRB),與DR的發(fā)展過程密切相關(guān)。芍藥苷,屬于單萜苷化合物,對小膠質(zhì)細(xì)胞具有強(qiáng)大的免疫調(diào)節(jié)作用。本文我們研究芍藥苷是否可通過抑制高糖刺激的小膠質(zhì)細(xì)胞MMP-9的激活,從而改善DR。目的:研究芍藥苷對糖尿病視網(wǎng)膜病變MMP-9激活的作用。實(shí)驗(yàn)方法:高糖刺激小膠質(zhì)細(xì)胞,模擬體內(nèi)糖尿病模型;腹腔注射STZ(鏈脲佐菌素)建立糖尿病小鼠模型;明膠酶譜檢測MMP-9的活性;免疫印記法檢測信號通路蛋白水平的表達(dá);免疫熒光法檢測NF-κB核轉(zhuǎn)錄情況;血糖儀測定小鼠血糖水平;HE染色觀察小鼠視網(wǎng)膜形態(tài)結(jié)構(gòu)。實(shí)驗(yàn)結(jié)果:高糖能夠引起小膠質(zhì)細(xì)胞MMP-9的激活,這種作用可被高遷移率族蛋白1(HMGB1)抑制劑、Toll樣受體4(TLR-4)抑制劑、p38絲裂素活化蛋白激酶(p38MAPK)抑制劑、核轉(zhuǎn)錄因子-κB(NF-κB)抑制劑所取消。TLR-4抑制劑減少高糖誘導(dǎo)的小膠質(zhì)細(xì)胞p38MAPK磷酸化水平。芍藥苷增加熱休克蛋白(HSP70)和細(xì)胞因子信號傳導(dǎo)抑制蛋白-3(SOCS3)蛋白水平的表達(dá),減少小膠質(zhì)細(xì)胞MMP-9的表達(dá)。芍藥苷誘發(fā)的SOCS3蛋白的表達(dá)被TLR-4抑制劑所取消。在STZ誘導(dǎo)的糖尿病鼠中,芍藥苷能夠誘發(fā)SOCS3蛋白表達(dá),減少M(fèi)MP-9激活。芍藥苷抑制STZ誘導(dǎo)的IBA1,IL-1β表達(dá),減少STZ誘導(dǎo)的糖尿病鼠血糖水平的升高,改善STZ誘導(dǎo)的糖尿病鼠視網(wǎng)膜的形態(tài)結(jié)構(gòu)的改變。結(jié)論:高糖通過激活HMGB1-TLR4-NF-κB信號通路引起小膠質(zhì)細(xì)胞MMP-9的激活,引起DR。芍藥苷通過HSP70/TLR4/NF-κB信號通路誘發(fā)SOCS3表達(dá),減少M(fèi)MP-9激活,改善DR。
[Abstract]:Background: diabetic retinopathy (DR) is one of the complications caused by diabetes and is the main cause of the loss of acquired visual acuity in patients with diabetes in developed countries. The pathological features are retinal cell apoptosis, retinal nerve dysfunction, and a large number of neovascularization. Although there are many studies on this disease, the exact mechanism of this multifactor disease is not yet clear. Microglia play an important role in the development of DR disease. Matrix metalloproteinases (MMPs) can degrade extracellular matrix and regulate homeostasis in cells. Matrix metalloprotein 9 (MMP-9) damages the blood retinal barrier (BRB), which is closely related to the development of DR. Paeoniflorin, a monoterpene glycoside, has a strong immunomodulatory effect on microglia. In this article, we study whether paeoniflorin can improve the activation of MMP-9 in microglia stimulated by high glucose, thus improving DR. Objective: To study the effect of paeoniflorin on the activation of MMP-9 in diabetic retinopathy. Methods: high glucose stimulated microglia, simulated in vivo model of diabetes; intraperitoneal injection of STZ (STZ) to establish diabetic mouse model; gelatinase MMP-9 activity was detected; to detect the expression of signal transduction protein level by Western blot detection; NF- kappa B transcription by immunofluorescence method; Determination of blood glucose levels in mice blood glucose meter observation on the morphology of mouse retina; HE staining. Results: high glucose could induce the activation of microglia MMP-9, this effect was of high mobility group protein 1 (HMGB1) inhibitors, Toll like receptor 4 (TLR-4) inhibitor, p38 mitogen activated protein kinase (p38MAPK) inhibitor, nuclear factor kappa B (NF- K B) inhibitor of the cancelled. TLR-4 inhibitors reduce the level of p38MAPK phosphorylation in high glucose induced microglia. Paeoniflorin increases the expression of heat shock protein (HSP70) and cytokine signal transduction inhibitor protein -3 (SOCS3) protein, and reduces the expression of MMP-9 in microglia. The expression of paeoniflorin induced SOCS3 protein was cancelled by TLR-4 inhibitor. In STZ induced diabetic rats, paeoniflorin can induce the expression of SOCS3 protein and reduce the activation of MMP-9. Paeoniflorin inhibited the expression of IBA1 and IL-1 beta induced by STZ, reduced the increase of blood glucose level in diabetic rats induced by STZ, and improved the morphological structure of retina in STZ induced diabetic rats. Conclusion: high glucose induces the activation of MMP-9 in microglia by activating the HMGB1-TLR4-NF- kappa B signaling pathway, causing DR. Paeoniflorin induces SOCS3 expression through HSP70/TLR4/NF- kappa B signaling pathway, reducing MMP-9 activation and improving DR.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R285

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