左右归丸对EAE大鼠中枢神经系统CREBmRNA和蛋白表达的影响

Effects of Zuogui Pill and Yougui Pill on Expression of CREB mRNA and Protein in the Central Nervous System of Rats with EAE

目的:探讨实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)大鼠脑和脊髓中环磷酸腺苷反应元件结合蛋白(cAMP response element binding protein,CREB)mRNA和蛋白表达及左归丸和右归丸对其的调节作用。方法:将雌性Lewis大鼠随机分为正常组、模型组、激素组、左归丸组和右归丸组。给予正常组大鼠双后足垫注射生理盐水,其余各组注射髓鞘碱性蛋白(myelin basic protein,MBP)68-86抗原,免疫诱导EAE模型。分别灌胃给予相应的治疗药物或生理盐水。于14天(急性期)及28天(缓解期)分别取大鼠脑和脊髓,用实时荧光定量RT-PCR和Western-blot法检测CREB mRNA及蛋白的表达。结果:与正常组比较,EAE模型组大鼠脑和脊髓的CREB除了缓解期脑中蛋白的表达明显升高(P<0.05)外,其余均降低,特别是在急性期的大鼠脑中CREB mRNA及蛋白明显降低(P<0.05或P<0.01);急性期和缓解期脊髓中CREB mRNA也显著降低(P<0.01)。与模型组比较,左右归丸组急性期大鼠脊髓CREB mRNA和缓解期蛋白均升高明显(P<0.05或P<0.01)。左归丸组急性期大鼠脑CREB mRNA表达有显著上调(P<0.01),缓解期脑和急性期脊髓中的蛋白有明显上调(P<0.05或P<0.01)。结论:左右归丸对EAE大鼠的中枢神经损伤后的修复作用可能与上调CREB mRNA和蛋白表达有关。但二方在作用时间、部位和靶点各有侧重。

Objective： To study the expression of cAMP response element binding protein （CREB） mRNA and protein in the brain and spinal cord of rats with experimental autoimmune encephalomyelitis （EAE） and the regulative effect of Zuogui pill（ZGP） and Yougui pill（YGP）. Methods： Female Lewis rats were divided into normal control （NC）, EAE model （MO）, prednisone acetate, ZGP, and YGP -treated EAE groups. The pads of hind feet in NC rats were injected with normal saline （NS） and others were injected antigen containing myelin basic protein （MBP）68-86 to induce EAE models. The rats were treated by different medicines or NS. The brain and spinal cord were harvested on the 14th （acute period） and 28th（ remission period） day. The mRNA and protein of CREB in the brain and spinal cord of rats were detected by real - time quantitative poly-merase chain reaction and western blot method. Results： Compared with NC group, the protein and mRNA of CREB in the brain and spinal cord of rats in MO group were decreased except the brain protein in remission period were significantly increased （ P 〈 0.05 ）. Especially, the protein and mRNA of CREB in the brain in a- cute period were decreased significantly （ P 〈 0.05 or P 〈 0.01 ）, CREB mRNA of spinal cord in acute and remission period were also decreased significantly （P 〈 0.05）. Compared with MO group, the CREB mRNA in spinal cord in acute period and the protein in remission period of ZGP and YGP groups were increased sig- nificantly （ P 〈 0.05 or P 〈 0.01 ）. The brain CREB mRNA of ZGP group in acute period was increased sig- nificantly （P 〈 0.01 ） ; the proteins of brain in remission period and of spinal cord in acute period were in- creased significantly （P 〈0.05 or P 〈 0.01 ）. Conclusion： The repair effect of ZGP and YGP on EAE rats with damage of central nervous system may be related to the up - regulation of the protein and mRNA of CREB. But the effect of the two formulas was different in time, tissue and targets.