目的观察左归降糖解郁方对模拟糖尿病并发抑郁症(DD)环境下海马神经元的影响,探讨其可能的作用机制。方法海马神经元原代细胞取自受孕18 d SD大鼠胎鼠,葡萄糖联合皮质酮干预构建DD海马神经元细胞模型。将培养的海马神经元随机分为正常...目的观察左归降糖解郁方对模拟糖尿病并发抑郁症(DD)环境下海马神经元的影响,探讨其可能的作用机制。方法海马神经元原代细胞取自受孕18 d SD大鼠胎鼠,葡萄糖联合皮质酮干预构建DD海马神经元细胞模型。将培养的海马神经元随机分为正常组、空白血清组、模型组、阳性药(二甲双胍+氟西汀)药物血清组、中药(左归降糖解郁方)药物血清组和阻断剂组。正常组和模型组给予等量培养液,阻断剂组给予促肾上腺皮质激素释放激素1型受体(CRHR1)阻断剂安塔拉明200μmol/L,其余组给予体积分数为10%的药物血清或空白血清,造模干预18 h后,尼氏染色检测海马神经元损伤情况,高内涵细胞成像分析(HCA)技术检测CRHR1、突触可塑性相关蛋白切丝蛋白(Cofilin)和肌动蛋白(Actin)蛋白的表达,实时荧光定量PCR检测CRHR1、Cofilin和Actin mRNA的表达。结果与正常组及空白血清组比较,模型组海马神经元神经网络、树突棘断裂或消失,尼氏小体数量显著减少,CRHR1表达明显上调(P<0.01),Cofilin和Actin表达明显下调(P<0.05);与模型组比较,阳性药药物血清组、中药药物血清组和阻断剂组可一定程度上逆转上述变化(P<0.05,P<0.01)。结论左归降糖解郁方对模拟DD环境下海马神经元具有保护作用,其机制与调控CRHR1、Cofilin、Actin的表达有关。展开更多
OBJECTIVE:To study the development mechanism of kidney-Yang deficiency through the establishment of support vector machine models of relevant hormones of the pituitary-target gland axes in rats with kidney-Yang defici...OBJECTIVE:To study the development mechanism of kidney-Yang deficiency through the establishment of support vector machine models of relevant hormones of the pituitary-target gland axes in rats with kidney-Yang deficiency syndrome.METHODS:The kidney-Yang deficiency rat model was created by intramuscular injection of hydrocortisone,and contents of the hormones of the pituita- ry-thyroid axis:thyroid stimulating hormone(TSH),3,3',5-triiodothyronine(T_3) and thyroxine(T_4);hormones of the pituitary-adrenal gland axis:adrenocorticotropic hormone(ACTH) and Cortisol(CORT);and hormones of the pituitary-gonadal axis:luteinizing hormone(LH),follicle-stimulating hormone(FSH),and testosterone(T),were determined in the early,middle,and advanced stages.Ten support vector regression(SVR) models of the hormones were established to analyze the mutual relationships among the hormones of the three axes.RESULTS:The feedback control action of the pituitary-adrenal axis began to lose efficacy from the middle stage of kidney-Kong deficiency.The contents all hormones of the three pituitary-target gland axes decreased in the advanced stage.Relative errors of the jackknife test of the SVR models all were less than 10%.CONCLUSION:Imbalances in mutual regulation among the hormones of the pituitary-target gland axes,especially loss of effectiveness of the pituitary-adrenal axis,is one pathogenesis of kidney-Yang deficiency.The SVR model can accurately reflect the complicated non-linear relationships among pituitary-target gland axes in rats with of kidney-Yang deficiency.展开更多
文摘目的观察左归降糖解郁方对模拟糖尿病并发抑郁症(DD)环境下海马神经元的影响,探讨其可能的作用机制。方法海马神经元原代细胞取自受孕18 d SD大鼠胎鼠,葡萄糖联合皮质酮干预构建DD海马神经元细胞模型。将培养的海马神经元随机分为正常组、空白血清组、模型组、阳性药(二甲双胍+氟西汀)药物血清组、中药(左归降糖解郁方)药物血清组和阻断剂组。正常组和模型组给予等量培养液,阻断剂组给予促肾上腺皮质激素释放激素1型受体(CRHR1)阻断剂安塔拉明200μmol/L,其余组给予体积分数为10%的药物血清或空白血清,造模干预18 h后,尼氏染色检测海马神经元损伤情况,高内涵细胞成像分析(HCA)技术检测CRHR1、突触可塑性相关蛋白切丝蛋白(Cofilin)和肌动蛋白(Actin)蛋白的表达,实时荧光定量PCR检测CRHR1、Cofilin和Actin mRNA的表达。结果与正常组及空白血清组比较,模型组海马神经元神经网络、树突棘断裂或消失,尼氏小体数量显著减少,CRHR1表达明显上调(P<0.01),Cofilin和Actin表达明显下调(P<0.05);与模型组比较,阳性药药物血清组、中药药物血清组和阻断剂组可一定程度上逆转上述变化(P<0.05,P<0.01)。结论左归降糖解郁方对模拟DD环境下海马神经元具有保护作用,其机制与调控CRHR1、Cofilin、Actin的表达有关。
基金Supported by National Natural Science Foundation for Young Scholars of China(Study on the Mechanism of Kidney-Yang Deficiency Regulation with Yougui Pill Base on Support Vector Regression Machine,No.81403153)National Natural Science Foundation of China(General Program,Study on the Mechanisms of the Premature Ovarian Failure Regulation by Herbs Couples of Cuscuta Chinessis-Radix Bupleuri in Dingjing Decoction based on RBF Artificial Neural Networks,No.81073073) and National Natural Science Foundation of China(General Program,Study on the law of compatibility of Categorized Formula for Tonifying Kidney Yang based on Rough Set,No.30973977)
文摘OBJECTIVE:To study the development mechanism of kidney-Yang deficiency through the establishment of support vector machine models of relevant hormones of the pituitary-target gland axes in rats with kidney-Yang deficiency syndrome.METHODS:The kidney-Yang deficiency rat model was created by intramuscular injection of hydrocortisone,and contents of the hormones of the pituita- ry-thyroid axis:thyroid stimulating hormone(TSH),3,3',5-triiodothyronine(T_3) and thyroxine(T_4);hormones of the pituitary-adrenal gland axis:adrenocorticotropic hormone(ACTH) and Cortisol(CORT);and hormones of the pituitary-gonadal axis:luteinizing hormone(LH),follicle-stimulating hormone(FSH),and testosterone(T),were determined in the early,middle,and advanced stages.Ten support vector regression(SVR) models of the hormones were established to analyze the mutual relationships among the hormones of the three axes.RESULTS:The feedback control action of the pituitary-adrenal axis began to lose efficacy from the middle stage of kidney-Kong deficiency.The contents all hormones of the three pituitary-target gland axes decreased in the advanced stage.Relative errors of the jackknife test of the SVR models all were less than 10%.CONCLUSION:Imbalances in mutual regulation among the hormones of the pituitary-target gland axes,especially loss of effectiveness of the pituitary-adrenal axis,is one pathogenesis of kidney-Yang deficiency.The SVR model can accurately reflect the complicated non-linear relationships among pituitary-target gland axes in rats with of kidney-Yang deficiency.
