The clinical effect of electroacupuncture on depression is widely recognized. However, the signal transduction pathways and target proteins involved remain unclear. In the present study, rat models of chronic restrain...The clinical effect of electroacupuncture on depression is widely recognized. However, the signal transduction pathways and target proteins involved remain unclear. In the present study, rat models of chronic restraint stress were used to explore the mechanism by which electroacupuncture alleviates depression. Rats were randomly divided into control, model, and electroacupuncture groups. Chronic restraint stress was induced in the model and electroacupuncture groups by restraining rats for 28 days. In the electroacupuncture group, electroacupuncture pretreatment at Baihui（GV20） and Yintang（GV29） acupoints was performed daily（1 m A, 2 Hz, discontinuous wave, 20 minutes） prior to restraint for 28 days. Open field tests and body weight measurements were carried out to evaluate the depressive symptoms at specific time points. On day 28, the crossing number, rearing number, and body weights of the model group were significantly lower than those in the control group. Behavior test results indicated that rat models of depressive-like symptoms were successfully established by chronic restraint stress combined with solitary raising. On day 28, an isobaric tag for a relative and absolute quantitation-based quantitative proteomic approach was performed to identify differentially expressed proteins in hippocampal samples obtained from the model and electroacupuncture groups. The potential function of these differential proteins was predicted through the use of the Cluster of Orthologous Groups of proteins（COG） database. Twenty-seven differential proteins（uncharacteristic proteins expected） were selected from the model and electroacupuncture groups. In addition to unknown protein functions, COG are mainly concentrated in general prediction function, mechanism of signal transduction, amino acid transport and metabolism groups. This suggests that electroacupuncture improved depressive-like symptoms by regulating differential proteins, and most of these related proteins exist in nerve cells.展开更多
Objective： To analyze the proteomic characteristics of Gan （肝））--ssttaaggnnaannccyy syndrome （（GGSSSS）） by seeking the differential protein in blood and tissues of GSS model rats.Methods： GSS model rats were...Objective： To analyze the proteomic characteristics of Gan （肝））--ssttaaggnnaannccyy syndrome （（GGSSSS）） by seeking the differential protein in blood and tissues of GSS model rats.Methods： GSS model rats were established by chronic restraint stress,keeping rats in restrain chamber for 6 h every day for 21 successive days.Their blood and liver samples were collected at the end of experiment for differential protein detection with methods of isoelectrofocusing and polyacrylamide SDS-PAGE,silver staining,and scanning.The gel images were analyzed with Imagemaster 2D Elite software,and the excavated differential protein spots were identified with matrix assistant laser resolving TOF mass spectrometry,Western blot,ELISA,and RT-PCR,respectively.Results： A method for isolating the protein in blood serum and tissues by two-dimensional gel electrophoresis was established and optimized.Six serum proteins and three liver proteins that differentially expressed were identified.The down-regulated differential proteins in serum of GSS model rats were serum albumin precursor,beta 1 globin,antibody against muscle acetylcholine receptor,Ig lambda-2 C region,and transthyretin （TTR）,and those in liver tissue were aryl sulfotransferase,enoyl-CoA hydratase,and TTR.TTR down-regulation was found in both serum and liver.Preliminary biological information analysis showed that these differential proteins involved in immune,neuroendocrine,nutrition,and substance metabolism.Conclusion： Proteomic analysis of differential proteins showed that TTR,aryl sulfotransferase,and enoyl-CoA hydratase expressions are downregulated in the GSS model rats,suggesting that the susceptibility of cancer could be enhanced by chronic stress.展开更多
The depressant-like effects of albiflorin(AF) were studied on stressed chronic restraint stress(CRS) rats. Experimental rats were subjected to immobilization stress for a daily 6 h-restraining in a plastic restrainer ...The depressant-like effects of albiflorin(AF) were studied on stressed chronic restraint stress(CRS) rats. Experimental rats were subjected to immobilization stress for a daily 6 h-restraining in a plastic restrainer for continuous 21 d and were treated with 30 or 15 mg·kg-1 of AF for 21 d. Control rats were maintained in completely non stressed conditions. Behavioral tests and biochemical analysis were applied to investigating a regulatory mechanism of anti-stress of AF. Treatment with AF significantly restored the depressant-like behaviors. Besides, AF increased the levels of 5-hydroxytryptophan(5-HT), 5-hydroxyindoleacetic acid(5-HIAA),noradrenaline(NE) and dopamine(DA) in the hippocampus and increased the level of brain-derived neurotrophic factor(BDNF) in serum and protein expression in hippocampus. In addition, AF decreased the levels of hypothalamo-pituitary-adrenal(HPA) cascade,reduced the level of NO and c GMP in serum and inhibited the overexpression of 5-HT2AR m RNA and protein expression. Taken together, AF can modulate the NO-mediated network pathway in the hippocampus against stress-induced depressive-like behaviors.These physiological and behavioral changes allow rats to avoid potential deleterious effects of stress that may result from chronically elevated levels of glucocorticosteroids over days.展开更多
Objective:To explore the effects of CUMS combined with CRS on mouse hippocampal glial cells and synaptic plasticity-related proteins. Methods: Forty mice were randomly divided into normal group (n=20) and model group ...Objective:To explore the effects of CUMS combined with CRS on mouse hippocampal glial cells and synaptic plasticity-related proteins. Methods: Forty mice were randomly divided into normal group (n=20) and model group (n=20). The model group used CUMS combined with CRS to prepare a mouse model of depression for 7 weeks. The behavioral evaluation of the mice at 3 weeks and 7 weeks after modeling was performed by sugar water preference test, open field test and tail suspension test. After the experiment, the samples were collected, and the content of TNF-a in the hippocampus of mice was detected by enzyme-linked immunosorbent assay. Immunohistochemical method was used to detect the Iba-1 and GFAP MOD values of mouse hippocampal CA1 area, CA3 area and DG area. Western blot was used to detect the protein expression of Iba-1, GFAP, SYN1 and PSD-95 in the hippocampus. fluorescence quantitative PCR method was used to detect the expression of SYN1, PSD-95 mRNA in hippocampus. Results: At the 3rd week after modeling, the body weight, sugar water preference rate, total distance moved, number of standing uprights, and stay time in the central area of the mice in the model group were all lower than those in the normal group (P<0.05), and the tail suspension immobility time was longer than that in the normal group (P<0.01). After 7 weeks of modeling, the body weight, sugar water preference rate, total distance moved, number of erection times, central area residence time, and average movement speed of the mice in the model group were lower than those in the normal group (P< 0.05), the tail suspension immobility time was longer than that in the normal group (P<0.01). The contents of TNF-a in the hippocampus were higher than those in the normal group (P<0.05). The GFAP MOD value and the relative expression of GFAP protein in hippocampal CA1, CA3 and DG regions were significantly lower than those in the normal group (P<0.05). The Iba-1 MOD value and the relative expression of Iba-1 protein in hippocampal CA1, CA3 and DG regions were significantly higher than those in the normal group (P<0.05). The relative expression of SYN1 and PSD-95 protein and the relative expression of SYN1 and PSD-95 mRNA in the hippocampus were significantly lower than those in the normal group (P<0.05). Conclusion: After 3 weeks of CUMS and CRS modeling, the depression-like behavior of mice appeared, and the depression of mice was more obvious after 7 weeks of modeling. The depression mouse model made by CUMS combined with CRS method may be related to increased hippocampal inflammation, excessive activation of microglia, decreased number of astrocytes and decreased synaptic plasticity.展开更多
基金supported by the National Natural Science Foundation of China,No.81373729
文摘The clinical effect of electroacupuncture on depression is widely recognized. However, the signal transduction pathways and target proteins involved remain unclear. In the present study, rat models of chronic restraint stress were used to explore the mechanism by which electroacupuncture alleviates depression. Rats were randomly divided into control, model, and electroacupuncture groups. Chronic restraint stress was induced in the model and electroacupuncture groups by restraining rats for 28 days. In the electroacupuncture group, electroacupuncture pretreatment at Baihui（GV20） and Yintang（GV29） acupoints was performed daily（1 m A, 2 Hz, discontinuous wave, 20 minutes） prior to restraint for 28 days. Open field tests and body weight measurements were carried out to evaluate the depressive symptoms at specific time points. On day 28, the crossing number, rearing number, and body weights of the model group were significantly lower than those in the control group. Behavior test results indicated that rat models of depressive-like symptoms were successfully established by chronic restraint stress combined with solitary raising. On day 28, an isobaric tag for a relative and absolute quantitation-based quantitative proteomic approach was performed to identify differentially expressed proteins in hippocampal samples obtained from the model and electroacupuncture groups. The potential function of these differential proteins was predicted through the use of the Cluster of Orthologous Groups of proteins（COG） database. Twenty-seven differential proteins（uncharacteristic proteins expected） were selected from the model and electroacupuncture groups. In addition to unknown protein functions, COG are mainly concentrated in general prediction function, mechanism of signal transduction, amino acid transport and metabolism groups. This suggests that electroacupuncture improved depressive-like symptoms by regulating differential proteins, and most of these related proteins exist in nerve cells.
基金Supported by the National Natural Science Foundation of China(No.30371713,30672769)
文摘Objective： To analyze the proteomic characteristics of Gan （肝））--ssttaaggnnaannccyy syndrome （（GGSSSS）） by seeking the differential protein in blood and tissues of GSS model rats.Methods： GSS model rats were established by chronic restraint stress,keeping rats in restrain chamber for 6 h every day for 21 successive days.Their blood and liver samples were collected at the end of experiment for differential protein detection with methods of isoelectrofocusing and polyacrylamide SDS-PAGE,silver staining,and scanning.The gel images were analyzed with Imagemaster 2D Elite software,and the excavated differential protein spots were identified with matrix assistant laser resolving TOF mass spectrometry,Western blot,ELISA,and RT-PCR,respectively.Results： A method for isolating the protein in blood serum and tissues by two-dimensional gel electrophoresis was established and optimized.Six serum proteins and three liver proteins that differentially expressed were identified.The down-regulated differential proteins in serum of GSS model rats were serum albumin precursor,beta 1 globin,antibody against muscle acetylcholine receptor,Ig lambda-2 C region,and transthyretin （TTR）,and those in liver tissue were aryl sulfotransferase,enoyl-CoA hydratase,and TTR.TTR down-regulation was found in both serum and liver.Preliminary biological information analysis showed that these differential proteins involved in immune,neuroendocrine,nutrition,and substance metabolism.Conclusion： Proteomic analysis of differential proteins showed that TTR,aryl sulfotransferase,and enoyl-CoA hydratase expressions are downregulated in the GSS model rats,suggesting that the susceptibility of cancer could be enhanced by chronic stress.
基金supported by the National Nature Science Foundation of China (Nos. 81473370 and 81173569)。
文摘The depressant-like effects of albiflorin(AF) were studied on stressed chronic restraint stress(CRS) rats. Experimental rats were subjected to immobilization stress for a daily 6 h-restraining in a plastic restrainer for continuous 21 d and were treated with 30 or 15 mg·kg-1 of AF for 21 d. Control rats were maintained in completely non stressed conditions. Behavioral tests and biochemical analysis were applied to investigating a regulatory mechanism of anti-stress of AF. Treatment with AF significantly restored the depressant-like behaviors. Besides, AF increased the levels of 5-hydroxytryptophan(5-HT), 5-hydroxyindoleacetic acid(5-HIAA),noradrenaline(NE) and dopamine(DA) in the hippocampus and increased the level of brain-derived neurotrophic factor(BDNF) in serum and protein expression in hippocampus. In addition, AF decreased the levels of hypothalamo-pituitary-adrenal(HPA) cascade,reduced the level of NO and c GMP in serum and inhibited the overexpression of 5-HT2AR m RNA and protein expression. Taken together, AF can modulate the NO-mediated network pathway in the hippocampus against stress-induced depressive-like behaviors.These physiological and behavioral changes allow rats to avoid potential deleterious effects of stress that may result from chronically elevated levels of glucocorticosteroids over days.
基金Qinghai Provincial Key R&D and Transformation Plan (No.2021?SF?C21)。
文摘Objective:To explore the effects of CUMS combined with CRS on mouse hippocampal glial cells and synaptic plasticity-related proteins. Methods: Forty mice were randomly divided into normal group (n=20) and model group (n=20). The model group used CUMS combined with CRS to prepare a mouse model of depression for 7 weeks. The behavioral evaluation of the mice at 3 weeks and 7 weeks after modeling was performed by sugar water preference test, open field test and tail suspension test. After the experiment, the samples were collected, and the content of TNF-a in the hippocampus of mice was detected by enzyme-linked immunosorbent assay. Immunohistochemical method was used to detect the Iba-1 and GFAP MOD values of mouse hippocampal CA1 area, CA3 area and DG area. Western blot was used to detect the protein expression of Iba-1, GFAP, SYN1 and PSD-95 in the hippocampus. fluorescence quantitative PCR method was used to detect the expression of SYN1, PSD-95 mRNA in hippocampus. Results: At the 3rd week after modeling, the body weight, sugar water preference rate, total distance moved, number of standing uprights, and stay time in the central area of the mice in the model group were all lower than those in the normal group (P<0.05), and the tail suspension immobility time was longer than that in the normal group (P<0.01). After 7 weeks of modeling, the body weight, sugar water preference rate, total distance moved, number of erection times, central area residence time, and average movement speed of the mice in the model group were lower than those in the normal group (P< 0.05), the tail suspension immobility time was longer than that in the normal group (P<0.01). The contents of TNF-a in the hippocampus were higher than those in the normal group (P<0.05). The GFAP MOD value and the relative expression of GFAP protein in hippocampal CA1, CA3 and DG regions were significantly lower than those in the normal group (P<0.05). The Iba-1 MOD value and the relative expression of Iba-1 protein in hippocampal CA1, CA3 and DG regions were significantly higher than those in the normal group (P<0.05). The relative expression of SYN1 and PSD-95 protein and the relative expression of SYN1 and PSD-95 mRNA in the hippocampus were significantly lower than those in the normal group (P<0.05). Conclusion: After 3 weeks of CUMS and CRS modeling, the depression-like behavior of mice appeared, and the depression of mice was more obvious after 7 weeks of modeling. The depression mouse model made by CUMS combined with CRS method may be related to increased hippocampal inflammation, excessive activation of microglia, decreased number of astrocytes and decreased synaptic plasticity.
文摘目的 探究肝郁脾虚证的生物学基础及逍遥散的作用机制。方法 采用慢性束缚应激方法建立肝郁脾虚证大鼠模型。72只SD雄性大鼠随机分为6组,正常组,模型组,氟西汀组(0.0018 g/kg),逍遥散高(16.7 g/kg)、中(8.35 g/kg)、低(4.175g/kg)剂量组,每组12只。造模结束后,ELISA法测定大鼠血清单丝氨酸蛋白激酶1(single serine protein kinase 1,LIMK1)的含量,荧光定量PCR、 Western blot检测海马组织RAS同源基因家族成员A(RAS homologous gene family member A,RhoA)、Rho相关螺旋卷曲蛋白激酶2(Rho-related spiral coiled protein kinase 2,ROCK2)的m RNA及蛋白表达。结果 模型组大鼠血清LIMK1含量,海马组织中RhoA、ROCK2的mRNA和蛋白表达水平均明显高于正常组(P<0.01);各药物组大鼠血清LIMK1含量明显低于模型组(P<0.01),逍遥散高、中剂量组和氟西汀组大鼠海马组织中RhoA、ROCK2的mRNA和蛋白表达明显低于模型组(P<0.05,P<0.01)。结论 慢性束缚应激肝郁脾虚证大鼠海马组织RhoA/ROCK2通路被激活,逍遥散可抑制RhoA/ROCK2通路激活,并且作用呈剂量依赖性。
