养阴活血方药对糖尿病大鼠肝脏线粒体琥珀酸脱氢酶和三磷酸腺苷酶的影响
|
摘要
目的探讨养阴活血方药对糖尿病大鼠肝脏线粒体琥珀酸脱氢酶(SDH)和三磷酸腺苷酶(ATP酶)活性的影响。方法选取SD大鼠24只,随机分为三组:正常对照组,糖尿病模型组,中药干预组。除正常对照组外,其他两组腹腔注射链脲佐菌素(STZ)造糖尿病模型。正常对照组和糖尿病模型组自由饮水,中药干预组按人与动物间体表面积折算的等效剂量比值灌喂给药。造模成功后10周,股动脉放血,并取肝脏,提取大鼠肝脏线粒体,并测定SDH和ATP酶活性的变化,观察肝细胞线粒体超微结构。结果与正常对照组[(3.91±0.43)U/mgprot]比较,糖尿病模型组大鼠肝脏线粒体SDH[(2.31±0.27)U/mgprot]活性显著降低(P<0.01),Na+-K+-ATP酶[(0.57±0.18)U/mgprot]活性显著低于对照组[(2.39±0.28)U/mgprot](P<0.01),Ca2+-Mg2+-ATP酶[(0.95±0.19)U/mgprot]活性显著低于对照组[(2.18±0.09)U/mgprot](P<0.01),肝组织透射电镜观察:肝细胞线粒体空泡化,线粒体嵴、膜大部分融合、消失,粗面内质网脱颗粒现象明显。与糖尿病模型组相比,中药干预组SDH[(3.39±0.29)U/mgprot]、Na+-K+-ATP酶[(1.70±0.22)U/mgprot]和Ca2+-Mg2+-ATP[(1.75±0.09)U/mgprot]酶活性均有显著升高(P<0.01),肝组织透射电镜观察:细胞线粒体无空泡,损伤程度轻。结论养阴活血方药可有效提高大鼠肝脏线粒体SDH活性、Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性,这可能是养阴活血方药对糖尿病大鼠肝脏线粒体保护作用机制之一。
Objective To discuss the effects of Nourishing Yin and Prompting Blood Flow Recipe on activities of suc cinate dehydrogenase(SDH) and adenosine triphosphatase(ATPase) in diabetic rat liver mitochondria.Methods 24 SD rats were randomly divided into three groups: normal control group,diabetic group and traditional Chinese medicine(TRC) intervention group.Diabetic rats were induced by intraperitonea streptozotocin(STZ) injection.Normal control group and diabetic group were drinking water freely.TRC intervention group were administered by equivalent dose ac cording to surface area conversions ratio of human to animal.After 10 weeks,bloodletting was done from arteria cru ralis,then rat liver mitochondria were extracted to observe their ultrastructure and determine activity changes of SDH and ATPase.Results Compared with normal control group [(3.91±0.43) U/mgprot],activities of SDH [(2.31±0.27) U/mgprot] in rat liver mitochondria of diabetic group decreased significantly(P < 0.01).In diabetic group,activities of Na+K+-ATPase [(0.57±0.18) U/mgprot] and Ca2+-Mg2+-ATPase [(0.95±0.19) U/mgprot] were obviously lower than those of normal control group(P < 0.01),which were respectively(2.39±0.28) U/mgprot and(2.18±0.09) U/mgprot.The following were observed in the hepatic tissue by transmission electron microscope(TEM): hepatic cells mitochondria vacuoliza tion,syzygia and obliteratio of most mitochondria cristae and membrane,obvious degranulation of rough endoplasmic reticulum(RER).Compared with diabetic group,activities of SDH [(3.39 ±0.29) U/mgprot] and Na+-K+-ATPase [(1.70± 0.22) U/mgprot] and Ca2+-Mg2+-ATPase [(1.75±0.09) U/mgprot] in rat liver mitochondria of TRC intervention group rose strikingly(P < 0.01).No vacuolus and light injury degree in the cell mitochondria were observed in the hepatic tissue by TEM.Conclusion Nourishing Yin and Prompting Blood Flow Recipe can effectively boost activities of SDH and Na+-K+-ATPase and Ca2+-Mg2+-ATPase in rat liver mitochondria.This may be one of the protection mechanisms of Nourishing Yin and Prompting Blood Flow Recipe on diabetic rat liver mitochondria.
Objective To discuss the effects of Nourishing Yin and Prompting Blood Flow Recipe on activities of suc cinate dehydrogenase(SDH) and adenosine triphosphatase(ATPase) in diabetic rat liver mitochondria.Methods 24 SD rats were randomly divided into three groups: normal control group,diabetic group and traditional Chinese medicine(TRC) intervention group.Diabetic rats were induced by intraperitonea streptozotocin(STZ) injection.Normal control group and diabetic group were drinking water freely.TRC intervention group were administered by equivalent dose ac cording to surface area conversions ratio of human to animal.After 10 weeks,bloodletting was done from arteria cru ralis,then rat liver mitochondria were extracted to observe their ultrastructure and determine activity changes of SDH and ATPase.Results Compared with normal control group [(3.91±0.43) U/mgprot],activities of SDH [(2.31±0.27) U/mgprot] in rat liver mitochondria of diabetic group decreased significantly(P < 0.01).In diabetic group,activities of Na+K+-ATPase [(0.57±0.18) U/mgprot] and Ca2+-Mg2+-ATPase [(0.95±0.19) U/mgprot] were obviously lower than those of normal control group(P < 0.01),which were respectively(2.39±0.28) U/mgprot and(2.18±0.09) U/mgprot.The following were observed in the hepatic tissue by transmission electron microscope(TEM): hepatic cells mitochondria vacuoliza tion,syzygia and obliteratio of most mitochondria cristae and membrane,obvious degranulation of rough endoplasmic reticulum(RER).Compared with diabetic group,activities of SDH [(3.39 ±0.29) U/mgprot] and Na+-K+-ATPase [(1.70± 0.22) U/mgprot] and Ca2+-Mg2+-ATPase [(1.75±0.09) U/mgprot] in rat liver mitochondria of TRC intervention group rose strikingly(P < 0.01).No vacuolus and light injury degree in the cell mitochondria were observed in the hepatic tissue by TEM.Conclusion Nourishing Yin and Prompting Blood Flow Recipe can effectively boost activities of SDH and Na+-K+-ATPase and Ca2+-Mg2+-ATPase in rat liver mitochondria.This may be one of the protection mechanisms of Nourishing Yin and Prompting Blood Flow Recipe on diabetic rat liver mitochondria.
引文
[1]梁金环,栗彦宁,齐锦生,等.养阴活血方药对糖尿病大鼠肾脏的保护作用及机制探讨[J].中国中药杂志,2008,33(14):1728-1731.
[2]齐锦生,李恩,薛智权,等.滋阴补肾方药对糖尿病大鼠骨中一氧化氮合酶mRNA表达的影响[J].中医杂志,2003,44(5):379.
[3]肖凯,王庆松.PTSD样情感行为异常大鼠脑组织ATP酶活性改变[J].西南国防医药,2002,12(3):204-206.
[4]范颖,李楠,孙云峰,等.黄芪有效部位对糖尿病大鼠Na+-K+-ATP酶活性及AMPK蛋白表达的影响[J].中华中医药杂志,2012,27(10):2660-2663.
[5]金星林,千昌石,杜希臣,等.改良法建立大鼠原发性肝癌变模型与病理形态学研究[J].中国现代医学杂志,2009,19(17):2593-2596.
[6]Lokhande SL,Iyer CM,Shinde RR,et al.A comparative study on the fasting and the postprandial dyslipidaemia in type 2 diabetes mellitus[J].Journal of Clinical&Diagnostic Research,2013,7(4):627-630.
[7]Zhijun W,Jeffrey W,Patrick C.Treating type 2 diabetes mellitus with traditional chinese and indian medicinal herbs[J].Evidence-Based Complementary and Alternative Medicine,2013,(2013):7.
[8]程方平,刘松林,李云海,等.温病湿热证大鼠模型肝线粒体Na+-K+-ATP酶活力变化的对比研究[J].浙江中医药大学学报,2008,32(2):174-175.
[9]孙卫,郑学芝,李丽,等.葛根素对糖尿病大鼠胰腺线粒体氧化应激及ATP酶的影响[J].临床合理用药,2011,4(8c):13-14.
[10]Kumar R.Biochemical changes in erythrocyte membrane in type 2diabetes mellitus[J].Indian J Med Sci,2012,66(5&6):131-135.
[11]Huang S,Millar AH.Succinate dehydrogenase:the complex roles of a simple enzyme[J].Curr Opin Plant Biol,2013,16(3):344-349.
[12]胡建鹏,韩小祥,王键,等.3种中药复方对脑缺血再灌注大鼠脑能量代谢相关酶影响的比较研究[J].中国中西医结合杂志,2007,27(3):231-233.
[13]黄丽萍,周蓉,蒙晓芳,等.寒性中药黄芩对大鼠能量代谢的影响[J].中药材,2010,33(4):575-577.
[14]Zakharchenko MV,Zakharchenko AV,Khunderyakova NV,et al.Burst of succinate dehydrogenase andα-ketoglutarate dehydrogenase activity in concert with the expression of genes coding for respiratory chain proteins underlies short-term beneficial physiological stress in mitochondria[J].Int J Biochem Cell Biol,2013,45(1):190-200.
[15]Piasecka M,Wenda-Rózewicka L,Ogoński T.Computerized analysis of cytochemical reactions for dehydrogenases and oxygraphic studies as methods to evaluate the function of the mitochondrial sheath in rat spermatozoa[J].Andrologia,2001,33(1):1-12.
[16]Ceylan-Isik AF,Fliethman RM,Wold LE,et al.Herbal and traditional Chinese medicine for the treatment of cardiovascular complications in diabetes mellitus[J].Current Diabetes Reviews,2008,4(4):320-328.
[2]齐锦生,李恩,薛智权,等.滋阴补肾方药对糖尿病大鼠骨中一氧化氮合酶mRNA表达的影响[J].中医杂志,2003,44(5):379.
[3]肖凯,王庆松.PTSD样情感行为异常大鼠脑组织ATP酶活性改变[J].西南国防医药,2002,12(3):204-206.
[4]范颖,李楠,孙云峰,等.黄芪有效部位对糖尿病大鼠Na+-K+-ATP酶活性及AMPK蛋白表达的影响[J].中华中医药杂志,2012,27(10):2660-2663.
[5]金星林,千昌石,杜希臣,等.改良法建立大鼠原发性肝癌变模型与病理形态学研究[J].中国现代医学杂志,2009,19(17):2593-2596.
[6]Lokhande SL,Iyer CM,Shinde RR,et al.A comparative study on the fasting and the postprandial dyslipidaemia in type 2 diabetes mellitus[J].Journal of Clinical&Diagnostic Research,2013,7(4):627-630.
[7]Zhijun W,Jeffrey W,Patrick C.Treating type 2 diabetes mellitus with traditional chinese and indian medicinal herbs[J].Evidence-Based Complementary and Alternative Medicine,2013,(2013):7.
[8]程方平,刘松林,李云海,等.温病湿热证大鼠模型肝线粒体Na+-K+-ATP酶活力变化的对比研究[J].浙江中医药大学学报,2008,32(2):174-175.
[9]孙卫,郑学芝,李丽,等.葛根素对糖尿病大鼠胰腺线粒体氧化应激及ATP酶的影响[J].临床合理用药,2011,4(8c):13-14.
[10]Kumar R.Biochemical changes in erythrocyte membrane in type 2diabetes mellitus[J].Indian J Med Sci,2012,66(5&6):131-135.
[11]Huang S,Millar AH.Succinate dehydrogenase:the complex roles of a simple enzyme[J].Curr Opin Plant Biol,2013,16(3):344-349.
[12]胡建鹏,韩小祥,王键,等.3种中药复方对脑缺血再灌注大鼠脑能量代谢相关酶影响的比较研究[J].中国中西医结合杂志,2007,27(3):231-233.
[13]黄丽萍,周蓉,蒙晓芳,等.寒性中药黄芩对大鼠能量代谢的影响[J].中药材,2010,33(4):575-577.
[14]Zakharchenko MV,Zakharchenko AV,Khunderyakova NV,et al.Burst of succinate dehydrogenase andα-ketoglutarate dehydrogenase activity in concert with the expression of genes coding for respiratory chain proteins underlies short-term beneficial physiological stress in mitochondria[J].Int J Biochem Cell Biol,2013,45(1):190-200.
[15]Piasecka M,Wenda-Rózewicka L,Ogoński T.Computerized analysis of cytochemical reactions for dehydrogenases and oxygraphic studies as methods to evaluate the function of the mitochondrial sheath in rat spermatozoa[J].Andrologia,2001,33(1):1-12.
[16]Ceylan-Isik AF,Fliethman RM,Wold LE,et al.Herbal and traditional Chinese medicine for the treatment of cardiovascular complications in diabetes mellitus[J].Current Diabetes Reviews,2008,4(4):320-328.