糖尿病大鼠下丘脑Nesfatin-1对胃运动的影响及机制研究

The Effect of Nesfatin-1 in Gastric Motility in Diabetic Rat

目的:探讨下丘脑腹内侧核Nesfatin-1对正常大鼠及糖尿病大鼠胃运动的影响及其潜在机制。方法:正常大鼠随机分为0.08μg,0.8μg,8.0μg/0.5μL Nesfatin-1组;30μg/0.5μL astressin-B组;(0.8μg Nesfatin-1+30μg astressin-B)/0.5μL组;0.5μL生理盐水(NS)组;正常羊血清+假刺激(NR+SS)组;正常羊血清+电刺激(NR+ES)组;抗NUCB2/Nesfatin-1抗体+假刺激(anti-Nn-Ab+SS)组;抗NUCB2/Nesfatin-1抗体+电刺激(anti-Nn-Ab+ES)组。制作糖尿病大鼠模型,将糖尿病大鼠随机分为0.08μg/0.5μL Nesfatin-1组;0.8μg/0.5μLNesfatin-1组;8.0μg/0.5μL Nesfatin-1组;0.5μLNS组;NR+SS组;NR+ES组;anti-Nn-Ab+SS组;anti-Nn-Ab+ES组。大鼠胃部置入感应器后腹内侧核置管,记录清醒大鼠胃运动及电刺激海马CA1区后的胃运动。结果:与生理盐水组相比,下丘脑腹内侧核注射不同浓度Nesfatin-1,大鼠胃收缩幅度和频率显著降低,下丘脑腹内侧核注射0.5μL(0.8μg Nesfatin-1+30μg astressin-B)混合液后,相比单独给予0.8μg Nesfatin-1组,大鼠胃收缩幅度和频率显著升高。大鼠下丘脑腹内侧核注射0.5μL Nesfatin-1(0.8μg),大鼠胃收缩幅度和频率显著降低,下丘脑腹内侧核注射0.5μL(0.8μg Nesfatin-1+30μg astressin-B)混合液后,相比单独给予0.8μg Nesfatin-1组,大鼠胃收缩幅度和频率显著升高。下丘脑腹内侧核注射抗NUCB2/Nesfatin-1抗体后再电刺激海马CA1区,与正常羊血清+电刺激组相比,大鼠胃收缩幅度和频率进一步增强,下丘脑腹内侧核注射抗NUCB2/Nesfatin-1抗体后再电刺激海马CA1区,与单独注射抗NUCB2/Nesfatin-1抗体+假电刺激组相比,大鼠的胃收缩幅度和频率显著增高。下丘脑腹内侧核注射抗NUCB2/Nesfatin-1抗体后再给予电刺激海马CA1区,与正常羊血清+电刺激组相比,正常大鼠和糖尿病大鼠胃运动指数均显著增加,下丘脑腹内侧核注射抗NUCB2/Nesfatin-1抗体后再电刺激海马CA1区,与单独注射抗NUCB2/Nesfatin-1抗体+假电刺激组相比,正常和糖尿病大鼠的胃运动指数均显著增高。与正常大鼠相比,电刺激海马CA1区、下丘脑腹内侧核注射抗NUCB2/Nesfatin-1抗体后再给予电刺激海马CA1区,或下丘脑腹内侧核微量注射抗NUCB2/Nesfatin-1抗体,糖尿病大鼠胃运动指数均无显著差异。结论:海马-下丘脑Nesfatin-1信号通路参与胃传入信息和胃运动调控,该作用可能与CRF系统活动有关。

Objective： To observe the effect of Nesfatin-1 on the gastric motility of normal rat and diabetic rats and its potential mechanism. Methods： Normal rats were randomly divided into 0.08 μg, 0.8 μg, 8.0 μg/0.5 μL Nesfatin-1 group; 30 μg/0.5 μL astressin-B group; （0.8 μg Nesfatin-1 ＋ 30 ixgastressin-B） / 0.5 μL group; （NR ＋ SS） group; normal sheep serum ＋ stimulated （NR ＋ ES） group; anti-NUCB2 / Nesfatin-1 antibody ＋ anti-Nn-Ab ＋ SS） group; anti-NUCB2/Nesfatin-1 antibody ＋ anti-Nn-Ab ＋ ES group. The diabetic rats were randomly divided into 0.08 ixg/0.5 IμL Nesfatin-1 group, 0.8 ixg/0.5 μL Nesfatin-1 group, 8.0 μg/0.5 μL Nesfatin-1 group, 0.5 μL NS group, NR ＋ SS NR ＋ ES group; anti-Nn-Ab ＋ SS group; anti-Nn-Ab ＋ ES group. After the rat stomach was placed in the sensor, put a catheter into ventral medial nucleus, then observe the gastric motility of conscious rats and after electrical stimulation of hippocampal CA1 area. Results： Compared with the normal saline group, Nesfatin-1 was injected into the ventral nucleus of the hypothalamus, and the amplitude and frequency of gastric contraction were significantly decreased. 0.5 μL （0.8 μg Nesfatin-1 ＋ 30 μg astressin-B） after mixed solution, the amplitude and frequency of gastric contraction were significantly increased in rats compared with 0.8 μg Nesfatin-1 alone. Rats were injected with 0.5 μL of Nesfatin-1 （0.8 μg） in the ventral medial nucleus of the hypothalamus, and the amplitude and frequency of gastric contraction were significantly decreased in the ventral medial nucleus of the hypothalamus. 0.5 μL （0.8 μg Nesfatin-1 ＋ 30 μg astressin-B） after treatment, the amplitude and frequency of gastric contraction were significantly higher in rats than 0.8 μg Nesfatin-1 alone. In the hypothalamic nucleus of the hypothalamus, the anti-NUCB2/Nesfatin-1 antibody was used to stimulate the hippocampal CA1 region, and the amplitude and frequency of gastric contraction were further enhanced compared with the normal sheep serum ＋/Nesfatin-1 antibody was used to stimulate the hippocampal CA1 region. Compared with the anti-NUCB2/Nesfatin-1 antibody ＋ pseudo-electrical stimulation group, the amplitude and frequency of gastric contraction were significantly increased. The NAGB2/Nesfatin-1 antibody was injected into the ventral medial nucleus of the hypothalamus to induce the electrical stimulation of the hippocampal CA1 region. Compared with the normal sheep serum ＋ electric stimulation group, the gastric motility index of the normal rats and diabetic rats were significantly increased, The gastric motility index of normal and diabetic rats was significantly higher than that of anti-NUCB2/Nesfatin-1 antibody ＋ pseudo-electrical stimulation group after injection of anti-NUCB2/Nesfatin-1 antibody in the hippocampal CA1 region. Compared with normal rats, the rats were injected with anti-NUCB2/Nesfatin-1 antibody in the hippocampal CA1 region and the hypothalamic ventral nucleus. The rats were injected with anti-NUCB2 / Nesfatin-1 antibody. There was no significant difference in gastric motility index between diabetic rats. Conclusion： Nesfatin-1 signaling pathway is involved in gastric afferent information and gastric motility regulation, which may be related to CRF system activity.