本文關(guān)鍵詞： 糖尿病視網(wǎng)膜病變 石斛多糖 炎癥因子 緊密連接蛋白 血-視網(wǎng)膜屏障　出處：《蚌埠醫(yī)學(xué)院》2016年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：研究目的近年來(lái),越來(lái)越多的研究認(rèn)為炎癥作用機(jī)制在糖尿病視網(wǎng)膜病變(diabetic retinopathy DR)的早期發(fā)展中發(fā)揮著不可或缺的作用。由此推測(cè),抗炎治療有可能成為預(yù)防DR的發(fā)展及治療提供一個(gè)新的思路。石斛作為我國(guó)傳統(tǒng)名貴中草藥之一,被認(rèn)為具有滋陰補(bǔ)腎、清肝明目的功能。多糖是石斛的主要的藥用活性成分之一,其化學(xué)成分和藥理活性作用也日益引起人們的關(guān)注,研究表明,石斛多糖具有抗炎、抗氧化、抗腫瘤等作用。本研究以糖尿病大鼠為研究對(duì)象,觀察石斛多糖與糖尿病大鼠血-視網(wǎng)膜屏障功能的關(guān)系及其具體的作用機(jī)制。實(shí)驗(yàn)方法動(dòng)物實(shí)驗(yàn):老鼠隨機(jī)分為5組,進(jìn)行實(shí)驗(yàn):(1)正常組,(2)糖尿病組,(3)糖尿病+石斛多糖低劑量組[100mg/(kg?天)],(4)糖尿病+石斛多糖中劑量組[200mg/(kg?天)],(5)糖尿病+石斛多糖高劑量組[300mg/(kg?天)]。老鼠按照55mg/kg使用鏈脲佐菌素(streptozotocin,STZ)腹腔注射,正常組組大鼠腹腔注射同等劑量檸檬酸鈉溶液,以后每周監(jiān)測(cè)血糖一次。48小時(shí)血糖高于16.7mmol/L認(rèn)為造模成功。建模成功6周后,石斛多糖各處理組進(jìn)行為期8周灌胃治療。8周后,2%戊巴比妥鈉(40mg/kg)腹腔麻醉大鼠,取所需標(biāo)本。采用蛋白印跡法(Westen Blot)檢測(cè)大鼠視網(wǎng)膜內(nèi)白介素-6(interleukin-6,IL-6)、腫瘤壞死因子-α(tumer factor-alpha,TNF-α)、血管內(nèi)皮生長(zhǎng)因子(vascular endothelial growth factor,VEGF)及視網(wǎng)膜內(nèi)緊密連接蛋白家族(ZO-1、Occludin、Claudin-5)的蛋白表達(dá)水平,酶聯(lián)免疫吸附試驗(yàn)(Enzyme Linked Immunosorbent Assay,ELISA)檢測(cè)大鼠血清、視網(wǎng)膜內(nèi)IL-6、TNF-α的含量,免疫組化觀察大鼠視網(wǎng)膜內(nèi)核轉(zhuǎn)錄因子κB(NF-κB)的表達(dá)水平,伊凡氏藍(lán)(Evans blue)染色分析糖尿病大鼠血視網(wǎng)膜屏障功能。實(shí)驗(yàn)結(jié)果與正常組比較,糖尿病組大鼠視網(wǎng)膜內(nèi)IL-6、TNF-α、VEGF的蛋白表達(dá)增加,緊密連接蛋白(ZO-1、Occludin、Claudin-5)表達(dá)水平降低,血清IL-6、TNF-α表達(dá)增加,大鼠視網(wǎng)膜滲透量增高,差異均具有統(tǒng)計(jì)學(xué)意義(P0.05)。與糖尿病組比較,石斛多糖低、中、高各劑量組大鼠視網(wǎng)膜內(nèi)IL-6、TNF-α的蛋白表達(dá)明顯降低,緊密連接蛋白(ZO-1、Occludin、Claudin-5)表達(dá)水平增高,血清IL-6、TNF-α的表達(dá)降低,石斛多糖各劑量組大鼠視網(wǎng)膜滲透量明顯降低,差異均具有統(tǒng)計(jì)學(xué)意義(P0.05)。石斛多糖各劑量組之間各指標(biāo)兩兩比較差異均無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。免疫組化切片觀察,正常大鼠視網(wǎng)膜未見(jiàn)明顯NF-κB陽(yáng)性染色,糖尿病組大鼠視網(wǎng)膜內(nèi)NF-κB表達(dá)高于正常組,與糖尿病組比較,石斛多糖各劑量組NF-κB表達(dá)降低。結(jié)論石斛多糖可以降低糖尿病視網(wǎng)膜病變大鼠體內(nèi)炎癥因子(IL-6、TNF-α)的表達(dá),進(jìn)而抑制VEGF表達(dá)上調(diào)并改善糖尿病大鼠血-視網(wǎng)膜屏障(blood-retinal barrier,BRB)的通透性。其作用機(jī)制可能與NF-κB通路有關(guān)。
[Abstract]:Research purposes in recent years. More and more studies suggest that the mechanism of inflammation in diabetic retinopathy / diabetic retinopathy DRR. Play an indispensable role in the early development of. Anti-inflammatory therapy may provide a new idea for the development and treatment of prevention Dr. As one of the traditional and precious Chinese herbal medicines, Dendrobium is considered to have nourishing yin and tonifying the kidney. Polysaccharide is one of the main medicinal active components of Dendrobium, and its chemical composition and pharmacological activity have attracted more and more attention. Studies show that Dendrobium polysaccharide has anti-inflammatory and antioxidant properties. In this study, diabetic rats were selected as the research objects. To observe the relationship between polysaccharide of Dendrobium and blood-retinal barrier function of diabetic rats and its specific mechanism. (2) Diabetic group (Dendrobium Dendrobium L.) low dosage of Dendrobium Dendrobium polysaccharide. [100 mg / kg? Dendrobium Dendrobiae (Dendrobium Dendrobium Dendrobium). [200 mg / kg? High dose group of Dendrobium Dendrobium polysaccharides. [300 mg / kg? Rats were injected intraperitoneally with streptozotocinia STZ (55 mg / kg), and rats in normal group were given the same dose of sodium citrate solution. Monitoring blood glucose once a week. 48 hours blood glucose was higher than 16.7 mmol / L, the model was successful. 6 weeks after the model was successfully established, Dendrobium polysaccharide treatment groups were treated for 8 weeks by gastric perfusion for 8 weeks. 2% pentobarbital sodium (40 mg / kg) was used to anesthetize rats intraperitoneally. Western blot was used to detect interleukin-6 interleukin-6 (IL-6) in rat retina. Tumor necrosis factor- 偽 tumer factor-alpha (TNF- 偽). Vascular endothelial growth factor (VEGF) and vascular endothelial growth factor (endothelial growth factor-VEGF-1) and tight junction protein family (ZO-1) in the retina. The protein expression level of Occludinia Claudin-5 was detected by enzyme-linked immunosorbent assay (Elisa). The content of IL-6 TNF- 偽 in serum and retina of rats was detected by ELISA.The expression of NF- 魏 B in rat retina was observed by immunohistochemistry. The function of blood-retina barrier in diabetic rats was analyzed by Evans blueblue staining. The results were compared with those in the normal group, and the results showed that IL-6 TNF- 偽 was found in the retina of diabetic rats. The protein expression of VEGF was increased, the expression level of ZO-1 Occludin-Occludin-5) was decreased, and the expression of IL-6 TNF- 偽 was increased. Compared with the diabetic group, Dendrobium polysaccharide was low, medium and high doses of IL-6 in the retina of rats. The protein expression of TNF- 偽 was significantly decreased, the expression of tight junction protein ZO-1 Occludinia Claudin-5 was increased, and the serum IL-6 was increased. The expression of TNF- 偽 was decreased, and the retinal osmotic capacity of Dendrobium polysaccharide group was significantly decreased. The difference was statistically significant (P 0.05). There was no significant difference in each index between different dose groups of Dendrobium officinale. The expression of NF- 魏 B in retina of diabetic group was higher than that of normal group, which was higher than that of diabetic group. Conclusion Dendrobium polysaccharide can reduce the expression of IL-6 TNF- 偽 in diabetic retinopathy rats. And then inhibit the up-regulation of VEGF expression and improve the blood-retinal barrier blood-retinal barrier in diabetic rats. The mechanism of BRBs permeability may be related to NF- 魏 B pathway.
【學(xué)位授予單位】：蚌埠醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R276.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 劉巨平;李筱榮;;糖尿病視網(wǎng)膜病變:一種非可控性炎癥[J];中華實(shí)驗(yàn)眼科雜志;2014年01期

2 柴金珍;黃娟萍;劉靜;梁錦杰;魯湘鄂;朱盛山;;不同石斛的藥理作用研究現(xiàn)狀[J];中成藥;2013年12期

3 余文珍;王曉寧;施紅;;石斛合劑對(duì)衰老糖尿病大鼠胰腺P53表達(dá)的影響[J];中華中醫(yī)藥學(xué)刊;2012年11期

4 凌志揚(yáng);房玉良;;石斛的化學(xué)成分及藥理作用[J];中國(guó)當(dāng)代醫(yī)藥;2012年05期

5 楊仲;趙敏;王明奎;喻凱;;迭鞘石斛所含多糖對(duì)小鼠免疫細(xì)胞影響的研究[J];安徽農(nóng)業(yè)科學(xué);2011年22期

6 尚喜雨;;多糖在不同來(lái)源不同部位鐵皮石斛中的分布規(guī)律研究[J];中國(guó)現(xiàn)代藥物應(yīng)用;2010年13期

7 李小瓊;金徽;葛曉軍;唐彥萍;;金釵石斛多糖對(duì)脂多糖誘導(dǎo)的小鼠腹腔巨噬細(xì)胞分泌TNF-α·NO的影響[J];安徽農(nóng)業(yè)科學(xué);2009年28期

，

本文編號(hào)：1486627