Abstract:
This study aimed to determine the effect of age on CVLM C1 neuron glucoregulatory proteins in the feeding pathway. Male Sprague Dawley rats aged 3 months and 24 months old were divided into two subgroups: the treatment group with 2-deoxy-d-glucose (2DG) and the control group. Rat brains were dissected to obtain the CVLM region of the brainstem. Western blot was used to determine protein expression of tyrosine hydroxylase (TH), phosphorylated TH at Serine40 (pSer40TH), AMP-activated protein kinase (AMPK), phosphorylated AMPK (phospho AMPK), and neuropeptide Y Y5 receptors (NPY5R) in CVLM samples. Immunofluorescence was used to determine TH-, AMPK-, and NPY5R-like immunoreactivities among other brain coronal sections. Results obtained denote a decrease in basal TH phosphorylation levels and AMPK proteins and an increase in TH proteins among aged CVLM neurons. Increases in the basal immunoreactivity of TH+, AMPK+, NPY5R+, TH+/AMPK+, and TH+/NPY5R+ were also observed among old rats. Young treatment-group rats saw a decrease in TH phosphorylation and AMPK proteins following 2DG administration, while an increase in AMPK phosphorylation and a decrease in TH proteins were found among the old-treatment-group rats. These findings suggest the participation of CVLM C1 neurons in counter-regulatory responses among young and old rats. Altering protein changes in aged CVLM C1 neurons may attenuate responses to glucoprivation, thus explaining the decline in food intake among the elderly.
摘要:
本研究旨在确定年龄对摄食途径中CVLMC1神经元糖调节蛋白的影响。将3个月和24个月大的雄性SD大鼠分为两个亚组:2-脱氧-d-葡萄糖(2DG)治疗组和对照组。解剖大鼠的大脑以获得脑干的CVLM区域。Westernblot检测酪氨酸羟化酶(TH)蛋白表达,在Serine40磷酸化TH(pSer40TH),AMP激活蛋白激酶(AMPK),磷酸化AMPK(磷酸AMPK),CVLM样品中的神经肽YY5受体(NPY5R)。免疫荧光用于确定TH-,AMPK-,和其他脑冠状切片中的NPY5R样免疫反应性。获得的结果表明,在老年CVLM神经元中,基础TH磷酸化水平和AMPK蛋白降低,TH蛋白增加。TH+的基础免疫反应性增加,AMPK+,NPY5R+,TH+/AMPK+,在老年大鼠中也观察到TH/NPY5R。年轻的治疗组大鼠在2DG给药后,TH磷酸化和AMPK蛋白降低,而在老年治疗组大鼠中发现AMPK磷酸化增加和TH蛋白减少。这些发现表明CVLMC1神经元参与了年轻和老年大鼠的反调节反应。改变老年CVLMC1神经元的蛋白质变化可能会减弱对葡萄糖化的反应,从而解释了老年人食物摄入量的下降。