CIRP对冷应激小鼠血清生化指标、相关细胞免疫因子及能量代谢的影响

Effect of CIRP to biochemical indexs,related cellular immune factors and energy metabolism in serum of cold stress mice

将45只6周龄体质量(18±2)g SPF级雄性SD小鼠随机分为5组(n=5),常温对照组5只(A组);低温对照组冷刺激4、8h各5只(B组);冷诱导RNA结合蛋白(cold-inducible RNA binding protein,CIRP)过表达组冷刺激4、8h各5只(C组),注射CIRP过表达慢病毒液0.1mL;CIRP干扰组冷刺激4、8h各5只(D组),注射CIRP干扰病毒液0.1mL;阴性对照组冷刺激4、8h各5只(E组),注射空载体病毒液0.1mL。A组置于常温(21±0.1)℃条件下饲养,后4组置于低温(4±0.1)℃环境饲养,分别在4h及8h后通过摘除眼球的方法采取血液。采用ELISA法检测血清中IL-1、IL-2、IL-6、GSH-PX、SOD、T-AOC、MDA、丙酮酸、PFK-1、LDH的含量。结果显示:(1)冷应激处理后,小鼠血清中相关细胞因子水平出现明显变化,人为的致使CIRP在小鼠体内出现不同的表达对血清中相关细胞因子的影响差异显著(P<0.05);(2)冷应激处理后,小鼠血清中氧化还原指标GSH-PX、SOD、T-AOC、MDA变化非常明显,人为的致使CIRP在小鼠体内出现不同的表达对血清中相关氧化还原指标的影响差异极显著(P<0.01);(3)冷应激处理后,血清当中能量代谢相关指标无明显变化,人为的致使CIRP在小鼠体内出现不同的表达对血清中相关能量代谢指标的影响差异不显著(P>0.05)。结果表明:在不影响动物机体能量代谢的情况下,CIRP可以有效地抑制细胞内氧自由基的生成,从而提高小鼠抗氧化能力,以及调节动物机体的相关免疫因子,对小鼠抵抗低温应激起到积极地保护作用。

To explore the relationship between cold-inducible RNA-binding protein （CIRP） and bi- ochemical indicators, related cytokines and energy metabolism of anti- cold stress in mouse serum. Established mouse model of low temperature stress,45 SPF male SD mice were randomly divided into five groups（n= 5）.A normal temperature control group,B hypothermia group （for 4,8 h）℃ CIRP overexpression group （for 4,8 h） ,injected CIRP overexpression lentivirus fluid;D CIRP in- terference group（for 4,8 h） ,injected CIRP interfere lentivirus fluid;E negative control group （for 4,8 h）,injected empty vector lentivirus fluid. The mice of A group were reared under room tem- perature （21 ～ 0.1）℃, other group mice were reared under low environment （4±0.1） ℃. blood samples were taken by removal of the eye＇s method after 4 h and 8 h respectively,detected serum contents of IL-1, IL-2, IL-6, GSH-PX, SOD, T-AOC, MDA, pyruvate, PFK-1 and LDH content byELISA. The results showed that after treated with cold stress, （1） levels of cytokines related elis- played a significant change in serum,and was significantly different when treated with CIRP （P〈 0.05） ;（2） redox indicators,such as GSH-PX,SOD,T-AOC,MDA changed significantly in mouse serum （P〈0.01） ; （3） there was no significant difference in energy metabolism in serum （P〈0.05）. These results suggest that without prejudice to energy metabolism,CIRP could effectively suppress generation of intracellular oxygen free radicals,enhance the anti-oxidative ability of mice and regulate the animal＇s body related immune factors, therefore play an active protective role in resisting low temperature stress in mice.