右归丸对肾虚脑缺血模型大鼠行为学的影响及机制研究
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摘要
目的:通过动物实验观察肾虚对脑缺血损伤的影响及右归丸(YGP)对缺血后脑损伤的修复作用,探讨YGP是否通过调节凋亡基因的表达发挥脑保护作用。方法:选取120只无特定病原体(SPF)级雄性SD大鼠,将其分为假手术组(SOG)15只,单纯脑缺血组(SCIG)21只,肾虚脑缺血组(RACIG)21只,YGP低剂量组(YGPLG)21只,YGP高剂量组(YGPHG)21只,尼莫地平组(NpG)21只。分别采用永久性大脑中动脉阻塞(pMCAO)法和注射氢化可的松法建立脑缺血和肾虚模型,进行横木行走实验(BWT)检测运动能力;采用尼氏染色法观察神经元的状态及功能;采用免疫组化法检测脑皮质区Caspase-3的蛋白表达。各组大鼠均于术后24 h按照1次/d开始灌胃,连续灌胃14 d。SOG和SCIG灌胃等体积的生理盐水,YGPG灌胃YGP 5.13g/kg、10.26 g/kg,NpG注射Np注射液0.7 mg/kg。结果:SCIG与SOG相比,大鼠运动能力明显下降,差异有统计学意义(t=7.86,P<0.01),尼氏小体数量明显减少,Caspase-3的蛋白明显表达增加,差异有统计学意义(t=10.38,P<0.01);RACIG与SCIG相比,肾虚使缺血后脑损伤进一步加重,Caspase-3的蛋白表达明显上调,差异有统计学意义(t=13.84,P<0.01);YGP干预后,YGPHG缺血后脑损伤明显改善,Caspase-3的蛋白表达亦明显下调,YGPHG与RACIG比较,差异有统计学意义(t=6.97,P<0.01)。结论:肾虚可加重缺血后脑损伤,YGP可通过下调Caspase-3的蛋白表达发挥脑保护作用。
Objective: To observe the effect of renal asthenia on cerebral ischemia injury and the repairing effect of YouGui Pill on cerebral injury post ischemia through animal experiment, and to explore whether YouGui Pill exerted protection effect for brain by regulating the expression of apoptosis gene. Methods: 120 male SD rats of specific pathogen free(SPF) grade were randomly divided into sham operation group(SOG)(n=15), simple cerebral ischemia group(SCIG)(n=21), renal asthenia and cerebral ischemia group(RACIG)(n=21), YouGui Pill low-dose group(YGPLG)(n=21), YouGui Pill high-dose group(YGPHG)(n=21) and Nimodipine group(NpG)(n=21). Permanent middle cerebral artery occlusion(p MCAO) and hydrocortisone injection were respectively used to establish models of cerebral ischemia and renal asthenia. The motor ability was tested by beam walking test(BWT), and the state and function of neurons were observed by nissl staining, and immunohistochemistry was adopted to detect the protein expression of caspase-3 in the cerebral cortex. All rats received gavage in accordance with 1 time/d at post operative 24 h, and the period of successive gavage was 14 d post operation. The SOG and SCIG received normal saline of equal volume in gavage, and the gavage doses of YGPGs were 5.13 g/kg and10.26 g/kg, respectively, and the dose of NpG was 0.7 mg/kg. Results: Compared with SOG, the motor ability of rats of SCIG significantly decreased(t=7.86, P<0.01), and the number of nissl bodies significantly were reduced, and the protein expression of caspase-3 was significantly increased(t=10.38, P<0.01). And compared with SCIG, renal asthenia of RACIG further aggravated cerebral injury post ischemia, and the protein expression of caspase-3 was significantly upregulated(t=13.84, P<0.01). After intervention with YGP was implemented, cerebral injury post ischemia of RACIG was significantly improved, and the protein expression of caspase-3 was significantly down-regulated, and the difference of the protein expression of caspase-3 between YGPHG and RACIG was significant(t=6.97, P<0.01). Conclusion: Renal asthenia can aggravate cerebral injury post ischemia, and YGP can play a protective role for brain by down-regulating the protein expression of caspase-3.
Objective: To observe the effect of renal asthenia on cerebral ischemia injury and the repairing effect of YouGui Pill on cerebral injury post ischemia through animal experiment, and to explore whether YouGui Pill exerted protection effect for brain by regulating the expression of apoptosis gene. Methods: 120 male SD rats of specific pathogen free(SPF) grade were randomly divided into sham operation group(SOG)(n=15), simple cerebral ischemia group(SCIG)(n=21), renal asthenia and cerebral ischemia group(RACIG)(n=21), YouGui Pill low-dose group(YGPLG)(n=21), YouGui Pill high-dose group(YGPHG)(n=21) and Nimodipine group(NpG)(n=21). Permanent middle cerebral artery occlusion(p MCAO) and hydrocortisone injection were respectively used to establish models of cerebral ischemia and renal asthenia. The motor ability was tested by beam walking test(BWT), and the state and function of neurons were observed by nissl staining, and immunohistochemistry was adopted to detect the protein expression of caspase-3 in the cerebral cortex. All rats received gavage in accordance with 1 time/d at post operative 24 h, and the period of successive gavage was 14 d post operation. The SOG and SCIG received normal saline of equal volume in gavage, and the gavage doses of YGPGs were 5.13 g/kg and10.26 g/kg, respectively, and the dose of NpG was 0.7 mg/kg. Results: Compared with SOG, the motor ability of rats of SCIG significantly decreased(t=7.86, P<0.01), and the number of nissl bodies significantly were reduced, and the protein expression of caspase-3 was significantly increased(t=10.38, P<0.01). And compared with SCIG, renal asthenia of RACIG further aggravated cerebral injury post ischemia, and the protein expression of caspase-3 was significantly upregulated(t=13.84, P<0.01). After intervention with YGP was implemented, cerebral injury post ischemia of RACIG was significantly improved, and the protein expression of caspase-3 was significantly down-regulated, and the difference of the protein expression of caspase-3 between YGPHG and RACIG was significant(t=6.97, P<0.01). Conclusion: Renal asthenia can aggravate cerebral injury post ischemia, and YGP can play a protective role for brain by down-regulating the protein expression of caspase-3.
引文
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[15]苗光新,李蒙蒙,吴晓光.山楂叶总黄酮对脑缺血大鼠Caspase-3蛋白及炎症因子表达的影响[J].中国老年学杂志,2019,39(10):2484-2490.
[2]李菲,张洋洋,蔡锐,等.淫羊藿苷通过上调VEGF信号通路促进脑低灌注大鼠的血管发生[J].遵义医学院学报,2018,41(5):533-538.
[3]肖坤.地黄饮子汤对大鼠脑缺血再灌注损伤的影响[J].中西医结合心脑血管病,2018,16(11):1502-1504.
[4]赵英华,曾维.六味地黄丸加减治疗脑梗死恢复期的临床效果[J].黑龙江医药,2018,31(1):85-86.
[5]徐伟,王丽娜,王键,等.益气活血方和补肾生髓方对脑缺血再灌注大鼠海马和额顶叶皮质Jagged1、Notch4和Hes1蛋白表达的影响[J].云南中医学院学报,2017,40(6):1-13.
[6]张雨薇,刘宏,张博,等.补肾活血方对PMCAO模型大鼠行为学及脑梗死体积的影响[J].中医药信息,2016,33(1):8-12.
[7]方丽媛,王平,谭爱华,等.淫羊藿苷和固本方对肾虚型老年痴呆大鼠行为及病理学的作用[J].中华中医药杂志,2017,32(12):5322-5326.
[8]詹正烜,郑乃熙,张英杰,等.肾阴虚、肾阳虚模型大鼠股骨皮质骨差异蛋白质表达的分析[J].中国中西医结合杂志,2017,37(10):1242-1247.
[9]张小琴,侯丽颖,强国芳,等.片仔癀对局灶性脑缺血/再灌注大鼠皮质中NMDAR1和GluR2表达的影响[J].中国药理学通报,2019,35(2):288-292.
[10]刘宏,刘学伟,仲丽丽,等.右归丸改善睾丸摘除雄性MCAO模型大鼠运动能力的实验研究[J].黑龙江医学,2017,37(6):670-673.
[11]张博,叶阳,刘学伟,等.电针改善去势雌性大鼠记忆障碍的机制研究[J].湖南中药大学学报,2017,37(6):670-673.
[12]郎丰山,黄云霞,薛云,等.脑缺血再灌注损伤病理生理机制研究进展[J].食品与药品,2017,20(4):312-316.
[13]Iadecola C,Anrather A.Stroke research at acrossroad:the brain for directions[J].Neurosci,2011,14(11):1363-1368.
[14]彭智远,刘旺华,曹雯.脑缺血再灌注损伤细胞凋亡机制的研究进展[J].中华中医药学刊,2017,53(8):1975-1961.
[15]苗光新,李蒙蒙,吴晓光.山楂叶总黄酮对脑缺血大鼠Caspase-3蛋白及炎症因子表达的影响[J].中国老年学杂志,2019,39(10):2484-2490.