摘要
目的:研究Leptin在脑缺血性损伤神经元凋亡中的作用及其机制。方法:将75只雄性昆明小鼠完全随机分成3组,即假手术组、缺血/再灌注模型组、Leptin干预组;通过大脑中动脉栓塞(MCAO)复制小鼠局灶性脑缺血再灌注损伤模型,Leptin干预组在缺血0 min腹腔注射Leptin(1μg/g体重),TUNEL染色检测神经元凋亡,RT-PCR检测凋亡相关基因bcl-2和caspase-3 mRNA表达,免疫组化凋亡相关基因bcl-2和caspase-3蛋白水平的表达。结果:模型组脑缺血中心区神经元以坏死为主,与假手术组相比,其半影区神经元凋亡数量显著增多、促凋亡基因caspase-3和抑凋亡基因bcl-2的mRNA和蛋白表达水平均显著升高(P<0.01);与模型组比较,Leptin干预组半影区凋亡神经元数量显著减少、caspase-3 mRNA和蛋白表达水平显著降低(P<0.01),抑凋亡基因bcl-2 mRNA和蛋白表达水平显著升高(P<0.01)。结论:Leptin能够通过上调抑凋亡基因bcl-2表达,下调促凋亡基因caspase-3表达抑制神经元凋亡,在脑缺血性损伤中发挥神经保护作用。
Objective： To study the effect of leptin on neuron apoptosis in mice with cerebral ischemia injury. Methods： Seventy-five male Kuming mice were randomly divided into 3 groups： sham, model and leptin intervention group, respectively. Focal cerebral ischemia/ reperfusion injury model in mice was established by middle cerebral artery occlusion. Leptin intervention group was injected with leptin （ 1μg/ g weight, I.P. ） at 0 rain of ischemic injury. Neuron apeptosis was detected by TUNEL staining. The mRNA expression of apoptosis relative gene bcl-2 and caspase-3 were detected by RT-PCR. The protein expression of bcl-2 and caspase-3 were detected by immunohistochemistry. Results： In model group, most of the neurons in the central area of cerebral ischemia had necrosis obviously, and the amount of neuron apoptosis was much higher than that in sham group （ P 〈 0.01）. Compared with sham group, both expression of pro-apoptosis gene caspase-3 and anti-apoptosis gene bcl-2 increased significantly in model group （ P 〈 0.01 ）. Compared with model group, the amount of neuron apoptosis and expression level of caspase-3 were decreased significantly （ P 〈 0.01 ）, whereas the mRNA and protein expression of bcl-2 were increased sig- nificantly in leptin intervention group （P 〈 0.01 ）. Conclusion： Leptin could reduce neuron apoptosis through down-regulation the expression of caspase-3 and up-regulation the expression of bel-2. It suggests that leptin could play a neuroprotective role in cerebral ischemia injury.
出处
《中国应用生理学杂志》
CAS
CSCD
2016年第4期305-309,I0005,共6页
Chinese Journal of Applied Physiology
基金
国家自然科学基金资助项目(30670821)
关键词
瘦素
脑缺血
神经元
凋亡
神经保护
小鼠
leptin
cerebral ischemia
neuron
apoptosis
neuroprotection
mice
