慢性肝损伤合并应激抑郁大鼠模型的建立及养肝解郁颗粒干预作用的研究
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摘要
目的:用40%CCL4皮下注射(12W)、孤养及慢性不可预见性的温和刺激(3W)的复合造模方法,制备慢性肝损伤合并应激抑郁动物模型,探索慢性肝损伤合并应激抑郁大鼠模型制备的方法;同时观察中药养肝解郁颗粒干预作用,通过观察实验大鼠的肝功能及体重情况,观察行为学改变,并采用高效液相电化学法检测下丘脑神经递质的含量,观察肝脏、大脑海马结构病理改变,以及对大脑海马部位脑源神经神经营养因子(BDNF)、以及下丘脑精氨酸加压素(AVP)mRNA的表达,初步探讨养肝解郁颗粒治疗慢性肝炎合并抑郁症的作用机理,进一步明确该药的保肝、降酶、抗抑郁的作用,并从分子水平说明养肝解郁颗粒保肝抗抑郁的作用机理。为研究慢性肝炎合并抑郁症实验研究奠定基础。
方法:1.将大鼠随机分为8组:空白组、肝损伤模型组(模型1组)、肝损伤合并应激抑郁模型组(模型2组)、乙肝益气解郁颗粒组(对照1组)、盐酸氟西汀胶囊组(对照2组)、养肝解郁颗粒低、中、高剂量组(低、中、高剂量组)。2.予大鼠四氯化碳油溶液皮下注射(1ml/kg,2次/周、共12周),合并孤养及慢性不可预见性的温和刺激的复合造模方法,建立慢性肝损伤合并应激抑郁的大鼠模型。3.测量各组大鼠的体重、糖水消耗量、敞箱试验得分,肝功能改变、下丘脑内神经递质的含量,以及肝脏及大脑海马部位的病理改变、大脑海马部位CA1区BDNF神经阳性细胞表达及下丘脑AVP-mRNA的表达。
结果:40%CCL4油溶液,(1ml/kg,2次/周、共12周),合并孤养及慢性不可预见性的温和刺激的复合造模方法能成功制备慢性肝损伤合并应激抑郁大鼠模型;在行为学上对慢性肝损伤合并应激抑郁大鼠的影响,养肝解郁颗粒能够增加模型体重、提高糖水偏爱百分比及敞箱实验得分,且疗效优于对照1及对照2组(P <0.05);在肝功能慢性肝损伤合并应激抑郁大鼠的影响,养肝解郁颗粒对肝功能各相指标的影响与对照1组及对照2组比较(P <0.01),说明养肝解郁颗粒可以降低升高的转氨酶及黄疸,升高降低的蛋白,且疗效优于对照1组及对照2组;对下丘脑神经递质含量的影响,养肝解郁颗粒与对照1组比较,优于对照1组(P <0.01);与对照2组比较,疗效相当(P>0.05);对肝脏病理结构的改善,养肝解郁颗粒优于对照1组及对照2组(P <0.01),能够减少肝脏细胞的坏死;对改善大鼠大脑海马部位的结构的影响,治疗高、中剂量组明显优于对照1组(P <0.01),与对照2组相比,疗效相当(P>0.05);对大脑海马部位BDNF阳性神经元表达的影响,治疗高、中剂量组明显优于对照1组(P <0.05),与对照2组比较,疗效相当(P>0.05);对下丘脑AVP-mRNA表达的影响,治疗高、中剂量组疗效明显优于对照1组(P <0.05),与对照2组比较,高、中剂量组疗效相当(P>0.05)。
结论:1.采用四氯化碳油溶液皮下注射(2次/周、共12周),合并孤养及慢性不可预见性的温和刺激的复合造模方法可成功制备慢性肝损伤合并应激抑郁动物模型。2.本研究所测定各项指标的结果显示:养肝解郁颗粒可以调节模型大鼠的体重、行为学,改善肝功能、神经递质含量,减少肝脏细胞的坏死,改善大鼠大脑海马部位的结构及增加CA1区BDNF阳性神经元的表达,降低下丘脑AVP-mRNA的表达,在肝病治疗上其疗效较优于乙肝益气解郁颗粒,在抑郁症治疗上与盐酸氟西汀胶囊疗效相当。3.养肝解郁颗粒具有保肝降酶抗抑郁的作用,值得进一步研究和开发。
Objective :Contrast with 40% CCL4 subcutaneous (12W), isolated and dependent chronic unpredictable mild stimulation (3W) model of the composite method of preparation of the combined stress of chronic liver injury animal model of depression, chronic liver injury to explore the combined stress-induced depression great Preparation of mouse model of the method. At the same time observe the effect of intervention Yanggan role Jieyu particles through the observation of the experimental rats liver function and weight, the observed changes in behavior, and electrochemical detection using high-performance liquid hypothalamic neurotransmitter levels were observed in liver, brain Sea pathological changes in the structure of horse, as well as parts of the brain hippocampus brain-derived neurotrophic factor nerve (BDNF), as well as hypothalamic arginine vasopressin (AVP) mRNA expression, Preliminary study Jieyu Yanggan particles combined treatment of chronic hepatitis mechanism of depression, further clarified the drug's liver, Jiangmei, the role of antidepressants and explain the molecular level from particles Yanggan Jieyu antidepressant mechanism of liver . Combined to study depression in chronic hepatitis lay the foundation for experimental research.
Methods:1.Dividatur the rats into 8 groups randamly:blank group ,chronic hepatic injury model group (model group 1),chronic hepatic injury and stress depression model group (model group 2),Yigan Yiqi Jieyu Keli group ( control 1 group ),Fluoxetine Hydrochloride Capsules group (control 2 group ),the diminutive、midst、large dose of Yanggan Jieyu Keli(Treatment of diminutive、midst、large -dose group).2.The model was established by subcutaneous injection of carbon tetrachloride and separate bree with CUMS. The rats' weight,the Open-field test,sucrose intake, monoamine neurotransmitters in hypothalamus of rats were measured during the experiment,and liver pathology was observed.3. Measurement of body weight of rats in each group,dessert consumption, open-field test scores,liver function changes in hypothalamic neurotransmitters in the content, as well as liver and brain pathological changes in the hippocampus,the brain area CA1 hippocampus neurons BDNF positive cells and expression of hypothalamic AVP-mRNA expression.
Results: Contrast with 40% CCL4 oil solution subcutaneous (12W), (1ml/kg, 2 times / week for 12 weeks), the merger raised alone and chronic unpredictable mild stimulation of the composite model can successfully merge the preparation of chronic liver injury in rat model of stress-induced depression; Study on the behavior of chronic liver injury with the impact of stress-induced depression in rats, Treatment of diminutive、midst-dose group can increase weight, improve the percentage of preference for dessert and open box test scores, and more effective than the control group 1 and 2 (P <0.05); Liver function of chronic stress-induced depression of liver injury in rats, the liver function of the target ,And comparison with control group1 and the control group 2 (P <0.01), Treatment of diminutive、midst-dose group that can reduce the elevated transaminases and jaundice, elevated to reduce the protein, And one more effective than the control group and control group 2. On the hypothalamic content of neurotransmitters, Treatment of diminutive、midst-dose group with the control group1, one better than the control group 1 (P <0.01), with the control group2, a considerable effect (P > 0.05); To improve the structure of the liver pathology, Treatment of diminutive、midst-dose group is better than the control group1 and control group 2 (P <0.01), be able to reduce liver cell necrosis; To improve the rat hippocampus of the brain structure,Treatment of diminutive、midst-dose group was significantly better than the control group 1 (P <0.01), compared with the control group 2, significant effect (P > 0.05); Hippocampus of the brain-positive neurons in BDNF expression, Treatment of diminutive、midst-dose group was significantly better than the control group 1 (P <0.05), compared with the control group 2, significant effect (P > 0.05); On the hypothalamic AVP-mRNA expression,Treatment of diminutive、midst dose group was superior to the control efficacy of one group (P <0.05), compared with the control group 2, diminutive、midst dose group a considerable effect (P > 0.05)。
Conclusion:We can produce the chronic liver injury and separate bree with CUMS model of rats successfully by subcutaneous injection of carbon tetrachloride and separate bree with CUMS. 2. Yanggan Jieyu keli can adjust the weight, behavior, improve liver function, neurotransmitter content, reduce necrosis of liver cells, rat brain to improve the structure of hippocampus CA1 area and increase the BDNF-positive neurons in the expression of to reduce the hypothalamic AVP-mRNA expression. At its effect on the treatment of liver diseases of yigan Yiqi Jieyu keli more than particles in the treatment of depression with fluoxetine hydrochloride capsules considerable effect. In the treatment of depression with significant effects of fluoxetine hydrochloride capsules.3 Yanggan Jieyu Keli had the effect of liver protective and antidepressant, and is worthy of further study and development.
方法:1.将大鼠随机分为8组:空白组、肝损伤模型组(模型1组)、肝损伤合并应激抑郁模型组(模型2组)、乙肝益气解郁颗粒组(对照1组)、盐酸氟西汀胶囊组(对照2组)、养肝解郁颗粒低、中、高剂量组(低、中、高剂量组)。2.予大鼠四氯化碳油溶液皮下注射(1ml/kg,2次/周、共12周),合并孤养及慢性不可预见性的温和刺激的复合造模方法,建立慢性肝损伤合并应激抑郁的大鼠模型。3.测量各组大鼠的体重、糖水消耗量、敞箱试验得分,肝功能改变、下丘脑内神经递质的含量,以及肝脏及大脑海马部位的病理改变、大脑海马部位CA1区BDNF神经阳性细胞表达及下丘脑AVP-mRNA的表达。
结果:40%CCL4油溶液,(1ml/kg,2次/周、共12周),合并孤养及慢性不可预见性的温和刺激的复合造模方法能成功制备慢性肝损伤合并应激抑郁大鼠模型;在行为学上对慢性肝损伤合并应激抑郁大鼠的影响,养肝解郁颗粒能够增加模型体重、提高糖水偏爱百分比及敞箱实验得分,且疗效优于对照1及对照2组(P <0.05);在肝功能慢性肝损伤合并应激抑郁大鼠的影响,养肝解郁颗粒对肝功能各相指标的影响与对照1组及对照2组比较(P <0.01),说明养肝解郁颗粒可以降低升高的转氨酶及黄疸,升高降低的蛋白,且疗效优于对照1组及对照2组;对下丘脑神经递质含量的影响,养肝解郁颗粒与对照1组比较,优于对照1组(P <0.01);与对照2组比较,疗效相当(P>0.05);对肝脏病理结构的改善,养肝解郁颗粒优于对照1组及对照2组(P <0.01),能够减少肝脏细胞的坏死;对改善大鼠大脑海马部位的结构的影响,治疗高、中剂量组明显优于对照1组(P <0.01),与对照2组相比,疗效相当(P>0.05);对大脑海马部位BDNF阳性神经元表达的影响,治疗高、中剂量组明显优于对照1组(P <0.05),与对照2组比较,疗效相当(P>0.05);对下丘脑AVP-mRNA表达的影响,治疗高、中剂量组疗效明显优于对照1组(P <0.05),与对照2组比较,高、中剂量组疗效相当(P>0.05)。
结论:1.采用四氯化碳油溶液皮下注射(2次/周、共12周),合并孤养及慢性不可预见性的温和刺激的复合造模方法可成功制备慢性肝损伤合并应激抑郁动物模型。2.本研究所测定各项指标的结果显示:养肝解郁颗粒可以调节模型大鼠的体重、行为学,改善肝功能、神经递质含量,减少肝脏细胞的坏死,改善大鼠大脑海马部位的结构及增加CA1区BDNF阳性神经元的表达,降低下丘脑AVP-mRNA的表达,在肝病治疗上其疗效较优于乙肝益气解郁颗粒,在抑郁症治疗上与盐酸氟西汀胶囊疗效相当。3.养肝解郁颗粒具有保肝降酶抗抑郁的作用,值得进一步研究和开发。
Objective :Contrast with 40% CCL4 subcutaneous (12W), isolated and dependent chronic unpredictable mild stimulation (3W) model of the composite method of preparation of the combined stress of chronic liver injury animal model of depression, chronic liver injury to explore the combined stress-induced depression great Preparation of mouse model of the method. At the same time observe the effect of intervention Yanggan role Jieyu particles through the observation of the experimental rats liver function and weight, the observed changes in behavior, and electrochemical detection using high-performance liquid hypothalamic neurotransmitter levels were observed in liver, brain Sea pathological changes in the structure of horse, as well as parts of the brain hippocampus brain-derived neurotrophic factor nerve (BDNF), as well as hypothalamic arginine vasopressin (AVP) mRNA expression, Preliminary study Jieyu Yanggan particles combined treatment of chronic hepatitis mechanism of depression, further clarified the drug's liver, Jiangmei, the role of antidepressants and explain the molecular level from particles Yanggan Jieyu antidepressant mechanism of liver . Combined to study depression in chronic hepatitis lay the foundation for experimental research.
Methods:1.Dividatur the rats into 8 groups randamly:blank group ,chronic hepatic injury model group (model group 1),chronic hepatic injury and stress depression model group (model group 2),Yigan Yiqi Jieyu Keli group ( control 1 group ),Fluoxetine Hydrochloride Capsules group (control 2 group ),the diminutive、midst、large dose of Yanggan Jieyu Keli(Treatment of diminutive、midst、large -dose group).2.The model was established by subcutaneous injection of carbon tetrachloride and separate bree with CUMS. The rats' weight,the Open-field test,sucrose intake, monoamine neurotransmitters in hypothalamus of rats were measured during the experiment,and liver pathology was observed.3. Measurement of body weight of rats in each group,dessert consumption, open-field test scores,liver function changes in hypothalamic neurotransmitters in the content, as well as liver and brain pathological changes in the hippocampus,the brain area CA1 hippocampus neurons BDNF positive cells and expression of hypothalamic AVP-mRNA expression.
Results: Contrast with 40% CCL4 oil solution subcutaneous (12W), (1ml/kg, 2 times / week for 12 weeks), the merger raised alone and chronic unpredictable mild stimulation of the composite model can successfully merge the preparation of chronic liver injury in rat model of stress-induced depression; Study on the behavior of chronic liver injury with the impact of stress-induced depression in rats, Treatment of diminutive、midst-dose group can increase weight, improve the percentage of preference for dessert and open box test scores, and more effective than the control group 1 and 2 (P <0.05); Liver function of chronic stress-induced depression of liver injury in rats, the liver function of the target ,And comparison with control group1 and the control group 2 (P <0.01), Treatment of diminutive、midst-dose group that can reduce the elevated transaminases and jaundice, elevated to reduce the protein, And one more effective than the control group and control group 2. On the hypothalamic content of neurotransmitters, Treatment of diminutive、midst-dose group with the control group1, one better than the control group 1 (P <0.01), with the control group2, a considerable effect (P > 0.05); To improve the structure of the liver pathology, Treatment of diminutive、midst-dose group is better than the control group1 and control group 2 (P <0.01), be able to reduce liver cell necrosis; To improve the rat hippocampus of the brain structure,Treatment of diminutive、midst-dose group was significantly better than the control group 1 (P <0.01), compared with the control group 2, significant effect (P > 0.05); Hippocampus of the brain-positive neurons in BDNF expression, Treatment of diminutive、midst-dose group was significantly better than the control group 1 (P <0.05), compared with the control group 2, significant effect (P > 0.05); On the hypothalamic AVP-mRNA expression,Treatment of diminutive、midst dose group was superior to the control efficacy of one group (P <0.05), compared with the control group 2, diminutive、midst dose group a considerable effect (P > 0.05)。
Conclusion:We can produce the chronic liver injury and separate bree with CUMS model of rats successfully by subcutaneous injection of carbon tetrachloride and separate bree with CUMS. 2. Yanggan Jieyu keli can adjust the weight, behavior, improve liver function, neurotransmitter content, reduce necrosis of liver cells, rat brain to improve the structure of hippocampus CA1 area and increase the BDNF-positive neurons in the expression of to reduce the hypothalamic AVP-mRNA expression. At its effect on the treatment of liver diseases of yigan Yiqi Jieyu keli more than particles in the treatment of depression with fluoxetine hydrochloride capsules considerable effect. In the treatment of depression with significant effects of fluoxetine hydrochloride capsules.3 Yanggan Jieyu Keli had the effect of liver protective and antidepressant, and is worthy of further study and development.
引文
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