补阳还五汤对脑缺血再灌注损伤大鼠PTEN mRNA的影响
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摘要
目的:探讨补阳还五汤对脑缺血再灌注大鼠第10号染色体缺失的磷酸酶和张力蛋白同源物基因(PTEN)mRNA表达与血小板活化的影响。方法:将无特定病原体(SPF)级SD大鼠60只随机分为假手术组、模型组、氢氯吡格雷组、补阳还五汤高、低剂量组,每组12只。氢氯吡格雷组ig给予氯吡格雷6.8 mg·kg-1·d-1,补阳还五汤高、低剂量组分别ig给予补阳还五汤26,6.5 g·kg-1·d-1,其余各组ig给予生理盐水,连续给药14 d后,采用大脑中动脉线栓法复制大鼠急性局灶性脑缺血再灌注模型,并在造模前2 h给予相应ig处理,造模60 min后拔除栓线再灌注12 h后收集相应标本保存,测定各组大鼠血浆P-选择素(CD62P)含量与海马组织PTEN mRNA表达情况。结果:与假手术组比较,模型组神经行为评分显著增加,与模型组比较,各用药组的神经行为评分显著降低;与假手术组比较,模型组血浆中CD62P表达显著升高(P<0.05);与模型组比较,补阳还五汤组与氢氯吡格雷组均能显著抑制血浆中CD62P表达(P<0.05),但补阳还五汤组与氢氯匹格雷组之间抑制作用无显著性差异;与假手术组比较,模型组PTEN mRNA表达显著增加;与模型组比较,各用药组的PTEN mRNA表达显著降低(P<0.05),以补阳还五汤高剂量组尤为明显。结论:补阳还五汤对急性脑缺血再灌注损伤模型大鼠的保护机制可能与下调PTEN基因过表达及抑制CD62P表达有关。
Objective: To investigate the effect of Buyang Huanwu Tang( BYHWT) on the expression of phosphatase and tensin homolog deleted on chromosome ten( PTEN) mRNA and platelet activation in rat models with cerebral ischemia-reperfusion injury. Method: Sixty specific pathogen free( SPF) Sprague-Dawley rats( weighing 250-300 g) were randomly divided into 5 groups as following: sham-operated group,clopidogrel group,high-dose BYHWT group,low-dose BYHWT group,and model group,12 rats in each group. The high-dose BYHWT group and the low-dose BYHWT group were ig administered with 26,6. 5 g ·kg- 1·d- 1BYHWT respectively while the clopidogrel group was ig administered with 6. 8 mg·kg- 1·d- 1. Another two groups were ig administered with normal saline. After treatment of the animals for 14 days, middle cerebral artery occlusion method was used to establish models of transient focal cerebral ischemia and reperfusion,and corresponding ig treatment was provided 2 h before modeling. The plugging wire was removed after modeling for 60 min and reperfusion for 12 h before samples collection for determination of the content of P-selection( CD62P) in plasma and expression of PTEN mRNA in hippocampus. Result: The neurological score in model group was significantly higher than that in sham-operated group,and the neurological scores in various treatment groups were significantly lower than that in model group. The expression of CD62 P in the model group was significantly higher than that in sham-operated group( P < 0. 05). BYHWT groups and clopidogrel group can significantly inhibit the expression of CD62P( P < 0. 05),but without significant difference between BYHWT groups and clopidogrel group. PTEN mRNA expression in model group was significantly higher than that in sham-operated group. PTEN mRNA expression in various treatment groups were significantly lower than that in model group( P < 0. 05),mostly significant in BYHWT high dose group. Conclusion: The neuroprotective mechanism of BYHWT on acute cerebral ischemia model may relate with down-regulating the expression of PTEN gene over-expression and inhibiting expression of CD62 P.
Objective: To investigate the effect of Buyang Huanwu Tang( BYHWT) on the expression of phosphatase and tensin homolog deleted on chromosome ten( PTEN) mRNA and platelet activation in rat models with cerebral ischemia-reperfusion injury. Method: Sixty specific pathogen free( SPF) Sprague-Dawley rats( weighing 250-300 g) were randomly divided into 5 groups as following: sham-operated group,clopidogrel group,high-dose BYHWT group,low-dose BYHWT group,and model group,12 rats in each group. The high-dose BYHWT group and the low-dose BYHWT group were ig administered with 26,6. 5 g ·kg- 1·d- 1BYHWT respectively while the clopidogrel group was ig administered with 6. 8 mg·kg- 1·d- 1. Another two groups were ig administered with normal saline. After treatment of the animals for 14 days, middle cerebral artery occlusion method was used to establish models of transient focal cerebral ischemia and reperfusion,and corresponding ig treatment was provided 2 h before modeling. The plugging wire was removed after modeling for 60 min and reperfusion for 12 h before samples collection for determination of the content of P-selection( CD62P) in plasma and expression of PTEN mRNA in hippocampus. Result: The neurological score in model group was significantly higher than that in sham-operated group,and the neurological scores in various treatment groups were significantly lower than that in model group. The expression of CD62 P in the model group was significantly higher than that in sham-operated group( P < 0. 05). BYHWT groups and clopidogrel group can significantly inhibit the expression of CD62P( P < 0. 05),but without significant difference between BYHWT groups and clopidogrel group. PTEN mRNA expression in model group was significantly higher than that in sham-operated group. PTEN mRNA expression in various treatment groups were significantly lower than that in model group( P < 0. 05),mostly significant in BYHWT high dose group. Conclusion: The neuroprotective mechanism of BYHWT on acute cerebral ischemia model may relate with down-regulating the expression of PTEN gene over-expression and inhibiting expression of CD62 P.
引文
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[15]尹天雷,蔡光先,李勇敏,等.补阳还五汤对脑缺血模型大鼠脑组织NGF及PI3K/AKT信号转导途径的影响[J].中国中医药科技,2008,15(1):24-25.
[16]王江友,李浪.PTEN在心肌缺血/再灌注损伤中的研究进展[J].中国老年学杂志,2013,33(17):4367-4369.
[17]Laurent P A,Severin S,Gratacap M P,et al.Class I PI 3-kinases signaling in platelet activation and thrombosis:PDK1/Akt/GSK3 axis and impact of PTEN and SHIP1[J].Adv Biol Regul,2014,54:162-174.
[18]苏强,李浪.PTEN与炎症关系的研究进展[J].重庆医学,2014,43(4):494-497.
[19]曲娟,朱晓东,胡进.丹红注射液对短暂性脑缺血发作患者CD62P.CD63的影响[J].浙江中医杂志,2014,49(9):636-637.
[2]Chen Z Y,Cao L,Wang L M,et al.Development of neurotrophic molecules for treatment of neurodegeneration[J].Curr Protein Pept Sci,2001,2(3):261-276.
[3]张继平,宫丽,李齐欢,等.补阳还五汤家兔含药血清对家兔血小板PAF受体活性的影响[J].中国中医基础医学杂志,2010,16(8):688-690.
[4]高旭光.2011版美国心脏协会/美国卒中学会卒中预防指南解读[J].中国医学前沿杂志:电子版,2011,3(3):78-80.
[5]Schattner M.Platelets and galectins[J].Ann Transl Med,2014,2(9):85.
[6]Weng Z,Li D,Zhang L,et al.PTEN regulates collagen-induced platelet activation[J].Blood,2010,116(14):2579-2581.
[7]蔡俊,张继平,姚晖,等.补阳还五汤对急性脑缺血再灌注大鼠脑组织AKT和p-AKT蛋白表达的影响[J].中国实验方剂学杂志,2015,21(6):122-126
[8]Longa E Z,Weinstein P R,Carlson S,et al.Reversible middle cerebral artery occlusion without craniectomy in rats[J].Stroke,1989,20(1):84-91.
[9]周赛男,蔺晓源,易健,等.补阳还五汤对脑缺血大鼠神经功能及细胞形态的影响[J].中国实验方剂学杂志,2013,19(2):251-254.
[10]霍勇,陆菊明.国内外指南推荐阿司匹林用于心脑血管疾病一级预防[J].中华内科杂志,2013,52(10):882-885.
[11]Grundmann K,Jaschonek K,Kleine B,et al.Aspirin non-responder status in patients with recurrent cerebral ischemic attacks[J].J Neurol,2003,250(1):63-66.
[12]Gurbel P A,Bliden K P,Hiatt B L,et al.Clopidogrel for coronary stenting:response variability,drug resistance,and the effect of pretreatment platelet reactivity[J].Circulation,2003,107(23):2908-2913.
[13]Mu Q,Liu P,Hu X,et al.Neuroprotective effects of Buyang Huanwu decoction on cerebral ischemia-induced neuronal damage[J].Neural Regen Res,2014,9(17):1621-1627.
[14]Fan X H,Shi W Z,Cheng Y X,et al.Effects of Buyang Huanwu Decoction on antioxidant and drugmetabolizing enzymes in rat liver[J].Chin J Nat Med,2014,12(6):449-454.
[15]尹天雷,蔡光先,李勇敏,等.补阳还五汤对脑缺血模型大鼠脑组织NGF及PI3K/AKT信号转导途径的影响[J].中国中医药科技,2008,15(1):24-25.
[16]王江友,李浪.PTEN在心肌缺血/再灌注损伤中的研究进展[J].中国老年学杂志,2013,33(17):4367-4369.
[17]Laurent P A,Severin S,Gratacap M P,et al.Class I PI 3-kinases signaling in platelet activation and thrombosis:PDK1/Akt/GSK3 axis and impact of PTEN and SHIP1[J].Adv Biol Regul,2014,54:162-174.
[18]苏强,李浪.PTEN与炎症关系的研究进展[J].重庆医学,2014,43(4):494-497.
[19]曲娟,朱晓东,胡进.丹红注射液对短暂性脑缺血发作患者CD62P.CD63的影响[J].浙江中医杂志,2014,49(9):636-637.