左、右归丸及其拆方对去卵巢大鼠下丘脑及肾上腺TGF-β1、PKA蛋白表达的影响
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摘要
目的:通过检测左、右归丸及其拆方对去卵巢(OVX)大鼠下丘脑、肾上腺TGF-β1、PKA蛋白表达的影响,从分子水平分析其对HPA轴的调控机制。方法:将95只SD雌性大鼠随机分为Control组、SHAM组、OVX组、BJL组、ZGW组、YGW组、BYY组、GTY组、ZYY组。除SHAM、Control组外均进行双侧卵巢切除。各组大鼠灌胃12周后处死,取大鼠下丘脑和肾上腺,用Western blot方法检测其TGF-β1、PKA蛋白的表达,放射免疫技术检测血清中ACTH、COR的表达。结果:与Control组比较,OVX组大鼠血清COR、ACTH表达显著降低(P<0.05),下丘脑及肾上腺中TGF-β1蛋白表达水平显著升高(P<0.05),肾上腺中PKA表达显著降低(P<0.05)。各给药组均能对抗上述改变(P<0.05),其中,YGW组、BYY组对于血清COR、ACTH以及肾上腺TGF-β1的调控超过了ZGW组。结论:左、右归丸及其拆方均能调节OVX大鼠下丘脑、肾上腺TGF-β1和肾上腺PKA蛋白的表达,其中"从阴引阳"的右归丸调控作用最接近正常组。
Objective: Analyze the regulatory mechanism of Zuogui/Yougui Pill and their separated prescriptionson HPA axis at the molecular level by investigating the effect of Zuogui/Yougui Pill and their separated prescriptions on the expression of TGF-β1 and PKA protein in hypothalamus and adrenal of ovariectomized rats. Methods: Ninety-five SD female rats were randomly divided into Control group, SHAM group, OVX group, BJL group, ZGW group, YGW group, BYY group, GTY group and ZYY group. Bilateral ovariectomy was performed except for SHAM group and Control group. The rats in each group were killed after 12 weeks of gavage and took hypothalamus and adrenal of rats. The expressions of TGF-β1 and PKA protein were detected by Western blot and the expressions of ACTH and COR in serum were detected by radioimmunoassay. Results: Compared with Control group, the expression of COR and ACTH in serum in OVX group decreased(P<0.05), the expression of TGF-β1 protein in hypothalamus and adrenal increased(P<0.05) and the expression of PKA in adrenal decreased(P<0.05). Each medication administration group was able to combat these changes(P<0.05). The regulation of YGW group and BYY group on COR, ACTH in serum and TGF-β1 in adrenal was higher than ZGW group. Conclusion: Zuogui/Yougui Pill and their separated prescriptions can regulate the expression of TGF-β1 protein in hypothalamus, adrenal gland and PKA protein in adrenal gland of ovariectomized rats. The regulating effect of Yougui Pill which based on the theory ‘inducing yang from yin' is closest to the normal control group.
Objective: Analyze the regulatory mechanism of Zuogui/Yougui Pill and their separated prescriptionson HPA axis at the molecular level by investigating the effect of Zuogui/Yougui Pill and their separated prescriptions on the expression of TGF-β1 and PKA protein in hypothalamus and adrenal of ovariectomized rats. Methods: Ninety-five SD female rats were randomly divided into Control group, SHAM group, OVX group, BJL group, ZGW group, YGW group, BYY group, GTY group and ZYY group. Bilateral ovariectomy was performed except for SHAM group and Control group. The rats in each group were killed after 12 weeks of gavage and took hypothalamus and adrenal of rats. The expressions of TGF-β1 and PKA protein were detected by Western blot and the expressions of ACTH and COR in serum were detected by radioimmunoassay. Results: Compared with Control group, the expression of COR and ACTH in serum in OVX group decreased(P<0.05), the expression of TGF-β1 protein in hypothalamus and adrenal increased(P<0.05) and the expression of PKA in adrenal decreased(P<0.05). Each medication administration group was able to combat these changes(P<0.05). The regulation of YGW group and BYY group on COR, ACTH in serum and TGF-β1 in adrenal was higher than ZGW group. Conclusion: Zuogui/Yougui Pill and their separated prescriptions can regulate the expression of TGF-β1 protein in hypothalamus, adrenal gland and PKA protein in adrenal gland of ovariectomized rats. The regulating effect of Yougui Pill which based on the theory ‘inducing yang from yin' is closest to the normal control group.
引文
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[7]涂平生,胡黎平,黄自为,等.补肾方剂防治去卵巢大鼠骨丢失作用机制的探讨.中医药导报,2007,13(12):13-14,22
[8]许兵,王萧枫,王冠华,等.补肾活血颗粒对去势大鼠肾虚证的影响及机制研究.中华中医药学刊,2015,33(12):2846-2849
[9]姜春华,钟学礼,沈自尹,等.祖国医学肾的研究.上海:上海科学技术出版社,1964:13
[10]王水华,陈帮明,刘永芳,等.养阴益肾颗粒对糖皮质激素致肾阴虚证大鼠HPA轴的影响.中国实验方剂学杂志,2016,22(11):142-147
[11]王德秀,胡旭光,臧建伟,等.六味地黄汤生物制剂对肾阴虚小鼠HPA轴的调节作用研究.陕西中医,2008,29(3):374-375
[12]蒋志文,夏俊,余裕民,等.牛ACTH受体和TGF-β1的调节作用.中国人兽共患病杂志,1999,15(3):69-71
[2]Siris E S,Miller P D,Barrett-Connor E,et al.Identification and fracture outcomes of undiagnosed low bone mineral density in postmenopausal women:results from the National Osteoporosis Risk Assessment.JAMA,2001,286(22):2815-2822
[3]谢林.骨质疏松症动物模型的选择与思考.安徽中医学院学报,1997,16(4):58-59
[4]孟玥,任艳玲,艾思羽,等.左、右归丸及其拆方对去卵巢大鼠肾脏TGFβ1/Smad1表达的影响.辽宁中医杂志,2016,43(9):1981-1983
[5]孟玥,任艳玲,孙月娇,等.左归丸、右归丸及其拆方对去卵巢骨质疏松症模型大鼠肾脏碱性磷酸酶、骨钙素表达的影响.中医杂志,2016,57(5):423-427
[6]支会英,李恩.他莫西芬对去卵巢骨质疏松大鼠影响的实验研究.中国骨质疏松杂志,2001,7(3):60-62,65
[7]涂平生,胡黎平,黄自为,等.补肾方剂防治去卵巢大鼠骨丢失作用机制的探讨.中医药导报,2007,13(12):13-14,22
[8]许兵,王萧枫,王冠华,等.补肾活血颗粒对去势大鼠肾虚证的影响及机制研究.中华中医药学刊,2015,33(12):2846-2849
[9]姜春华,钟学礼,沈自尹,等.祖国医学肾的研究.上海:上海科学技术出版社,1964:13
[10]王水华,陈帮明,刘永芳,等.养阴益肾颗粒对糖皮质激素致肾阴虚证大鼠HPA轴的影响.中国实验方剂学杂志,2016,22(11):142-147
[11]王德秀,胡旭光,臧建伟,等.六味地黄汤生物制剂对肾阴虚小鼠HPA轴的调节作用研究.陕西中医,2008,29(3):374-375
[12]蒋志文,夏俊,余裕民,等.牛ACTH受体和TGF-β1的调节作用.中国人兽共患病杂志,1999,15(3):69-71