益气活血中药配伍对慢性间歇性低氧复合胰岛素抵抗介导的动脉粥样硬化动物模型的干预效应
|
摘要
该文研究益气活血中药配伍对间歇性低氧(CIH)复合胰岛素抵抗(IR)介导的动脉粥样硬化(AS)小鼠模型的干预效应,并观察SREBP-1c相关信号分子在其中的作用。实验采用高脂饮食诱导联合STZ注射法建立IR的Apo E-/-小鼠模型,以低氧动物饲养仪处理IR-ApoE-/-小鼠模型,每日处理8 h,连续处理8周,以建立CIH+IR-ApoE-/-小鼠模型。模型小鼠随机分为7组:常氧对照组(NC)、模型组(CIH)、SREBPs抑制剂组(betulin)、阿托伐他汀组(WM)、益气活血中药低剂量组(TCML)、益气活血中药中剂量组(TCM-M)、益气活血中药高剂量组(TCM-H),同时常规饲养的C57BL/6J小鼠10只作为空白对照组(Con)。观察益气活血中药配伍(人参皂苷、川芎嗪)对模型小鼠IR,血脂水平,炎症反应,氧化应激及SREBP-1c相关信号分子的干预效应。结果显示,TCM-H组血糖低于其他实验组(P<0.05);betulin组、WM组、TCM-H组HDL-C高于CIH组(P<0.05),TCM-M组、TCM-H组LDL-C低于CIH组(P<0.05);CIH组血清CRP水平高于其他各组(P<0.05);TCM-H组SOD水平较CIH组升高(P<0.05);NC组、CIH组小鼠主动脉可见明显的AS斑块,betulin组、WM组、TCM-H组AS斑块减小;主动脉降段,WM组及TCM-H组AS斑块较CIH组减少且面积较小;TCM-H组主动脉、骨骼肌SREBP-1c和FAS的mRNA及蛋白表达水平均较CIH组降低(P<0.05);TCM-H组TNF-α和CD106(VCAM-1)蛋白表达水平较CIH组降低(P<0.05);TCM-H组主动脉、骨骼肌、肝组织p-IRS-1的蛋白表达水平较CIH组升高(P<0.05);TCM-H组主动脉、肝HIF-1α蛋白表达水平较CIH组降低(P<0.05)。TCM-H组血糖低于其他实验组;CIH组HDL-C低于其他各组,TCM-H组LCL-C低于CIH组;CIH组CRP水平高于其他各组;TCM-H组SOD水平高于CIH组;NC组及CIH组AS斑块较其他组明显增多;在主动脉降段,WM组及TCMH组AS斑块较CIH组减少且面积较小;在主动脉及骨骼肌,TCM-H组SREBP-1c和FAS的mRNA及蛋白表达水平均较CIH组降低;TCM-H组TNF-α和CD106(VCAM-1)蛋白表达水平较CIH组降低;在主动脉、骨骼肌及肝组织,TCM-H组p-IRS-1的蛋白表达水平较CIH组升高;在主动脉及肝组织,TCM-H组HIF-1α蛋白表达水平较CIH组降低。该研究表明,人参皂苷、TMP配伍能够一定程度改善间歇性低氧复合IR的动脉粥样硬化模型IR及血脂水平,降低炎症反应和氧化应激程度,并最终抑制AS进展,其机制可能与抑制SREBP-1c相关信号分子的表达水平有关。
This study aims to observe the intervention effects of Chinese herbal medicine of supplementing Qi and activating blood circulation on chronic intermittent hypoxia(CIH) composite insulin resistance(IR) mediated atherosclerosis(AS) mice model,and to observe the mechanism of SREBP-1 c signaling molecule.IR Apo E-/-mice model was induced by high-fat diet combined with STZ injection.Then the mice were treated with hypoxic animal incubator for 8 h per day and 8 weeks to establish a CIH+IR-ApoE-/-mouse model.Model mice were randomly and averagely divided into normoxic control group(NC),model group(CIH) and SREBPs inhibitor group(betulin),atorvastatin group(WM),TCM low-dose group(TCM-L),TCM middle-dose group(TCM-M) and TCM high-dose group(TCM-H) group.Chinese herbal medicine of supplementing Qi and activating blood circulation including ginsenosides combined with ligustrazine(TMP) were used as intervention drugs.The study observed the effect of drugs on IR,serum lipid,inflammation,stress,AS and SREBP-1 c related molecules.The results showed that fasting blood glucose in TCM-H group decreased compared with other experimental groups(P<0.05).HDL-C level in betulin group,WM group,TCM-H group was higher than that in CIH group(P<0.05).LDL-C level in TCM-M group,TCM-H group is lower than that in CIH group(P<0.05).The level of CRP in CIH group was higher than that in other groups(P<0.05).The level of SOD in TCM-H group was higher than that in CIH group(P<0.05).NC group and CIH group showed obvious AS aortic plaque,while betulin group,WM group,TCM-H group showed reduction in AS plaque(P<0.05).For descending aorta,AS plaque in CIH group was multiple and large,while less and smaller in WM group and TCM-H(P<0.05).The expression of SREBP-1 c and FAS in aorta and skeletal muscle in TCM-H group was lower than that in CIH group(P<0.05).In aorta,the expression of TNF-α and CD106(VCAM-1) was lower in TCM-H group than that in CIH group(P<0.05).In aorta,skeletal muscle and liver,the level of p-IRS-1 in TCM-H group was significantly higher than that in CIH group(P<0.05).In aorta and liver,the expression of HIF-1α in TCM-H group was lower than that in CIH group(P<0.05).The study demonstrated that combination ginsenosides with TMP could improve IR and serum lipid level and inhibit inflammation and oxidative stress as well as ultimately alleviate AS to some extent.And the mechanism of its interventional effects might be related to the inhibition of CIH-induced upregulation of SREBP-1 c related molecules.
This study aims to observe the intervention effects of Chinese herbal medicine of supplementing Qi and activating blood circulation on chronic intermittent hypoxia(CIH) composite insulin resistance(IR) mediated atherosclerosis(AS) mice model,and to observe the mechanism of SREBP-1 c signaling molecule.IR Apo E-/-mice model was induced by high-fat diet combined with STZ injection.Then the mice were treated with hypoxic animal incubator for 8 h per day and 8 weeks to establish a CIH+IR-ApoE-/-mouse model.Model mice were randomly and averagely divided into normoxic control group(NC),model group(CIH) and SREBPs inhibitor group(betulin),atorvastatin group(WM),TCM low-dose group(TCM-L),TCM middle-dose group(TCM-M) and TCM high-dose group(TCM-H) group.Chinese herbal medicine of supplementing Qi and activating blood circulation including ginsenosides combined with ligustrazine(TMP) were used as intervention drugs.The study observed the effect of drugs on IR,serum lipid,inflammation,stress,AS and SREBP-1 c related molecules.The results showed that fasting blood glucose in TCM-H group decreased compared with other experimental groups(P<0.05).HDL-C level in betulin group,WM group,TCM-H group was higher than that in CIH group(P<0.05).LDL-C level in TCM-M group,TCM-H group is lower than that in CIH group(P<0.05).The level of CRP in CIH group was higher than that in other groups(P<0.05).The level of SOD in TCM-H group was higher than that in CIH group(P<0.05).NC group and CIH group showed obvious AS aortic plaque,while betulin group,WM group,TCM-H group showed reduction in AS plaque(P<0.05).For descending aorta,AS plaque in CIH group was multiple and large,while less and smaller in WM group and TCM-H(P<0.05).The expression of SREBP-1 c and FAS in aorta and skeletal muscle in TCM-H group was lower than that in CIH group(P<0.05).In aorta,the expression of TNF-α and CD106(VCAM-1) was lower in TCM-H group than that in CIH group(P<0.05).In aorta,skeletal muscle and liver,the level of p-IRS-1 in TCM-H group was significantly higher than that in CIH group(P<0.05).In aorta and liver,the expression of HIF-1α in TCM-H group was lower than that in CIH group(P<0.05).The study demonstrated that combination ginsenosides with TMP could improve IR and serum lipid level and inhibit inflammation and oxidative stress as well as ultimately alleviate AS to some extent.And the mechanism of its interventional effects might be related to the inhibition of CIH-induced upregulation of SREBP-1 c related molecules.
引文
[1]Wu C X,Liu Y,Zhang J C.Chronic intermittent hypoxia and hypertension:a review of systemic inflammation and Chinese medicine[J].Chin J Integr Med,2013,19(5):394.
[2]Liu Y,Jiang Y,Zhang J,et al.Integrative Chinese and Western medicine on atherosclerosis of coronary heart disease:what are the new control strategies?[J].Chin Sci Bull,2014,59(11):1091.
[3]Ma L Q,Zhang J C,Liu Y.Roles and mechanisms of obstructive sleep apnea-hypopnea syndrome and chronic intermittent hypoxia in atherosclerosis:evidence and prospective[J].Oxid Med Cell Longev,2016,2016:8215082.
[4]Drager L F,Polotsky V Y,Lorenzi-Filho G.Obstructive sleep apnea:an emerging risk factor for atherosclerosis[J].Chest,2011,140(2):534.
[5]刘志国,李磊,国钰妍,等.睡眠呼吸暂停综合征中医证候要素分布与AHI、BMI指数相关性研究[J].世界中西医结合杂志,2013,8(11):1133.
[6]Chai C,Kou J,Zhu D,et al.Mice exposed to chronic intermittent hypoxia simulate clinical features of deficiency of both Qi and Yin syndrome in traditional Chinese medicine[J].Evid Based Complement Alternat Med,2011,2011:356252.
[7]Choi J B,Loredo J S,Norman D,et al.Does obstructive sleep apnea increase hematocrit?[J].Sleep Breath,2006,10(3):155.
[8]冯惠平,冯惠清.阻塞性睡眠呼吸暂停低通气综合征患者血小板活化和纤溶活性的研究[J].中华结核和呼吸杂志,2002,25(9):531.
[9]陈文强,王玉来.基于“脉浊”理论对动脉粥样硬化发病的再认识[J].中医杂志,2013,54(17):1450.
[10]Dai G H,Liu N,Zhu J W,et al.Qi-Shen-Yi-Qi dripping pills promote angiogenesis of ischemic cardiac microvascular endothelial cells by regulating microRNA-223-3p expression[J].Evid Based Complement Alternat Med,2016,2016:5057328.
[11]刘楠,姜云耀,黄婷婷,等.基于网络药理学方法研究补阳还五汤治疗脑梗死的作用机制[J].中国中药杂志,2018,43(11):2190.
[12]沈宁,邱梦茹,朱雪,等.睡眠呼吸暂停低通气综合征中医辨治思路[J].中医杂志,2015,56(4):297.
[13]马林沁,张京春,刘玥,等.中药人参制剂对胰岛素抵抗干预效应的Meta分析[J].中国循证医学杂志,2016,16(7):819.
[14]蒋跃绒,谢元华,张京春,等.陈可冀治疗心血管疾病血瘀证用药规律数据挖掘[J].中医杂志,2015,56(5):376.
[15]陈可冀.川芎嗪的化学、药理与临床应用[M].2版.北京:人民卫生出版社,2014:40.
[16]邬春晓,张京春,刘玥,等.补肾清肝中药配伍对间歇性低氧诱导人脐静脉内皮细胞损伤模型p38MAPK/NF-κB信号通路的干预效应研究[J].中国中药杂志,2017,42(23):4661.
[17]陈泠,李春艳,王政,等.HPLC法测定人参毛状根及不同人参样品中9种人参皂苷的含量[J].药学实践杂志,2016,34(4):313.
[18]赵艳峰.振源胶囊中人参皂苷Re含量测定方法分析[J].中国药物经济学,2014(1):41.
[19]庞威.“振源口服液”的二次开发及其原料药人参果总皂苷的质量研究[D].长春:长春中医药大学,2014.
[20]Mo W L,Chai C Z,Kou J P,et al.Sheng-Mai-San attenuates contractile dysfunction and structural damage induced by chronic intermittent hypoxia in mice[J].Chin J Nat Med,2015,13(10):743.
[21]Sole S S,Srinivasan B P,Akarte A S.Anti-inflammatory action of Tamarind seeds reduces hyperglycemic excursion by repressing pancreaticβ-cell damage and normalizing SREBP-1c concentration[J].Pharm Biol,2013,51(3):350.
[22]Xie Z,Truong T L,Pei Z,et al.Dan-Qi prescription ameliorates insulin resistance through overall corrective regulation of glucose and fat metabolism[J].J Ethnopharmacol,2015,172:70.
[23]Shin H S,Han J M,Kim H G,et al.Anti-atherosclerosis and hyperlipidemia effects of herbal mixture,Artemisia iwayomogi Kitamura and Curcuma longa Linne,in apolipoprotein E-deficient mice[J].J Ethnopharmacol,2014,153(1):142.
[24]吴为.阿托伐他汀在炎症状态下对ApoE-/-小鼠肝脏的影响及其可能机制研究[D].重庆:重庆医科大学,2017.
[25]龚勇珍,姚海伦,孙少卫,等.SREBP-1/小凹蛋白1介导烟酸姜黄素酯对ApoE-/-小鼠的抗动脉粥样硬化作用[J].中国动脉硬化杂志,2014,22(12):1189.
[26]Shimomura I,Bashmakov Y,Ikemoto S,et al.Insulin selectively increases SREBP-1c mRNA in the livers of rats with streptozotocin-induced diabetes[J].Proc Natl Acad Sci USA,1999,96(24):13656.
[2]Liu Y,Jiang Y,Zhang J,et al.Integrative Chinese and Western medicine on atherosclerosis of coronary heart disease:what are the new control strategies?[J].Chin Sci Bull,2014,59(11):1091.
[3]Ma L Q,Zhang J C,Liu Y.Roles and mechanisms of obstructive sleep apnea-hypopnea syndrome and chronic intermittent hypoxia in atherosclerosis:evidence and prospective[J].Oxid Med Cell Longev,2016,2016:8215082.
[4]Drager L F,Polotsky V Y,Lorenzi-Filho G.Obstructive sleep apnea:an emerging risk factor for atherosclerosis[J].Chest,2011,140(2):534.
[5]刘志国,李磊,国钰妍,等.睡眠呼吸暂停综合征中医证候要素分布与AHI、BMI指数相关性研究[J].世界中西医结合杂志,2013,8(11):1133.
[6]Chai C,Kou J,Zhu D,et al.Mice exposed to chronic intermittent hypoxia simulate clinical features of deficiency of both Qi and Yin syndrome in traditional Chinese medicine[J].Evid Based Complement Alternat Med,2011,2011:356252.
[7]Choi J B,Loredo J S,Norman D,et al.Does obstructive sleep apnea increase hematocrit?[J].Sleep Breath,2006,10(3):155.
[8]冯惠平,冯惠清.阻塞性睡眠呼吸暂停低通气综合征患者血小板活化和纤溶活性的研究[J].中华结核和呼吸杂志,2002,25(9):531.
[9]陈文强,王玉来.基于“脉浊”理论对动脉粥样硬化发病的再认识[J].中医杂志,2013,54(17):1450.
[10]Dai G H,Liu N,Zhu J W,et al.Qi-Shen-Yi-Qi dripping pills promote angiogenesis of ischemic cardiac microvascular endothelial cells by regulating microRNA-223-3p expression[J].Evid Based Complement Alternat Med,2016,2016:5057328.
[11]刘楠,姜云耀,黄婷婷,等.基于网络药理学方法研究补阳还五汤治疗脑梗死的作用机制[J].中国中药杂志,2018,43(11):2190.
[12]沈宁,邱梦茹,朱雪,等.睡眠呼吸暂停低通气综合征中医辨治思路[J].中医杂志,2015,56(4):297.
[13]马林沁,张京春,刘玥,等.中药人参制剂对胰岛素抵抗干预效应的Meta分析[J].中国循证医学杂志,2016,16(7):819.
[14]蒋跃绒,谢元华,张京春,等.陈可冀治疗心血管疾病血瘀证用药规律数据挖掘[J].中医杂志,2015,56(5):376.
[15]陈可冀.川芎嗪的化学、药理与临床应用[M].2版.北京:人民卫生出版社,2014:40.
[16]邬春晓,张京春,刘玥,等.补肾清肝中药配伍对间歇性低氧诱导人脐静脉内皮细胞损伤模型p38MAPK/NF-κB信号通路的干预效应研究[J].中国中药杂志,2017,42(23):4661.
[17]陈泠,李春艳,王政,等.HPLC法测定人参毛状根及不同人参样品中9种人参皂苷的含量[J].药学实践杂志,2016,34(4):313.
[18]赵艳峰.振源胶囊中人参皂苷Re含量测定方法分析[J].中国药物经济学,2014(1):41.
[19]庞威.“振源口服液”的二次开发及其原料药人参果总皂苷的质量研究[D].长春:长春中医药大学,2014.
[20]Mo W L,Chai C Z,Kou J P,et al.Sheng-Mai-San attenuates contractile dysfunction and structural damage induced by chronic intermittent hypoxia in mice[J].Chin J Nat Med,2015,13(10):743.
[21]Sole S S,Srinivasan B P,Akarte A S.Anti-inflammatory action of Tamarind seeds reduces hyperglycemic excursion by repressing pancreaticβ-cell damage and normalizing SREBP-1c concentration[J].Pharm Biol,2013,51(3):350.
[22]Xie Z,Truong T L,Pei Z,et al.Dan-Qi prescription ameliorates insulin resistance through overall corrective regulation of glucose and fat metabolism[J].J Ethnopharmacol,2015,172:70.
[23]Shin H S,Han J M,Kim H G,et al.Anti-atherosclerosis and hyperlipidemia effects of herbal mixture,Artemisia iwayomogi Kitamura and Curcuma longa Linne,in apolipoprotein E-deficient mice[J].J Ethnopharmacol,2014,153(1):142.
[24]吴为.阿托伐他汀在炎症状态下对ApoE-/-小鼠肝脏的影响及其可能机制研究[D].重庆:重庆医科大学,2017.
[25]龚勇珍,姚海伦,孙少卫,等.SREBP-1/小凹蛋白1介导烟酸姜黄素酯对ApoE-/-小鼠的抗动脉粥样硬化作用[J].中国动脉硬化杂志,2014,22(12):1189.
[26]Shimomura I,Bashmakov Y,Ikemoto S,et al.Insulin selectively increases SREBP-1c mRNA in the livers of rats with streptozotocin-induced diabetes[J].Proc Natl Acad Sci USA,1999,96(24):13656.