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为了观察游泳运动后大鼠下丘脑室旁核(PVN)内c-fos mRNA及Fos免疫阳性神经元的表达变化,探讨下丘脑PVN对不同形式运动的调节机制,本实验将60只雄性SD大鼠随机分为对照组(n=10)和运动组(n=50),建立大鼠游泳运动模型(持续训练和间歇训练)。在运动结束后0、0.5、1、2、4h等不同时间点,冰水浴分离新鲜脑组织或用多聚甲醛灌注固定取脑,前者经RT-PCR反应,半定量分析运动后下丘脑PVN内c-fos mRNA的含量变化,后者经冰冻切片、免疫组织化学染色,观察Fos免疫阳性神经元的定位和分布情况。结果显示:(1)对照组大鼠下丘脑PVN内c-fos mRNA表达极低,而运动结束后c-fos mRNA表达明显升高。持续运动组逐渐升高,1h时达到最大值;间歇训练结束后2h,c-fos mRNA表达最强,然后回落,运动结束4h组c-fos mRNA仍较对照组显著增多(P<0.05);(2)运动大鼠Fos免疫阳性神经元的数目明显增多,特别是在PVN处。在室旁核小细胞部(pPVN),持续组游泳结束后1hFos阳性神经元的数目显著升高达峰值,而间歇组在运动结束后2h达峰值,同一时刻间歇组的表达高于持续运动组;室旁核大细胞部(mPVN)内,持续组Fos阳性神经元数在运动结束后持续升高,2h后显著下降,而间歇组各时刻阳性神经元的表达变化无统计学差异(P>0.05)。以上结果提示,下丘脑PVN在运动后机体调节中起重要作用,不同的运动方式可以产生不同的影响,且pPVN对不同形式运动性应激反应具有较高灵敏度。
To observe the changes of c-fos mRNA and Fos-immunoreactive neurons in paraventricular hypothalamic paraventricular nucleus (PVN) after swimming in rats, and to explore the regulation mechanism of hypothalamus PVN on different forms of exercise. In this experiment, 60 male SD rats The rats were randomly divided into control group (n = 10) and exercise group (n = 50), swimming rat model (continuous training and intermittent training) was established. At 0, 0.5, 1, 2, 4h and other time points after exercise, ice-water bath was used to separate fresh brain tissue or paraformaldehyde perfusion to fix the brain. The former was semi-quantitatively analyzed by RT-PCR after PVN c-fos mRNA content changes, the latter by frozen section, immunohistochemical staining Fos immunoreactive neurons localization and distribution. The results showed that: (1) The expression of c-fos mRNA in the hypothalamus PVN in the control group was extremely low, while the c-fos mRNA expression was significantly increased after the exercise. The expression of c-fos mRNA was the highest at 2h after the interval training, and then decreased. After 4h of exercise, the c-fos mRNA level was still significantly increased (P <0.05) compared with the control group. (2) The number of Fos immunoreactive neurons in motor rats increased significantly, especially at PVN. In the pPVN group, the number of Fos positive neurons significantly increased at 1 hour after the end of continuous swimming group, while the peak value reached the peak at 2 hours after intermittent exercise in intermittent group, and the expression of intermittent group was higher than that of continuous exercise group at the same time. In mPVN group, the number of Fos positive neurons in persistent group continued to increase after exercise and decreased significantly after 2 h, but there was no significant difference in the expression of positive neurons in intermittent group (P> 0.05) ). These results suggest that the hypothalamus PVN play an important role in the regulation of the body after exercise, different exercise modes can have different effects, and pPVN has higher sensitivity to different forms of exercise stress response.