通窍活血汤对血管性痴呆大鼠海马CA1区自噬相关蛋白LC3及Beclin-1的影响
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摘要
目的:探讨通窍活血汤对血管性痴呆大鼠海马CA 1区自噬相关蛋白LC3及Beclin-1表达的影响。方法:50只健康雄性SD实验大鼠随机分为假手术组(Sham组)、血管性痴呆模型组(VD组)、通窍活血汤组高、中、低剂量组,采用改良Pulsinelli四血管阻断法制备血管性痴呆大鼠模型,用Morris水迷宫实验检测模型,4周后采用免疫印迹法检测各组大鼠海马CA1区LC3II/LC3I比值、Beclin1蛋白的表达及变化情况。结果:Beclin1蛋白Western blot检测结果,与Sham组比较,VD组的Beclin1蛋白表达明显增高(P<0.05);与VD组比较,通窍活血汤高中剂量组Beclin1表达明显降低(P<0.05)。LC3蛋白Western blot检测结果:与Sham组比较,VD组的LC3II/LC3I比值均增高(P<0.05);与VD组比较,通窍活血汤高、中剂量组LC3II/LC3I比值增降低(P<0.05)。结论:通窍活血汤可减少血管性痴呆大鼠自噬相关蛋白的表达。
Objective:To investigate the effect of Tongqiao Huoxue Decoction on the expressions of autophagy related protein LC3 and Beclin-1 in hippocampus CA1 of vascular dementia rats.Methods:Fifty healthy male SD rats were randomly divided into sham operation group(Sham group),vascular dementia model group(VD group),high,middle and low dose groups of Tong Qiao Huoxue Decoction group.The rat model of vascular dementia was prepared by improved Pulsinelli four vascular blocking method.The model of Morris water maze test was used to detect the model. After 4 weeks,the LC3 II/LC3 I ratio and Beclin1 protein expressions in hippocampal CA1 area of each group were detected by Western blotting.Results:The results of Beclin1 protein was used by Western blotting.Compared with the Sham group,the expression of Beclin1 protein in VD group was significantly increased(P<0.05).Compared with the VD group,the expression of Beclin1 in the middle and high dose groups of Tongqiao Huoxue Decoction groups was significantly reduced(P<0.05).The result of LC3 protein by Western blot was compared with the sham group.The LC3 II/LC3 I ratio of VD group increased(P<0.05).Compared with the VD group,the LC3 II/LC3 I ratio in the high and middle dose groups of Tong Qiao Huoxue Decoction increased(P<0.05).Conclusion:Tongqiao Huoxue Decoction can reduce the expression of autophagy related protein in vascular dementia rats.
Objective:To investigate the effect of Tongqiao Huoxue Decoction on the expressions of autophagy related protein LC3 and Beclin-1 in hippocampus CA1 of vascular dementia rats.Methods:Fifty healthy male SD rats were randomly divided into sham operation group(Sham group),vascular dementia model group(VD group),high,middle and low dose groups of Tong Qiao Huoxue Decoction group.The rat model of vascular dementia was prepared by improved Pulsinelli four vascular blocking method.The model of Morris water maze test was used to detect the model. After 4 weeks,the LC3 II/LC3 I ratio and Beclin1 protein expressions in hippocampal CA1 area of each group were detected by Western blotting.Results:The results of Beclin1 protein was used by Western blotting.Compared with the Sham group,the expression of Beclin1 protein in VD group was significantly increased(P<0.05).Compared with the VD group,the expression of Beclin1 in the middle and high dose groups of Tongqiao Huoxue Decoction groups was significantly reduced(P<0.05).The result of LC3 protein by Western blot was compared with the sham group.The LC3 II/LC3 I ratio of VD group increased(P<0.05).Compared with the VD group,the LC3 II/LC3 I ratio in the high and middle dose groups of Tong Qiao Huoxue Decoction increased(P<0.05).Conclusion:Tongqiao Huoxue Decoction can reduce the expression of autophagy related protein in vascular dementia rats.
引文
[1] Pennisi G,Ferri R,Cantone M,et al.A review of transcranial magnetic stim-ulation in vascular dementia[J].Dement Geriatr Cogn Disord,2011,31(1):71-80.
[2] 黄玮.血管性痴呆的发病机制研究进展[J].中国医药指南,2012,10(5):62-64.
[3] 鹿俊磊,卢昌均,韦冰心,等.通窍调神法对血管性痴呆患者认知功能及事件相关电位P300的影响[J].河北中医,2015,37(3),338-341.
[4] 卢昌均,韦冰心,安红伟,等.血管性痴呆模型大鼠空间记忆障碍与海马CA1区神经元凋亡和乙酰胆碱释放有关[J].国际脑血管病杂志,2013,21(12):898-902.
[5] Altman JK,Yoon P,Katsoulidis E,et al.Regulatory effects of mammalian target of rapamycin-mediated signals in the generation of arsenic trioxide responses[J].J Biol Chem,2008,283(4):1992-2001.
[6] 晏浩,徐建军,李文林,等.MicroRNA-30a调控Beclin-1对缺氧复氧乳鼠心肌细胞的保护效应[J].中国病理生理杂志,2012,28(4):583-588.
[7] Coupe B,Ishii Y,Dietrich M O,et al.Loss of autophagy in proopiomelanocortin neurons perturbs axon growth and causes metabolic dysregulation[J].Cell Metab,2012,15(2):247-255.
[8] Quan W,Hur K Y,Lim Y,et al.Autophagy deficiency in beta-cells leads to compromised unfolded protein response and progression from obesity to diabetes in mice[J].Diabetologia,2012,55(2):392-403.
[9] 张文彦,刘金霞,刘斌,等.自噬对血管性痴呆大鼠海马CA1区生长相关蛋白-43及微管相关蛋白-2表达的影响[J].中国康复理论与实践,2016,22(7):745-749.
[10] Kang R,Zeh H J,LOTZE MT,et al.The Beclin 1 network regulates Autophagy and a poptosis[J].Cell Death Differ,2011,18(4):571-580.
[11] DJAVAHER I M M,MAIURI MC,KROEMER G.Cross talk between apoptosis and autophagy by caspase-mediated cleavage of Beclin 1[J].Oncogene,2010,29(12):1717-1719.
[12] MIZUSHIMA N.Methods for moniloring autophagy[J].Int J Biochem Cell Biol,2004,36(12):2491-2502.
[13] CHERRAS J,kulich SM,UECHI G,et al.Regulation of the autophagy protein LC3 by phosphorylation [J].J Cell Biol,2010,190(4):533-539.
[14] YAO T,YING X,ZHAO Y,et al.Vitamin D receptor activation protects against myocardial reperfusion injury through inhibition of apoptosis and modulation of autophagy[J].Antioxid Redox Signal,2015,25(22):633-650.
[15] DANIEL J,KLIONSKY Y.HAIAI A,et al.Guidelines for the use and interpretation of assays pretation of assays for monitoring autophagy in higher eukaryotes[J].Autophagy,2008,4(2):151-175.
[16] 王一超,刘斌,毛文静,等.抑制P13刚Akt/mTOR信号通路对血管性痴呆大鼠海马CA1区自噬相关蛋白表达的影响[J].中华神经医学杂志,2017,16(2)127-132.
[17] 刘斌,唐静,袁敏,等.自噬在血管性痴呆发病中的作用研究[J].临床神经病学杂志,2013,26(6):424-429.
[18] 卢昌均,周哲屹,刘国成,等.针药并用治疗血管性痴呆疗效观察[J].上海针灸杂志,2013,32(7):545-547.
[19] 国家药典委员会.中华人民共和国药典:2005年版一部[S].北京:化学工业出版社,2005:98.
[20] 汪宁,刘青云,彭代银,等.通窍活血汤对脑缺血大鼠的保护作用及机制研究[J].中国中医药信息杂志,2004,11(5):407-409.
[21] 吴秀毅,王明正.加味通窍活血汤对动物微循环的影响[J].山西医科大学学报,2002,33(6):503-505.
[2] 黄玮.血管性痴呆的发病机制研究进展[J].中国医药指南,2012,10(5):62-64.
[3] 鹿俊磊,卢昌均,韦冰心,等.通窍调神法对血管性痴呆患者认知功能及事件相关电位P300的影响[J].河北中医,2015,37(3),338-341.
[4] 卢昌均,韦冰心,安红伟,等.血管性痴呆模型大鼠空间记忆障碍与海马CA1区神经元凋亡和乙酰胆碱释放有关[J].国际脑血管病杂志,2013,21(12):898-902.
[5] Altman JK,Yoon P,Katsoulidis E,et al.Regulatory effects of mammalian target of rapamycin-mediated signals in the generation of arsenic trioxide responses[J].J Biol Chem,2008,283(4):1992-2001.
[6] 晏浩,徐建军,李文林,等.MicroRNA-30a调控Beclin-1对缺氧复氧乳鼠心肌细胞的保护效应[J].中国病理生理杂志,2012,28(4):583-588.
[7] Coupe B,Ishii Y,Dietrich M O,et al.Loss of autophagy in proopiomelanocortin neurons perturbs axon growth and causes metabolic dysregulation[J].Cell Metab,2012,15(2):247-255.
[8] Quan W,Hur K Y,Lim Y,et al.Autophagy deficiency in beta-cells leads to compromised unfolded protein response and progression from obesity to diabetes in mice[J].Diabetologia,2012,55(2):392-403.
[9] 张文彦,刘金霞,刘斌,等.自噬对血管性痴呆大鼠海马CA1区生长相关蛋白-43及微管相关蛋白-2表达的影响[J].中国康复理论与实践,2016,22(7):745-749.
[10] Kang R,Zeh H J,LOTZE MT,et al.The Beclin 1 network regulates Autophagy and a poptosis[J].Cell Death Differ,2011,18(4):571-580.
[11] DJAVAHER I M M,MAIURI MC,KROEMER G.Cross talk between apoptosis and autophagy by caspase-mediated cleavage of Beclin 1[J].Oncogene,2010,29(12):1717-1719.
[12] MIZUSHIMA N.Methods for moniloring autophagy[J].Int J Biochem Cell Biol,2004,36(12):2491-2502.
[13] CHERRAS J,kulich SM,UECHI G,et al.Regulation of the autophagy protein LC3 by phosphorylation [J].J Cell Biol,2010,190(4):533-539.
[14] YAO T,YING X,ZHAO Y,et al.Vitamin D receptor activation protects against myocardial reperfusion injury through inhibition of apoptosis and modulation of autophagy[J].Antioxid Redox Signal,2015,25(22):633-650.
[15] DANIEL J,KLIONSKY Y.HAIAI A,et al.Guidelines for the use and interpretation of assays pretation of assays for monitoring autophagy in higher eukaryotes[J].Autophagy,2008,4(2):151-175.
[16] 王一超,刘斌,毛文静,等.抑制P13刚Akt/mTOR信号通路对血管性痴呆大鼠海马CA1区自噬相关蛋白表达的影响[J].中华神经医学杂志,2017,16(2)127-132.
[17] 刘斌,唐静,袁敏,等.自噬在血管性痴呆发病中的作用研究[J].临床神经病学杂志,2013,26(6):424-429.
[18] 卢昌均,周哲屹,刘国成,等.针药并用治疗血管性痴呆疗效观察[J].上海针灸杂志,2013,32(7):545-547.
[19] 国家药典委员会.中华人民共和国药典:2005年版一部[S].北京:化学工业出版社,2005:98.
[20] 汪宁,刘青云,彭代银,等.通窍活血汤对脑缺血大鼠的保护作用及机制研究[J].中国中医药信息杂志,2004,11(5):407-409.
[21] 吴秀毅,王明正.加味通窍活血汤对动物微循环的影响[J].山西医科大学学报,2002,33(6):503-505.