本文選題：氧化應(yīng)激 + 慢性間歇性低氧��； 參考：《桂林醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:研究銀杏葉提取物(extract ginkgo biloba,EGB)通過Nrf2-ARE信號(hào)通路對(duì)慢性間歇性低氧(chronic intermittent hypoxia,CIH)大鼠胰島β細(xì)胞凋亡中的作用及干預(yù)作用。方法:建立慢性間歇性低氧大鼠模型,將SD雄性大鼠36只隨機(jī)分成兩組,常氧對(duì)照組為12只,慢性間歇性低氧模型組為24只;第12周造模結(jié)束后,提取各組原代胰島β細(xì)胞;常氧對(duì)照組細(xì)胞正常培養(yǎng);慢性間歇性低氧模型組β細(xì)胞根據(jù)處理因素不同分為慢性間歇性低氧模型對(duì)照組(OSAS對(duì)照組)、抗Nrf2抗體組、Nrf2-ARE通路激活劑組、銀杏葉提取物低、中、高劑量組共6組,其余組予以等量平衡鹽溶液,加入含有不同處理藥物的培養(yǎng)基相同條件培養(yǎng)。鑒定胰島β細(xì)胞使用DTZ染色法,使用ELISA法檢測(cè),各組胰島β細(xì)胞及上清液中谷胱甘肽前體(GSH-PX)、丙二醛(MAD)和活性氧(ROS)、腫瘤壞死因子(TNF-a)、白介素-8(IL-8)、8-異前列腺素(8-ISO)、超化物歧化酶(SOD)的含量,采用流式細(xì)胞儀對(duì)胰島β細(xì)胞凋亡進(jìn)行檢測(cè),使用Westerm-Blot法測(cè)定胰島β細(xì)胞中核因子2相關(guān)因子(Nrf2)、血紅素加氧酶-1(HO-1)、醌氧化還原酶(NQO1)、r-谷氨酰半胱氨酸合成酶(r-GCS)的表達(dá)水平。結(jié)果:與常氧對(duì)照組對(duì)比,OSAS對(duì)照組胰島β細(xì)胞中ROS及上清液中MDA、TNF-a、IL-8、8-ISO水平均上升,GSH-PX、SOD表達(dá)下降,胰島β細(xì)胞凋亡率升高,Nrf2表達(dá)升高,HO-1、NQO1、r-GCS在胰島β細(xì)胞表達(dá)水平均下調(diào),差異有統(tǒng)計(jì)學(xué)意義;與OSAS對(duì)照組對(duì)比,抗Nrf2抗體組胰島β細(xì)胞中ROS及上清液中MDA、IL-8、TNF-a、8-ISO水平升高及SOD、GSH-PX下降更顯著,胰島β細(xì)胞凋亡率升高,HO-1、NQO1表達(dá)水平均降低,差異有統(tǒng)計(jì)學(xué)意義,而Nrf2、r-GCS表達(dá)水平無明顯變化(P0.05);Nrf2-ARE通路激活劑組胰島β細(xì)胞中ROS及上清液中MDA、IL-8、TNF-a、8-ISO表達(dá)水平均降低,SOD及GSH-PX表達(dá)水平均上升,胰島β細(xì)胞凋亡率下降,Nrf2、HO-1、NQO1、r-GCS在胰島β細(xì)胞表達(dá)均升高,差異有統(tǒng)計(jì)學(xué)意義;銀杏葉提取物低劑量組胰島β細(xì)胞中ROS及上清液中IL-8、8-ISO水平均下降,SOD表達(dá)水平均上升,差異有統(tǒng)計(jì)學(xué)意義;TNF-a、GSH-PX、MDA無差異(P0.05);胰島β細(xì)胞凋亡率下降,Nrf2、HO-1、r-GCS在胰島β細(xì)胞表達(dá)均升高,NQO1表達(dá)水平下調(diào),差異有統(tǒng)計(jì)學(xué)意義;銀杏葉提取物中、高劑量組胰島β細(xì)胞上清液MDA、ROS、IL-8、TNF-a、8-ISO表達(dá)水平均降低,SOD及GSH-PX表達(dá)水平均上升,胰島β細(xì)胞凋亡率下降,Nrf2、HO-1、NQO1、r-GCS在胰島β細(xì)胞表達(dá)均升高,且高濃度組比低濃度組變化更明顯,差異有統(tǒng)計(jì)學(xué)意義。結(jié)論:Nrf2-ARE信號(hào)通路能調(diào)節(jié)間歇性低氧所致的氧化應(yīng)激及炎癥因子水平,減少胰島β細(xì)胞的凋亡,對(duì)胰島β細(xì)胞起保護(hù)作用。銀杏葉提取物通過Nrf2-ARE信號(hào)通路,調(diào)節(jié)間歇性低氧誘導(dǎo)胰島β細(xì)胞凋亡中的氧化物質(zhì)及炎癥因子的水平,減少胰島β細(xì)胞凋亡率,對(duì)胰島β細(xì)胞有保護(hù)作用。
[Abstract]:Aim: to study the effect of (extract ginkgo biloba (EGB) on apoptosis of islet 尾 cells in rats with chronic intermittent hypoxia (chronic intermittent hypoxia via Nrf2-ARE signaling pathway. Methods: the chronic intermittent hypoxic rat model was established and 36 SD male rats were randomly divided into two groups: 12 rats in the normoxic control group and 24 rats in the chronic intermittent hypoxia model group. The 尾 cells of chronic intermittent hypoxia model group were divided into chronic intermittent hypoxia model control group (OSAS control group), anti-Nrf2 antibody group with Nrf2-ARE pathway activator group, ginkgo biloba leaf extract low, medium, according to the treatment factors, chronic intermittent hypoxia model group 尾 cells were divided into chronic intermittent hypoxia model control group (OSAS control group), anti-Nrf2 antibody group and Nrf2-ARE pathway activator group. The other groups were treated with the same amount of equilibrium salt solution and cultured in the same culture medium containing different treatment drugs. Islet 尾 cells were identified by DTZ staining and Elisa. The contents of glutathione precursor (GSH-PX), malondialdehyde (mad) and reactive oxygen species (Ros), tumor necrosis factor (TNF-a), interleukin-8 (IL-8) 8-isoprostaglandin (8-ISO), supernatant dismutase (SOD) in islet 尾 cells and supernatant dismutase (SOD) were detected by Elisa. Apoptosis of islet 尾 cells was detected by flow cytometry. The expression levels of nuclear factor-2 (Nrf2), heme oxygenase-1 (HO-1) and quinone redox enzyme (NQO1) r-glutamylcysteine synthase (r-GCS) in islet 尾 cells were determined by Westerm-Blot method. Results: the levels of Ros in islet 尾 cells and the levels of MDA-TNF-aIL-8 8-ISO in OSAS control group were increased and the expression of GSH-PXO SOD was decreased, and the expression of HO-1NQO1r-GCS was down-regulated in islet 尾 cells compared with normoxic control group. Compared with OSAS control group, the levels of Ros in islet 尾 cells in anti-Nrf2 antibody group and MDA-8 IL-8, TNF-aF- 8-ISO in supernatant, and the decrease of GSH-PX in islet 尾 cells were significantly higher than those in OSAS control group, and the expression level of HO-1NQO1 in islet 尾 -cell apoptosis was significantly lower than that in OSAS control group. The expression of Ros in islet 尾 cells and the expression of MDA-IL-8, TNF-aF- 8-ISO in pancreatic islet 尾 cells in Nrf2-ARE pathway activator group were decreased, and the expression of SOD and GSH-PX were increased. The apoptotic rate of islet 尾 cells was decreased, and the expression of NQO1O1r-GCS in islet 尾 cells was increased, but the expression of Nrf2-ARE pathway activator group was higher than that of Nrf2-ARE pathway activator group, while the expression of SOD and GSH-PX in pancreatic islet 尾 cells were increased in Nrf2-ARE pathway activator group. The levels of Ros in islet 尾 cells and IL-88-ISO in supernatant of ginkgo biloba extract decreased significantly. There was no significant difference in MDA between TNF-a and GSH-PXN (P0.05), the decrease of apoptosis rate of islet 尾 cells and the down-regulation of the expression of NQO1 in islet 尾 cells by Nrf2HO-1HO-1r-GCS, there was significant difference in the expression of NQO1 in ginkgo biloba leaves. In high dose group, the expression of MDA-ROSS-IL-8 and TNF-aO8-ISO in the supernatant of islet 尾 cells decreased, and the expression of SOD and GSH-PX increased, and the apoptosis rate of islet 尾 cells decreased. The expression of Nrf2HO-1NQO1O1r-GCS in islet 尾 cells increased, and the changes in high concentration group were more obvious than those in low concentration group. The difference is statistically significant. Conclusion the fraction of Nrf2-ARE signaling pathway can regulate oxidative stress and inflammatory factor level induced by intermittent hypoxia, reduce the apoptosis of islet 尾 cells, and play a protective role on islet 尾 cells. Through Nrf2-ARE signaling pathway, ginkgo biloba extract regulates the levels of oxidants and inflammatory factors in the apoptosis of islet 尾 cells induced by intermittent hypoxia, reduces the apoptosis rate of islet 尾 cells, and has protective effect on islet 尾 cells.
【學(xué)位授予單位】：桂林醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R766

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本文編號(hào)：2106027