基于正常大鼠物质能量代谢的栀子不同炮制品寒热药性研究

Study on Cold and Heat Properties of Raw Gardeniae Fructus,Stir-Baked Gardeniae Fructus,and Gardeniae Fructus Praeparatus Based on Substance and Energy Metabolism in Normal Rats

目的通过检测栀子不同炮制品对正常大鼠物质代谢与能量代谢水平的影响,以期阐明炒制引起栀子寒热药性变化的科学内涵。方法雄性SD大鼠随机分为对照组、生栀子组、炒栀子组及焦栀子组。对照组大鼠灌胃蒸馏水(1 mL·100 g^(-1)),生栀子组、炒栀子组及焦栀子组大鼠分别灌胃给予等体积的相应炮制品水提取物,连续给药9天。记录各组大鼠给药期间体质量、肛温变化。酶联免疫吸附(ELISA)法检测大鼠肝脏组织中物质能量代谢相关指标GCK、PFK、PK、Acetyl-CoA、PDH、CS、ICD、α-KGDHC、PYGL、GSK-3、ATGL、CCR、COX、ATPs、ADK、Na^(+)-K^(+)-ATP酶、ATP、ADP、NAD及NADH的含量。免疫组化法(Immunohistochemistry,IHC)检测AMPK/mTOR信号通路关键蛋白的表达。结果与对照组相比,生栀子极显著性降低大鼠体质量、肛温、PK、PDH、Acetyl-CoA、CS、ICD、PYGL、GSK-3、ATGL、CCR、ATPs、ADK、Na^(+)-K^(+)-ATP酶、ATP、NAD^(+)、NADH、AMPK(P<0.01),显著性降低大鼠GCK、PFK、α-KGDHC、COX、ADP水平(P<0.05),极显著升高大鼠mTOR水平(P<0.01);炒栀子极显著性降低大鼠体质量、肛温、PK、PDH、Acetyl-CoA、CS、ICD、PYGL、ATGL、ATPs、ADK、Na^(+)-K^(+)-ATP酶、ATP、NAD^(+)、NADH水平(P<0.01),显著性降低大鼠PFK、α-KGDHC、GSK-3、COX、ADP及AMPK水平(P<0.05),极显著升高大鼠mTOR水平(P<0.01);焦栀子极显著性降低大鼠肛温、PK、Acetyl-CoA、CS、ICD、PYGL、ATGL、ATPs、ADK、NAD^(+)水平(P<0.01),显著降低大鼠体质量、PDH、Na^(+)-K^(+)-ATP酶、ATP及NADH(P<0.05),极显著升高大鼠mTOR水平(P<0.01)。与生栀子组相比,炒栀子极显著性升高大鼠PK、PDH、Acetyl-CoA、ICD、ADK及NADH水平(P<0.01),显著性升高大鼠PDH水平(P<0.05);焦栀子极显著性升高大鼠PK、PDH、Acetyl-CoA、CS、ICD、ADK、Na^(+)-K^(+)-ATP酶、NAD^(+)及NADH水平(P<0.01),显著性升高大鼠GSK-3、ATPs及AMPK水平(P<0.05)。与炒栀子组相比,焦栀子可极显著性升高大鼠Na^(+)-K^(+)-ATP酶及NADH水平(P<0.01)。结论生栀子组、炒栀子组及焦栀子组均能显著降低正常大鼠物质能量代谢水平,说明三者均属于寒性,这与《中国药典》记载的生栀子、炒栀子、焦栀子均属寒性一致。就大鼠物质能量代谢而言,焦栀子组高于炒栀子组,炒栀子组高于生栀子组,说明焦栀子寒性弱于炒栀子,炒栀子寒性弱于生栀子。

Methods The male SD rats were randomly divided into control group,raw Gardeniae Fructus group,stirbaked Gardeniae Fructus group,and Gardeniae Fructus Praeparatus group.The rats of raw Gardeniae Fructus group,stir-baked Gardeniae Fructus group,and Gardeniae Fructus Praeparatus group were administrated with the water extract of raw Gardeniae Fructus,stir-baked Gardeniae Fructus and Gardeniae Fructus Praeparatus,respectively,and the rats of control group were given the normal saline for 9 days.The changes of body weight and anal temperature during administration were record.Enzyme linked immunosorbent assay(ELISA)was used to detect the expression levels of GCK,PFK,PK,Acetyl-CoA,PDH,CS,ICD,α-KGDHC,PYGL,GSK-3,ATGL,CCR,COX,ATPs,ADK,Na^(+)-K^(+)-ATPase,ATP,ADP,NAD^(+),and NADH.The expression of key proteins in AMPK/mTOR signaling pathway were detected by immunohistochemistry method.Results Compared with the control group,raw Gardeniae Fructus significantly reduced the body weight,anal temperature,the levels of PK,PDH,Acetyl-CoA,CS,ICD,PYGL,GSK-3,ATGL,CCR,ATPs,ADK,Na^(+)-K^(+)-ATPase,ATP,NAD^(+),NADH,AMPK,significantly elevated the level of mTOR(P<0.01),the levels of GCK,PFK,α-KGDHC,COX,ADP(P<0.05),stir-baked Gardeniae Fructus significantly reduced the body weight,anal temperature,the levels of PK,PDH,Acetyl-CoA,CS,ICD,PYGL,ATGL,ATPs,ADK,Na^(+)-K^(+)-ATPase,ATP,NAD^(+),NADH,significantly elevated the level of mTOR(P<0.01),the levels of PFK,α-KGDHC,GSK-3,COX,ADP,AMPK(P<0.05),Gardeniae Fructus Praeparatus significantly reduced the anal temperature,the levels of PK,Acetyl-CoA,CS,ICD,PYGL,ATGL,ATPs,ADK,NAD^(+),significantly elevated the level of mTOR(P<0.01),the body weight,the levels of PDH,Na^(+)-K^(+)-ATPase,ATP,NADH(P<0.05).Compared with raw Gardeniae Fructus group,stir-baked Gardeniae Fructus significantly increased the levels of PK,PDH,Acetyl-CoA,ICD,ADK,NADH(P<0.01),the level of PDH(P<0.05).Gardeniae Fructus Praeparatus significantly increased the levels of PK,PDH,Acetyl-CoA,CS,ICD,ADK,Na^(+)-K^(+)-ATPase,NAD^(+),NADH(P<0.01),the levels of GSK-3,ATPs,AMPK(P<0.05).Compared with stir-baked Gardeniae Fructus group,Gardeniae Fructus Praeparatus significantly increased the levels of Na^(+)-K^(+)-ATPase,NADH(P<0.01).Conclusion The raw Gardeniae Fructus,stir-baked Gardeniae Fructus,and Gardeniae Fructus Praeparatus could significantly reduce the levels of substance and energy metabolism,indicating that the three processed products of Gardenia Fructus all showed the cold property,which was consistent with the record in Chinese Pharmacopoeia.The levels of substance and energy metabolism of rats in the Gardeniae Fructus Praeparatus group were significantly higher than those of rats in the stir-baked Gardeniae Fructus group,while the levels of substance and energy metabolism of rats in the stir-baked Gardeniae Fructus group were significantly higher than those of rats in the raw Gardeniae Fructus,suggesting that the cold property of the Gardeniae Fructus Praeparatus was weaker than that of the stir-baked Gardeniae Fructus and the cold property of the stir-baked Gardeniae Fructus was weaker than that of raw Gardeniae Fructus.Obiective To clarify the scientific connotation of the changes in cold and heat properties of raw Gardeniae Fructus,stir-baked Gardeniae Fructus,and Gardeniae Fructus Praeparatus based on substance and energy metabolism after drug intervention in the normal rats.