痛风小鼠模型的建立及白藜芦醇对其干预作用

发布时间：2018-06-05 03:01

本文选题：Sirt1 + 白藜芦醇　；参考：《复旦大学》2013年硕士论文

【摘要】：研究背景与目的痛风是嘌呤代谢紊乱和(或)尿酸排泄障碍导致血尿酸升高的一组疾病,以在关节滑液和其他组织中形成单钠尿酸盐(monosodium urate, MSU)晶体为特点。痛风性关节炎以反复发作的急性炎症、慢性痛风石形成及骨侵蚀为主要临床表现,高尿酸血症是其重要的病理基础。目前研究认为痛风性关节炎的机制主要是MSU晶体通过识别细胞表面的Toll样受体激活NF-κB、AP-1信号通路启动炎症。MSU晶体也能够激活NALP3炎症复合体,最终导致成熟的IL-1β的释放。因此降低血尿酸浓度、抑制炎症通路可能是预防及治疗痛风发作的有效措施。近期研究发现,沉默调节因子1(silent information regulator1, Sirtl)可能通过抑制NF-κB和AP-1的转录活性以及下调多种炎症相关因子的表达来缓解炎症。此外,Sirt1可调节细胞代谢,参与调控多种代谢性疾病,研究证明Sirt1激动剂可降低高血糖并抑制肝脏脂肪变性。白藜芦醇(resveratrol, Res)为Sirt1的激动剂之一,是一种多酚类天然植物抗毒素。本文通过建立持续的小鼠高尿酸血症模型及MSU诱导的小鼠急性痛风性关节炎模型,给予Sirt1激动剂白藜芦醇干预各模型,观察白藜芦醇在痛风中的作用。研究内容与方法一、小鼠持续高尿酸血症模型的建立及白藜芦醇对小鼠高尿酸血症的干预作用以含25%酵母膏饲料喂食6周龄雄性昆明小鼠,联合氧嗪酸钾250mg/Kg腹腔注射,每天一次,连续3周建立高尿酸血症模型。治疗组在建模的同时予Sirt1激动剂白藜芦醇20mg/Kg灌胃,每天一次,不同时间点经内眦静脉采血检测小鼠血尿酸、肌酐浓度,HE染色法检测肾脏组织的病理改变。二、白藜芦醇对MSU诱导的小鼠急性关节炎的干预在6-8周龄的C57BL/6雄性小鼠足掌部注射MSU诱导痛风关节炎模型,通过数显卡尺测量足掌厚度评价关节肿胀程度,应用检测99mTc吸收率评价关节炎症程度,于不同时间点给予不同剂量的白藜芦醇干预治疗,评估关节炎症变化。研究结果酵母膏联合氧嗪酸钾可建立持续、稳定的高尿酸血症模型,其中建模的第3-14d血尿酸水平显著升高且无肾功能损害表现,而建模的第21d血尿酸水平上升幅度减缓,肌酐轻度上升,肾脏病理见肾小球囊壁增厚,肾间质炎性细胞浸润,肾小管上皮细胞肿胀。干预组在诱导高尿酸模型同时给予白藜芦醇治疗,小鼠血尿酸水平较模型组同期明显下降(P0.05),且上述肾脏病变好转。 MSU晶体局部注射可诱导小鼠急性关节炎,炎症表现程度与MSU浓度呈正比,具有一定的时间限定性,在MSU注射后8-12h小鼠足掌炎症表现最显著,24小时后炎症逐渐减轻。给予白藜芦醇预处理后MSU诱导的关节炎症较对照组有所缓解(P0.05),白藜芦醇分别预处理3天、7天、14天后再予MSU诱导炎症发现预处理14天组小鼠关节炎症最轻,且该作用与白藜芦醇的剂量呈正相关。研究结论 1.酵母膏联合氧嗪酸钾可建立持续、稳定的高尿酸血症模型。 2. Sirt1激动剂白藜芦醇可以通过其降尿酸作用和炎症调节作用来预防和治疗痛风发作。
[Abstract]:Research background and purpose
Gout is a group of diseases characterized by a disorder of the purine metabolism and / or uric acid excretion, which is characterized by the formation of monosodium urate (MSU) crystals in synovial fluid and other tissues. The main clinical manifestations of gouty arthritis are recurrent acute inflammation, chronic gout formation and bone erosion, and high uric acid The main mechanism of hyperarthritis is that the mechanism of gouty arthritis is that the MSU crystal activates NF- kappa B by identifying the Toll like receptor on the surface of the cell, and the AP-1 signaling pathway activates the NALP3 inflammatory complex and eventually leads to the release of the mature IL-1 beta, thus reducing the concentration of blood uric acid and inhibiting the inflammation. The disease pathway may be an effective measure to prevent and treat gout attacks.
Recent studies have found that silent information regulator1 (Sirtl) may alleviate inflammation by inhibiting the transcriptional activity of NF- kappa B and AP-1 and down regulating the expression of a variety of inflammatory related factors. In addition, Sirt1 can regulate cell metabolism and participate in the regulation of various metabolic diseases. Studies have shown that Sirt1 agonists can reduce hyperglycemia and Inhibition of hepatic steatosis, resveratrol (Res) is one of the agonists of Sirt1, and is a polyphenol natural plant antitoxin.
By establishing a continuous hyperuricemia model in mice and the model of acute gouty arthritis in mice induced by MSU, the effects of resveratrol on the gout were given by the intervention of resveratrol in each model with Sirt1 agonist.
Research content and method
Establishment of a mouse model of persistent hyperuricemia and intervention of resveratrol on hyperuricemia in mice
6 weeks of male Kunming mice were fed with 25% yeast extract, combined with potassium oxazine 250mg/Kg intraperitoneally, once a day for 3 weeks to establish hyperuricemia model. The treatment group was given Sirt1 agonist resveratrol 20mg/Kg at the same time, once a day, blood uric acid and creatinine concentration were detected at different time points through the canthus vein. The pathological changes of renal tissue were detected by HE staining.
Two, the effect of resveratrol on MSU induced acute arthritis in mice.
The model of gouty arthritis was induced by MSU in the foot of the 6-8 week old C57BL/6 male mice. The degree of joint swelling was evaluated by measuring the thickness of the foot of the foot by measuring the thickness of the palmar palmar. The degree of arthritis was evaluated by measuring the 99mTc absorption rate, and the different doses of resveratrol were given at different time points to evaluate the changes of arthritis.
Research results
The yeast ointment combined with potassium oxazine could establish a continuous and stable hyperuricemia model, in which the level of serum uric acid in the model 3-14d increased significantly and had no renal impairment, but the level of serum uric acid in the modeling 21d was slowed down, creatinine was slightly increased, the renal small balloon wall thickening, renal interstitial inflammatory cell infiltration and renal tubules were found in the kidney pathology. The epithelial cells were swollen. The intervention group was treated with resveratrol at the same time in the induced hyperuricic acid model. The serum uric acid level of the mice was significantly lower than that of the model group (P0.05), and the above renal pathological changes were improved.
Local injection of MSU can induce acute arthritis in mice. The degree of inflammation is proportional to the concentration of MSU, and it has a certain time limit. After MSU injection, the inflammation of the foot palms of 8-12h mice is the most obvious. After 24 hours, the inflammation gradually decreases. After the resveratrol pretreatment, the MSU induced arthritis is relieved (P0.05), resveratrol Resveratol was pretreated for 3 days, 7 days, and 14 days later, and then MSU induced inflammation was found to be pretreated for 14 days in mice with the lightest inflammation, and the effect was positively correlated with the dose of resveratrol.
research conclusion
1. yeast extract combined with potassium oxazinate can establish a sustained and stable hyperuricemia model.
2. Sirt1 agonist resveratrol can prevent and treat gout attacks through its lowering of uric acid and inflammatory regulation.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R589.7;R-332

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