人参皂苷Rg1对大鼠脊髓缺血再灌注损伤中survivin蛋白表达及细胞凋亡的影响
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摘要
目的观察人参皂苷Rg1预处理对于大鼠脊髓缺血再灌注损伤后的survivin蛋白表达及细胞凋亡的影响,探讨人参皂苷Rg1对脊髓缺血再灌注损伤后大鼠运动功能恢复的可能机制。方法选取成年健康的SD大鼠120只,构建大鼠脊髓缺血再灌注损伤模型,随机分为假手术组、缺血组、缺血再灌注组及药物组。采用后肢神经运动功能(BBB)评价大鼠后肢神经运动功能;免疫组化法观察survivin蛋白及凋亡诱导因子(apoptosis-inducing factor, AIF)表达;Western blot及RT-PCR方法分析各组survivin及Caspase-9的表达和活性。结果人参皂苷Rg1的干预可以使大鼠后肢神经运动功能评分升高,AIF阳性细胞数减少、survivin蛋白阳性细胞数增多、survivin及Caspase-9的表达减少、神经细胞凋亡减少。结论人参皂苷Rg1可能通过促进survivin蛋白的表达来抑制Caspase-9表达以及降低胞质AIF的水平介导大鼠脊髓缺血再灌注损伤造成的细胞凋亡。
Objective To observe the effects of ginsenoside Rg1 pretreatment on the expression of survivin protein and apoptosis after spinal cord ischemia-reperfusion injury(SCII) in rats so as to explore the possible mechanism of ginsenoside Rg1 on motor function recovery after SCII in rats. Methods We selected 120 adult healthy SD rats to construct the model of SCII and randomly divided them into four groups: sham operation group, ischemia group, ischemia-reperfusion group, and drug group. Basso Beattie and Bresnahan score(BBB score) was used to evaluate the motor function of the hind limbs of the rats. The expressions of survivin protein and apoptosis-inducing factor(AIF) was observed by immunohistochemistry. The expression and activity of survivin protein and Caspase-9 in each group were observed and analyzed by Western blot and RT-PCR. Results The intervention of ginsenoside Rg1 could increase the score of the motor function of the rat hind limbs. It could decrease the number of AIF positive cells, but increase the number of survivin protein positive cells. Ginsenoside Rg1 could decrease the expressions of survivin and Caspase-9, and decrease the apoptosis of nerve cells in SCII. Conclusion Ginsenoside Rg1 could inhibit the expression of Caspase-9 by promoting the expression of survivin protein and decrease the apoptosis of rat SCII induced by the level of cytoplasmic AIF.
Objective To observe the effects of ginsenoside Rg1 pretreatment on the expression of survivin protein and apoptosis after spinal cord ischemia-reperfusion injury(SCII) in rats so as to explore the possible mechanism of ginsenoside Rg1 on motor function recovery after SCII in rats. Methods We selected 120 adult healthy SD rats to construct the model of SCII and randomly divided them into four groups: sham operation group, ischemia group, ischemia-reperfusion group, and drug group. Basso Beattie and Bresnahan score(BBB score) was used to evaluate the motor function of the hind limbs of the rats. The expressions of survivin protein and apoptosis-inducing factor(AIF) was observed by immunohistochemistry. The expression and activity of survivin protein and Caspase-9 in each group were observed and analyzed by Western blot and RT-PCR. Results The intervention of ginsenoside Rg1 could increase the score of the motor function of the rat hind limbs. It could decrease the number of AIF positive cells, but increase the number of survivin protein positive cells. Ginsenoside Rg1 could decrease the expressions of survivin and Caspase-9, and decrease the apoptosis of nerve cells in SCII. Conclusion Ginsenoside Rg1 could inhibit the expression of Caspase-9 by promoting the expression of survivin protein and decrease the apoptosis of rat SCII induced by the level of cytoplasmic AIF.
引文
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[21] YANG X,CHENG J,GAO Y,et al.Downregulation of Iduna is associated with AIF nuclear translocation in neonatal brain after hypoxia-ischemia[J].Neuroscience,2017,346:74-80.
[22] 程斌,王鑫,王磊,等.兔脊髓缺血再灌注损伤后线粒体凋亡途径的变化及灯盏细辛预处理的干预作用[J].中西医结合学报,2009,7(9):842-847.
[2] NAKKA VP,GUSAIN A,MEHTA SL,et al.Molecular mechanisms of apoptosis in cerebral ischemia:Multiple neuroprotective opportunities[J].Mol Neurobiol,2008,37(1):7-38.
[3] YU ZQ,WEI YH,XIN G,et al.Effects of transplantation with marrow-derived mesenchymal stem cells modified with survivin on renal ischemia-reperfusion injury in mice[J].Yonsei Med J,2014,55(4):1130-1137.
[4] SCHEER A,KNAUER S K,VERHAEGH R.Survivin expression pattern in the intestine of normoxic and ischemic rats[J].BMC Gastroenterol,2017,17(1):76.
[5] LEE S,RHEE D K.Effects of ginseng on stress-related depression,anxiety,and the hypothalamic-pituitary-adrenal axis[J].J Ginseng Res,2017,41(4):589-594.
[6] YU S,ZHOU X,LI F,et al.Microbial transformation of ginsenoside Rb1,Re and Rg1 and its contribution to the improved anti-inflammatory activity of ginseng[J].Sci Rep,2017,7(1):138.
[7] 李亮,邓文祥,何军锋,等.人参皂苷Rg1对局灶性脑缺血再灌注大鼠的神经保护作用[J].神经损伤与功能重建,2016,11(2):95-98.
[8] SONG RB,BASSO DM,DA CR,et al.Adaptation of the Basso-Beattie-Bresnahan locomotor rating scale for use in a clinical model of spinal cord injury in dogs[J].J Neurosci Methods,2016,268:117-124.
[9] 夏炳江,肖鲁伟.脊髓慢性压迫减压后缺血再灌注损伤研究进展[J].中华中医药学刊,2017,35(4):998-1001.
[10] ROGERS C,FERNANDES-ALNEMRI T,MAYES L,et al.Cleavage of DFNA5 by Caspase-3 during apoptosis mediates progression to secondary necrotic/pyroptotic cell death[J].Nat Commun,2017,8:14128.
[11] LIANG S,SUN K,WANG Y,et al.Role of Cyt-C/caspases-9,3,Bax/Bcl-2 and the FAS death receptor pathway in apoptosis induced by zinc oxide nanoparticles in human aortic endothelial cells and the protective effect by alpha-lipoic acid[J].Chem Biol Interact,2016,258:40-51.
[12] ZHANG Q,LIU J,CHEN S,et al.Caspase-12 is involved in stretch-induced apoptosis mediated endoplasmic reticulum stress[J].Apoptosis,2016,21(4):432-442.
[13] SAKURAI M,NAGATA T,ABE K,et al.Survival and death-promoting events after transient spinal cord ischemia in rabbits:induction of Akt and caspase3 in motor neurons[J].J Thorac Cardiovasc Surg,2003,125(2):370-377.
[14] TANG L,SHEN H,LI X,et al.MiR-125a-5p decreases after long non-coding RNA HOTAIR knockdown to promote cancer cell apoptosis by releasing caspase 2[J].Cell Death Dis,2016,7:e2137.
[15] ZHAO YM,WU KN,WANG YJ,et al.Synergistic cytotoxic effects of rapamycin and idarubicin on human acute T-cell lymphoblastic leukemia Jurkat cells[J].Zhejiang Da Xue Xue Bao (Yi Xue Ban),2011,40(5):482-488.
[16] LEIST M,SINGLE B,CASTOLDI AF,et al.Intracellular adenosine triphosphate (ATP) concentration:A switch in the decision between apoptosis and necrosis[J].J Exp Med,1997,185(8):1481-1486.
[17] KIM HE,DU F,FANG M,et al.Formation of apoptosome is initiated by cytochrome c-induced dATP hydrolysis and subsequent nucleotide exchange on Apaf-1[J].Proc Natl Acad Sci U S A,2005,102(49):17545-17550.
[18] GARG H,SURI P,GUPTA JC,et al.Survivin:A unique target for tumor therapy[J].Cancer Cell Int,2016,16:49.
[19] VOGL T J,OPPERMANN E,QIAN J,et al.Transarterial chemoembolization of hepatocellular carcinoma in a rat model:The effect of additional injection of survivin siRNA to the treatment protocol[J].BMC Cancer,2016,16:325.
[20] MIRAMAR MD,COSTANTINI P,RAVAGNAN L,et al.NADH oxidase activity of mitochondrial apoptosis-inducing factor[J].J Biol Chem,2001,276(19):16391-16398.
[21] YANG X,CHENG J,GAO Y,et al.Downregulation of Iduna is associated with AIF nuclear translocation in neonatal brain after hypoxia-ischemia[J].Neuroscience,2017,346:74-80.
[22] 程斌,王鑫,王磊,等.兔脊髓缺血再灌注损伤后线粒体凋亡途径的变化及灯盏细辛预处理的干预作用[J].中西医结合学报,2009,7(9):842-847.
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