左归降糖解郁方对模拟糖尿病并发抑郁症环境下海马神经元的影响
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摘要
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。
Objective To explore the effects of Zuogui Jiangtang Jieyu Prescription on hippocampal neurons in simulated diabetes mellitus with depression(DD); To discuss its possible mechanism. Methods Hippocampal neurons from fetal rats of 18 d pregnant rats were primitively cultivated, and the glucose combined with corticosterone was used to establish DD hippocampal neuronal cell model. Cultured hippocampal neurons were randomly divided into normal group, blank serum group, model group, positive drug(metformin plus fluoxetine) serum group, TCM(Zuogui Jiangtang Jieyu Prescription) serum group and blocker group. The normal group and the model group were given equal amount of culture, the blocker group was given 200 μmol/L Antalarmin, and the remaining groups were given 10% volume of drug serum or blank serum. After modeling intervention for 18 h, Nissl staining was used to detect hippocampal neurons injury. High content analysis(HCA) was used to detect the expressions of CRHR1, Cofilin and Actin protein, and real-time PCR was used to detect expressions of CRHR1, Cofilin and Actin mRNA. Results Compared with the normal group and blank serum group, neural network and dentritic spines ruptured or disappeared and the quantity of Nissl bodies decreased significantly, and expression of CRHR1 was significantly up-regulated(P<0.01), while the expressions of Cofilin and Actin were significantly down-regulated(P<0.05). Compared with the model group, positive drug serum group, TCM serum group and blocker group could partially reverse the above process(P<0.05, P<0.01). Conclusion Zuogui Jiangtang Jieyu Prescription has protective effects on hippocampal neurons in simulated DD environmen, and its mechanism is related to the regulation expressions of CRHR1, Cofilin and Actin.
Objective To explore the effects of Zuogui Jiangtang Jieyu Prescription on hippocampal neurons in simulated diabetes mellitus with depression(DD); To discuss its possible mechanism. Methods Hippocampal neurons from fetal rats of 18 d pregnant rats were primitively cultivated, and the glucose combined with corticosterone was used to establish DD hippocampal neuronal cell model. Cultured hippocampal neurons were randomly divided into normal group, blank serum group, model group, positive drug(metformin plus fluoxetine) serum group, TCM(Zuogui Jiangtang Jieyu Prescription) serum group and blocker group. The normal group and the model group were given equal amount of culture, the blocker group was given 200 μmol/L Antalarmin, and the remaining groups were given 10% volume of drug serum or blank serum. After modeling intervention for 18 h, Nissl staining was used to detect hippocampal neurons injury. High content analysis(HCA) was used to detect the expressions of CRHR1, Cofilin and Actin protein, and real-time PCR was used to detect expressions of CRHR1, Cofilin and Actin mRNA. Results Compared with the normal group and blank serum group, neural network and dentritic spines ruptured or disappeared and the quantity of Nissl bodies decreased significantly, and expression of CRHR1 was significantly up-regulated(P<0.01), while the expressions of Cofilin and Actin were significantly down-regulated(P<0.05). Compared with the model group, positive drug serum group, TCM serum group and blocker group could partially reverse the above process(P<0.05, P<0.01). Conclusion Zuogui Jiangtang Jieyu Prescription has protective effects on hippocampal neurons in simulated DD environmen, and its mechanism is related to the regulation expressions of CRHR1, Cofilin and Actin.
引文
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[6]凌佳,杨琴,刘检,等.左归降糖解郁方对模拟糖尿病并发抑郁症环境下模型大鼠海马神经元凋亡相关蛋白的影响[J].中国中医药信息杂志,2017,24(10):35-39.
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[8]孟盼,赵洪庆,杜青,等.左归降糖解郁方对糖尿病并发抑郁症大鼠海马神经元凋亡的影响[J].中国中医药信息杂志,2016,23(9):78-81.
[9]刘晔.突触可塑性及行为学相关的离子通道和信号传导通路及其他相关因素的研究[D].天津:南开大学,2014.
[10]ZHANG Y,ZHENG Y,XU Y,et al.Corticotropin-releasing hormone suppresses synapse formation in hippocampus of male rats via inhibition of CXCL5 secretion by glia[J].Endocrinology,2018,159(2):622-638.
[11]HOLSBOER F,ISING M.Central CRH system in depression and anxiety-evidence from clinical studies with CRH1 receptor antagonists[J].European Journal of Pharmacology,2008,583(2):350-357.
[12]LIAO X M,YANG X D,JIA J,et al.Blockade of corticotropinreleasing hormone receptor 1 attenuates early-life stressinduced synaptic abnormalities in the neonatal hippocampus[J].Hippocampus,2014,24(5):528-540.
[13]SHAW A E,BAMBURG J R.Peptide regulation of cofilin activity in the CNS:A novel therapeutic approach for treatment of multiple neurological disorders[J].Pharmacology&Therapeutics,2017,175:17-27.
[14]赵爽,邓佳卉,徐志卿,等.Cofilin表达调控与神经突触可塑性研究进展[J].神经解剖学杂志,2016,32(1):137-142.
[2]张苇,郑蔚,张丽,等.糖尿病并发症患者抑郁状况调查及影响因素研究[J].中国实用神经疾病杂志,2016,19(5):32-34.
[3]张娟,毕艳,沈山梅,等.2型糖尿病患者抑郁的患病率及相关危险因素分析[J].中华内分泌代谢杂志,2011,27(10):796-799.
[4]张秀丽,王宇红,杨蕙,等.左归降糖解郁方对糖尿病并发抑郁症模型大鼠学习记忆及海马超微结构的影响[J].时珍国医国药,2015,26(4):787-789.
[5]张秀丽,王宇红,杨蕙,等.左归降糖解郁方对大鼠海马神经元细胞的保护作用[J].湖南中医药大学学报,2014,34(12):8-12.
[6]凌佳,杨琴,刘检,等.左归降糖解郁方对模拟糖尿病并发抑郁症环境下模型大鼠海马神经元凋亡相关蛋白的影响[J].中国中医药信息杂志,2017,24(10):35-39.
[7]ZHENG Y,ZHANG Y M,NI X.Urocortin 2 but not urocortin 3 promotes the synaptic formation in hipppocampal neurons via induction of NGF production by astrocytes[J].Endocrinology,2016,157(3):1200-1210.
[8]孟盼,赵洪庆,杜青,等.左归降糖解郁方对糖尿病并发抑郁症大鼠海马神经元凋亡的影响[J].中国中医药信息杂志,2016,23(9):78-81.
[9]刘晔.突触可塑性及行为学相关的离子通道和信号传导通路及其他相关因素的研究[D].天津:南开大学,2014.
[10]ZHANG Y,ZHENG Y,XU Y,et al.Corticotropin-releasing hormone suppresses synapse formation in hippocampus of male rats via inhibition of CXCL5 secretion by glia[J].Endocrinology,2018,159(2):622-638.
[11]HOLSBOER F,ISING M.Central CRH system in depression and anxiety-evidence from clinical studies with CRH1 receptor antagonists[J].European Journal of Pharmacology,2008,583(2):350-357.
[12]LIAO X M,YANG X D,JIA J,et al.Blockade of corticotropinreleasing hormone receptor 1 attenuates early-life stressinduced synaptic abnormalities in the neonatal hippocampus[J].Hippocampus,2014,24(5):528-540.
[13]SHAW A E,BAMBURG J R.Peptide regulation of cofilin activity in the CNS:A novel therapeutic approach for treatment of multiple neurological disorders[J].Pharmacology&Therapeutics,2017,175:17-27.
[14]赵爽,邓佳卉,徐志卿,等.Cofilin表达调控与神经突触可塑性研究进展[J].神经解剖学杂志,2016,32(1):137-142.