Abstract：
BACKGROUND: Research on the imbalance of proopiomelanocortin (POMC)/agouti-related protein (AgRP) neurons in the hypothalamus holds potential insights into the pathophysiology of diabetes. Jinkui Shenqi pills (JSP), a prevalent traditional Chinese medicine, regulate hypothalamic function and treat diabetes.
OBJECTIVE: To investigate the hypoglycemic effect of JSP and explore the probable mechanism in treating diabetes.
METHODS: A type 2 diabetes mouse model was used to investigate the pharmacodynamics of JSP. The glucose-lowering efficacy of JSP was assessed through various metrics including body weight, food consumption, fasting blood glucose (FBG), serum insulin levels, and an oral glucose tolerance test (OGTT). To elucidate the modulatory effects of JSP on hypothalamic mechanisms, we quantified the expression and activity of POMC and AgRP and assessed the insulin-mediated phosphoinositide 3-kinase (PI3K)/protein kinase A (AKT)/forkhead box O1 (FOXO1) pathway in diabetic mice via western blotting and immunohistochemistry. Additionally, primary hypothalamic neurons were exposed to high glucose and palmitic acid levels to induce insulin resistance, and the influence of JSP on POMC/AgRP protein expression and activation was evaluated by PI3K protein inhibition using western blotting and immunofluorescence.
RESULTS: Medium- and high-dose JSP treatment effectively inhibited appetite, resulting in a steady declining trend in body weight, FBG, and OGTT results in diabetic mice (p < 0.05). These JSP groups also had significantly increased insulin levels (p < 0.05). Importantly, the medium-dose group exhibited notable protection of hypothalamic neuronal and synaptic structures, leading to augmentation of dendritic length and branching (p < 0.05). Furthermore, low-, medium-, and high-dose JSP groups exhibited increased phosphorylated (p) INSR, PI3K, pPI3K, AKT, and pAKT expression, as well as decreased FOXO1 and increased pFOXO1 expression, indicating improved hypothalamic insulin resistance in diabetic mice (p < 0.05). Treatment with 10% JSP-enriched serum produced a marked elevation of both expression and activation of POMC (p < 0.05), with a concurrent reduction in AgRP expression and activation within primary hypothalamic neurons (p < 0.05). Intriguingly, these effects could be attributed to the regulatory dynamics of PI3K activity.
CONCLUSIONS: Our findings suggest that JSP can ameliorate diabetes by regulating POMC/AgRP expression and activity. The insulin-mediated PI3K/AKT/FOXO1 pathway plays an important regulatory role in this intricate process.
OBJECTIVE: To investigate the hypoglycemic effect of JSP and explore the probable mechanism in treating diabetes.
METHODS: A type 2 diabetes mouse model was used to investigate the pharmacodynamics of JSP. The glucose-lowering efficacy of JSP was assessed through various metrics including body weight, food consumption, fasting blood glucose (FBG), serum insulin levels, and an oral glucose tolerance test (OGTT). To elucidate the modulatory effects of JSP on hypothalamic mechanisms, we quantified the expression and activity of POMC and AgRP and assessed the insulin-mediated phosphoinositide 3-kinase (PI3K)/protein kinase A (AKT)/forkhead box O1 (FOXO1) pathway in diabetic mice via western blotting and immunohistochemistry. Additionally, primary hypothalamic neurons were exposed to high glucose and palmitic acid levels to induce insulin resistance, and the influence of JSP on POMC/AgRP protein expression and activation was evaluated by PI3K protein inhibition using western blotting and immunofluorescence.
RESULTS: Medium- and high-dose JSP treatment effectively inhibited appetite, resulting in a steady declining trend in body weight, FBG, and OGTT results in diabetic mice (p < 0.05). These JSP groups also had significantly increased insulin levels (p < 0.05). Importantly, the medium-dose group exhibited notable protection of hypothalamic neuronal and synaptic structures, leading to augmentation of dendritic length and branching (p < 0.05). Furthermore, low-, medium-, and high-dose JSP groups exhibited increased phosphorylated (p) INSR, PI3K, pPI3K, AKT, and pAKT expression, as well as decreased FOXO1 and increased pFOXO1 expression, indicating improved hypothalamic insulin resistance in diabetic mice (p < 0.05). Treatment with 10% JSP-enriched serum produced a marked elevation of both expression and activation of POMC (p < 0.05), with a concurrent reduction in AgRP expression and activation within primary hypothalamic neurons (p < 0.05). Intriguingly, these effects could be attributed to the regulatory dynamics of PI3K activity.
CONCLUSIONS: Our findings suggest that JSP can ameliorate diabetes by regulating POMC/AgRP expression and activity. The insulin-mediated PI3K/AKT/FOXO1 pathway plays an important regulatory role in this intricate process.
摘要：
背景：关于下丘脑中的前黑皮质素（POMC）/刺鼠相关蛋白（AgRP）神经元失衡的研究为糖尿病的病理生理学提供了潜在的见解。金葵肾气丸（JSP），一种流行的中药，调节下丘脑功能和治疗糖尿病。
目的：研究JSP的降血糖作用，探讨其治疗糖尿病的可能机制。
方法：采用2型糖尿病小鼠模型研究JSP的药效学。JSP的降糖功效通过各种指标进行评估，包括体重，食物消费,空腹血糖（FBG），血清胰岛素水平，和口服葡萄糖耐量试验（OGTT）。为了阐明JSP对下丘脑机制的调节作用，我们定量了POMC和AgRP的表达和活性,并通过蛋白质印迹和免疫组织化学评估了糖尿病小鼠中胰岛素介导的磷酸肌醇3-激酶(PI3K)/蛋白激酶A(AKT)/叉头盒O1(FOXO1)通路.此外,原发性下丘脑神经元暴露于高葡萄糖和棕榈酸水平诱导胰岛素抵抗，采用免疫印迹法和免疫荧光法通过PI3K蛋白抑制评价JSP对POMC/AgRP蛋白表达和活化的影响。
结果：中、大剂量JSP治疗可有效抑制食欲，导致体重稳步下降的趋势，FBG,和OGTT结果在糖尿病小鼠中(p<0.05)。这些JSP组的胰岛素水平也显著升高(p<0.05)。重要的是,中剂量组对下丘脑神经元和突触结构有明显的保护作用，导致树突长度和分支增加(p<0.05)。此外，低,medium-,高剂量JSP组显示磷酸化（p）INSR增加，PI3K,pPI3K,AKT,和pAKT表达式，以及减少FOXO1和增加pFOXO1表达，提示糖尿病小鼠下丘脑胰岛素抵抗改善(p<0.05)。用10%富含JSP的血清处理产生POMC表达和活化的显著升高(p<0.05),原发性下丘脑神经元内AgRP表达和激活同时减少（p<0.05）。有趣的是，这些效应可归因于PI3K活性的调节动力学.
结论：我们的研究结果表明，JSP可以通过调节POMC/AgRP的表达和活性来改善糖尿病。胰岛素介导的PI3K/AKT/FOXO1途径在这一复杂过程中起着重要的调节作用。
目的：研究JSP的降血糖作用，探讨其治疗糖尿病的可能机制。
方法：采用2型糖尿病小鼠模型研究JSP的药效学。JSP的降糖功效通过各种指标进行评估，包括体重，食物消费,空腹血糖（FBG），血清胰岛素水平，和口服葡萄糖耐量试验（OGTT）。为了阐明JSP对下丘脑机制的调节作用，我们定量了POMC和AgRP的表达和活性,并通过蛋白质印迹和免疫组织化学评估了糖尿病小鼠中胰岛素介导的磷酸肌醇3-激酶(PI3K)/蛋白激酶A(AKT)/叉头盒O1(FOXO1)通路.此外,原发性下丘脑神经元暴露于高葡萄糖和棕榈酸水平诱导胰岛素抵抗，采用免疫印迹法和免疫荧光法通过PI3K蛋白抑制评价JSP对POMC/AgRP蛋白表达和活化的影响。
结果：中、大剂量JSP治疗可有效抑制食欲，导致体重稳步下降的趋势，FBG,和OGTT结果在糖尿病小鼠中(p<0.05)。这些JSP组的胰岛素水平也显著升高(p<0.05)。重要的是,中剂量组对下丘脑神经元和突触结构有明显的保护作用，导致树突长度和分支增加(p<0.05)。此外，低,medium-,高剂量JSP组显示磷酸化（p）INSR增加，PI3K,pPI3K,AKT,和pAKT表达式，以及减少FOXO1和增加pFOXO1表达，提示糖尿病小鼠下丘脑胰岛素抵抗改善(p<0.05)。用10%富含JSP的血清处理产生POMC表达和活化的显著升高(p<0.05),原发性下丘脑神经元内AgRP表达和激活同时减少（p<0.05）。有趣的是，这些效应可归因于PI3K活性的调节动力学.
结论：我们的研究结果表明，JSP可以通过调节POMC/AgRP的表达和活性来改善糖尿病。胰岛素介导的PI3K/AKT/FOXO1途径在这一复杂过程中起着重要的调节作用。
