低氧诱导大鼠内脏脂肪组织炎症状态对SePP1的影响

The Affect to SePP1 of Hypoxia-induced Inflammatory State of Visceral Adipose Tissue

目的:既往研究显示SePP1具有一定的抗氧化作用,而随着年龄的增加机体逐步出现一个慢性低氧、炎症状态,我们通过4%O2浓度体外培养大鼠脂肪前体细胞模拟其体内的低氧状态,进而观察常氧(21%O2)及低氧(4%O2)状态下大鼠脂肪前体细胞中炎症因子(IL-6,MCP-1,SePP1)水平的变化及不同状态下硒蛋白SePP1水平的变化。方法:取6-8周SD大鼠肾周脂肪前体细胞,分别于常氧(21%O2)及低氧(4%O2)状态下进行体外培养,诱导分化后采用油红O染色进行鉴定,至第三代后,分别采用PCR及Western Blot技术检测两种状态下脂肪前体细胞中IL-6,MCP-1,SePP1基因及蛋白表达的不同变化,同时观察不同氧浓度对脂肪前体细胞增殖的影响。结果:4%氧浓度状态下培养的脂肪前体细胞中IL-6,MCP-1的基因及蛋白表达均明显高于正常氧浓度下的脂肪前体细胞,而SePP1的基因及蛋白表达均下降,且低氧状态下脂肪前体细胞增殖较常氧状态下加快。结论:低氧培养可进一步使机体内脏脂肪组织堆积加重,造成脂肪前体细胞的炎症状态,并且可导致SePP1的表达下降,而SePP1具有一定的抗氧化作用,与机体动脉粥样硬化等心血管疾病的发生、发展有一定的关联,本实验结论为通过干预体内SePP1的水平为靶点治疗动脉粥样硬化提供了一定的实验依据,为进一步研究SePP1在低氧状态下对动脉粥样硬化的影响及作用机制提供了一定的试验基础。

Objective： Previous studies have shown that SePP1 has antioxidant effects. With the increase of the body age the chronic hypoxia, the inflammation state are increasing, to observe the changes of inflammatory status,we cultured rat preadipocytes under 4%02 concentration in vitro （IL-6, MCP-1, SePP1 ）and different states selenoproteins SePP1 in rat preadipocyte in the state of normoxic （21% 02） and hypoxia （4% 02）. Methods： Preadipocytes from 6-8 weeks SD rats were cultured in vitro under normoxic and hypoxic state, red oil O staining identified after differentiate induced, for the third generation, the gene and protein expression in IL-6, MCP-1, SePP 1 were observed through the PCR and Western Blot technology under the two states, while observing the effects of different oxygen concentration on the proliferation of preadipocytes. Results： In preadipocytes cultured in 4% oxygen concentration state, IL-6, MCP-1 gene and protein expression were significantly higher than under normal oxygen concentration, but SePP1 gene and protein expression were decreased, also, preadipocytes proliferation in hypoxia compared with normoxic state were accelerated. Conclusions： Hypoxic state can further accumulation of visceral adipose tissue in the body heavier, increase preadipocyte inflammatory state decline and cause SePP1 expression. While SePP1 has some antioxygenation, is related with the occurrence and development of cardiovascular disease including atherosclerosis. The conclusions of this experiment provide a new basis for the treatment of arteriosclerosis though intervention SePP1 level in vivo, also provide the experimental basis for further study of the impact of SePP1 level under hypoxia with atherosclerosis and the mechanism between them.