【作者】 张翔；

【导师】 耿燕；

【作者基本信息】 青岛大学 ， 眼科学， 2009， 硕士

【摘要】 目的从体外细胞培养方面,观察槲皮素(quercetin,QUE)对高糖状态下牛视网膜毛细血管周细胞(bovine retinal microvascular pericytes,BRPCs)增殖和凋亡的作用,以及对BRPCs氧化应激状态的影响,并探讨QUE对糖尿病视网膜微血管的保护作用。方法原代培养BRPCs,使用免疫组化法进行鉴定。利用25mmol/L葡萄糖模拟高糖状态,同时分别将25、50、100μmol/LQUE作用于高糖培养的BRPCs,并进行下列实验:采用MTT比色法检测不同处理时间(2d、4d、6d)QUE对高糖状态下BRPCs增殖的影响;高糖培养6天后,TUNEL法检测QUE对高糖状态下BRPCs凋亡的影响;比色法测定QUE对高糖状态下BRPCs超氧化物歧化酶活力(superoxidedismutase,SOD)活力和丙二醛(malondialdehyde,MDA)含量的影响。结果获得较纯净的BRPCs,并能连续传代。MTT比色法检测显示:高糖组较正常组吸光度值降低(P＜0.01),QUE组较高糖组吸光度值升高(P＜0.05),以高浓度QUE组吸光度值最高;TUNEL法检测显示:高糖状态下部分BRPCs出现凋亡改变,高糖组较正常组凋亡率升高(P＜0.01),QUE组较高糖组凋亡率降低(P＜0.05),以高浓度QUE凋亡率最低:比色法测定显示:高糖组较正常组SOD活力降低(P＜0.01)、MDA含量升高(P＜0.01),QUE组较高糖组SOD活力升高(P＜0.05)、MDA含量减低(P＜0.05),以高浓度QUE组最显著。结论高糖对体外培养BRPCs的增殖有显著的抑制作用,并随着时间的延长而加重;QUE可减轻高糖对BRPCs的氧化应激损伤,抑制BRPCs凋亡,促进BRPCs增殖,使高糖环境中BRPCs中SOD活性升高,MDA含量降低,对高糖培养的BRPCs具有良好的保护作用。更多还原

【Abstract】 Objective To investigate the effects of quercetin on the proliferation,apoptosis and oxidative stress status of bovine retinal microvascular pericytes(BRPCs) cultured with the high glucose medium in vitro,to explore the protection of quercetin on retinal microvascular of diabetic retinopathy.Methods BRPCs were primary cultured and assayed with immunohistochemistry.Using 25mmol/L glucose to simulate high glucose condition,BRPCs were cultured with the high glucose medium and incubated with different concentrations of quercetin(25,50, 100μmol/L) in the following experiments.Proliferation of BRPCs was assessed by MTT colorimetry at different time(2,4 and 6 days).Cell apoptosis of BRPCs was detected by TUNEL straining after 6 days.The activity of superoxide dismutase(SOD) and the content of malondialdehyde(MDA) in BRPCs were determined by colorimetry.Results Relative pure BRPCs were obtained and stably passaged.MTT assay showed that: compared with normal group,the absorbance value reduced in high glucose group (P<0.01);compared with high glucose group,the absorbance value increased in quercetin group(P<0.05);the absorbance value was highest in high concentration quercetin group. TUNEL assay showed that:a few cells appeared apoptosis;compared with normal group, the apoptosis rate increased in high glucose group(P<0.01);compared with high glucose group,the apoptosis rate reduced in quercetin group(P<0.05);the apoptosis rate was lowest in high concentration quercetin group.Colorimetry showed that:compared with normal group,the activity of SOD reduced and the content of MDA increased in high glucose group(P<0.01);compared with high glucose group,the activity of SOD increased and the content of MDA reduced in quercetin group(P<0.05);the activity of SOD was highest and the content of MDA was lowest in high concentration quercetin group.Conclusion High glucose has an obvious inhibitory effect on the proliferation of BRPCs and the inhibition increase with culturing time.Quercetin can alleviate the damage of oxidative stress on BRPCs in high glucose condition,inhibit apoptosis,promote proliferation,increase the activity of SOD and decrease the content of MDA.Quercetin possesses significant protective effect on BRPCs cultured with the high glucose medium in vitro.更多还原