CAJ | 학술논문

目的观察慢性束缚应激对小鼠下丘脑腹内侧核(VMH)烟酰胺腺嘌呤二核苷酸磷酸盐(NADPH)氧化酶表达及活性氧(ROS)氧化应激损伤的影响。方法实验分为对照组和慢性束缚应激组,每周记录小鼠24 h摄食量和体重变化,用二氢乙啡啶(DHE)染色法检测 VMH ROS 的生成,HE 染色观察 VMH 形态学变化,免疫组化检测 VMH NOX2、p47phox 和 RAC1的表达,Western blot法检测下丘脑PKCα、NOX2、p47phox和RAC1的表达。结果与对照组比较,慢性束缚应激能明显降低小鼠24 h摄食量和体重；DHE染色结果显示,慢性束缚应激能明显增加小鼠VMH神经元ROS的生成；HE染色结果显示,慢性束缚应激能明显导致VMH神经元损伤；免疫组化和蛋白印迹结果显示,与对照组比较,慢性束缚应激小鼠VMH神经元PKCα、NOX2、p47phox和RAC1的表达明显增加。结论慢性束缚应激可导致小鼠VMH神经元损伤,其机制可能与增加VMH神经元NAD-PH氧化酶介导的ROS生成增多有关。
목적관찰만성속박응격대소서하구뇌복내측핵(VMH)연선알선표령이핵감산린산염(NADPH)양화매표체급활성양(ROS)양화응격손상적영향。방법실험분위대조조화만성속박응격조,매주기록소서24 h섭식량화체중변화,용이경을배정(DHE)염색법검측 VMH ROS 적생성,HE 염색관찰 VMH 형태학변화,면역조화검측 VMH NOX2、p47phox 화 RAC1적표체,Western blot법검측하구뇌PKCα、NOX2、p47phox화RAC1적표체。결과여대조조비교,만성속박응격능명현강저소서24 h섭식량화체중；DHE염색결과현시,만성속박응격능명현증가소서VMH신경원ROS적생성；HE염색결과현시,만성속박응격능명현도치VMH신경원손상；면역조화화단백인적결과현시,여대조조비교,만성속박응격소서VMH신경원PKCα、NOX2、p47phox화RAC1적표체명현증가。결론만성속박응격가도치소서VMH신경원손상,기궤제가능여증가VMH신경원NAD-PH양화매개도적ROS생성증다유관。
Objective To explore the effects of chronic restraint stress on NADPH oxidase (NOX) expression and the reactive oxygen species (ROS) generation in the hypothalamic ventromedial nucleus (VMH) in male mice. Methods Mice were divided into two groups:control group and chronic restraint stress group (4 weeks). And the body weight and food intake (24 h) were recorded every week. The ROS generation in VMH was detected by di-hydroethidium ( DHE). The histological changes were investigated by HE staining. The expression of NOX2, p47phox and RAC1 in VMH were examined by immunohistochemistry and immunoblot analysis. The expression of PKCα in VMH was measured by immunoblot analysis. Results Compared with control group, chronic restraint stress significantly decreased the food intake and body weight, and the ROS production was significantly increased in VMH in male mice. Histology results showed that chronic restraint stress significantly induce neuronal damage in VMH. Immunohistochemical and immunoblot analysis showed that, compared with control group, the expression of PKCα, NOX2, p47phox and RAC1 were significantly increased in VMH. Conclusion Chronic restraint stress may induce the VMH neuronal damage in male mice. The mechanisms may be related to NADPH oxidase mediated ROS accumulation and oxidative damage.