鲁斯可皂苷元对脑缺血再灌注损伤小鼠时钟基因的调控作用

Regulation of clock genes by ruscogenin in mice with cerebral ischemia/reperfusion injury

目的研究鲁斯可皂苷元(Rus)对小鼠脑缺血再灌注(I/R)损伤的改善作用及其机制。方法C57BL/6J小鼠分为假手术组、假手术+Rus组、I/R组和I/R+Rus组,采用大脑中动脉栓塞线栓法制备小鼠I/R模型,小鼠在术前1 h ig给予Rus 10 mg·kg^(-1)或同体积溶媒(0.5%羧甲纤维素钠)。再灌注24 h后,进行神经功能评分并测定脑梗死体积,采用实时荧光定量PCR和Western印迹法检测时钟基因Per1,Clock和Bmal1 mRNA和蛋白表达水平。结果与假手术组相比,I/R组神经功能评分增高(P<0.01),脑梗死体积增大(P<0.01),Per1,Clock和Bmal1 mRNA和蛋白表达水平下调(P<0.05,P<0.01);与I/R组相比,I/R+Rus组的脑梗死体积减小(P<0.01),神经功能评分降低(P<0.01),Per1,Clock和Bmal1 mRNA和蛋白表达水平上调(P<0.05,P<0.01)。结论 Rus可减轻小鼠脑I/R损伤,其机制可能与调控时钟基因的表达有关。

OBLECTIVE To investigate the regulatory effect of ruscogenin(Rus) on cortical clock genes in mice with cerebral ischemia/reperfusion(I/R) injury. METHODS I/R model was established by middle cerebral artery occlusion/reperfusion in C57 BL/6 J mice. The mice were randomly divided into four groups: sham, sham+Rus, I/R, and I/R+Rus. Mice were given Rus 10 mg·kg^(-1) or an equal volume of 0.5% sodium carboxymethyl cellulose by intragastric administration 1 h before middle cerebral artery occlusion. The percentage of right hemisphere brain infarct volume and neurological deficits score were evaluated after 24 h of reperfusion. Real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expressions of Per1, Clock and Bmal1. RESULTS Compared with the sham group, the percentage of right hemisphere brain infarct volume and neurological deficits score were increased in the I/R group(P<0.01), and the mRNA and protein expressions of Per1, Clock,Bmal1 were down-regulated in the I/R group(P<0.05, P<0.01). Compared with the I/R group, the cerebral infarct volume and neurological deficits score declined in I/R+Rus group(P<0.01), and the gene and protein expressions of Per1, Clock and Bmal1 were up-regulated in the I/R+Rus group(P<0.05, P<0.01). CONCLUSION Rus alleviates cerebral I/R injury and regulates the expressions of clock genes in mice. These effects are related to regulating the expressions of clock genes.