复方脑肽节苷脂对脑缺血再灌注损伤模型大鼠的保护作用
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摘要
目的探讨复方脑肽节苷脂对大鼠脑缺血再灌注损伤模型大鼠的保护作用。方法将120只SD大鼠随机分为假手术组(等体积0. 9%氯化钠注射液)、模型组(等体积0. 9%氯化钠注射液)、金纳多组(50 mg/kg)与复方脑肽节苷脂低、中、高剂量组(2,4,8 mg/kg),各20只。模型组、金纳多组、复方脑肽节苷脂各剂量组大鼠予缺血再灌注以建立模型,假手术组不阻塞血流。建模成功后各组大鼠腹腔注射相应药物,每天1次,持续14 d。给药7 d及14 d时进行神经运动功能评分;给药14 d后进行贴纸去除及平衡木行走试验,采用逆转录定量聚合酶链反应(PCR)法,酶联免疫吸附(ELISA)法和免疫组化法分别检测大鼠缺血脑组织Beclin1 mRNA,Parkin mRNA,PINK1mRNA及蛋白表达水平。结果与假手术组比较,模型组大鼠神经功能缺损评分显著升高,双侧贴纸去除时间、平衡木过杆时间均显著延长,Parkin mRNA和蛋白表达水平均显著升高(P <0. 05),Beclin1 mRNA,PINK1 mRNA和蛋白表达水平均显著降低(P <0. 05);与模型组比较,金纳多组及复方脑肽节苷脂低、中、高剂量组神经功能缺损评分均显著降低,双侧贴纸去除时间、平衡木过杆时间均显著缩短,Parkin mRNA和蛋白表达水平均显著降低,Beclin1 mRNA,PINK1 mRNA和蛋白表达水平均显著升高(P <0. 05)。结论复方脑肽节苷脂对脑缺血再灌注损伤模型大鼠具有较好的保护作用,能明显改善其神经、感觉及运动功能,其机制与其促进Beclin1 mRNA,PINK1mRNA及蛋白表达,抑制Parkin表达,从而抑制神经细胞凋亡有关。
Objective To investigate the protective effect of Compound Porcine Cerebroside and Ganglioside Injection( CPCGI) on cerebral ischemical reperfusion injury in model rats. Methods Totally 120 SD rats were randomly divided into 6 groups: the sham operation group( equal volume of 0. 9% Sodium Chloride Injection),model group( equal volume of 0. 9% Sodium Chloride Injection),ginkgo group( 50 mg/kg) and CPCGI low,medium,high dose groups( 2,4,8 mg/kg),20 rats in each group. Rats in the model group,ginkgo group and CPCGI groups were subjected to ischemical reperfusion to establish the models,while the blood flow of rats in the sham operation group was not obstructed. After 1 d of successful modeling, rats in each group were intraperitoneally injected with corresponding drugs once a day for 14 consecutive days. 7 and 14 d after administration,the neuromotor function scores were scored. After 14 d of administration,sticker removal and balance beam walking test were carried out. The expression levels of Beclin1 mRNA, Parkin mRNA,PINK1 mRNA and protein in brain tissue of rats were detected by reverse transcription polymerase chain reaction( PCR), enzyme-linked immunosorbent assay( ELISA) and immunohistochemistry respectively. Results Compared with the sham operation group, the neurological deficit score increased significantly,the removal time of bilateral stickers and the time of crossing the balance beam prolonged significantly, the expression levels of Parkin mRNA and protein increased significantly( P < 0. 05),and the expression levels of Beclin1 mRNA,PINK1 mRNA and protein decreased significantly in the model group( P < 0. 05). Compared with the model group, the neurological deficit scores decreased significantly,the removal time of bilateral stickers and the time of crossing the balance beam shortened significantly,the expression levels of Parkin mRNA and protein decreased significantly,the expression levels of Beclin1 mRNA,PINK1 mRNA and protein increased significantly in the ginkgo group and the CPCGI low,medium,high dose groups( P < 0. 05). Conclusion CPCGI has good protective effect on cerebral ischemical reperfusion injury model rats, and can significantly improve their neurological, sensory and motor functions. Its mechanism is related to promoting the expression of Beclin1 mRNA, PINK1 mRNA and protein, inhibiting the expression of Parkin and inhibiting the apoptosis of nerve cells.
Objective To investigate the protective effect of Compound Porcine Cerebroside and Ganglioside Injection( CPCGI) on cerebral ischemical reperfusion injury in model rats. Methods Totally 120 SD rats were randomly divided into 6 groups: the sham operation group( equal volume of 0. 9% Sodium Chloride Injection),model group( equal volume of 0. 9% Sodium Chloride Injection),ginkgo group( 50 mg/kg) and CPCGI low,medium,high dose groups( 2,4,8 mg/kg),20 rats in each group. Rats in the model group,ginkgo group and CPCGI groups were subjected to ischemical reperfusion to establish the models,while the blood flow of rats in the sham operation group was not obstructed. After 1 d of successful modeling, rats in each group were intraperitoneally injected with corresponding drugs once a day for 14 consecutive days. 7 and 14 d after administration,the neuromotor function scores were scored. After 14 d of administration,sticker removal and balance beam walking test were carried out. The expression levels of Beclin1 mRNA, Parkin mRNA,PINK1 mRNA and protein in brain tissue of rats were detected by reverse transcription polymerase chain reaction( PCR), enzyme-linked immunosorbent assay( ELISA) and immunohistochemistry respectively. Results Compared with the sham operation group, the neurological deficit score increased significantly,the removal time of bilateral stickers and the time of crossing the balance beam prolonged significantly, the expression levels of Parkin mRNA and protein increased significantly( P < 0. 05),and the expression levels of Beclin1 mRNA,PINK1 mRNA and protein decreased significantly in the model group( P < 0. 05). Compared with the model group, the neurological deficit scores decreased significantly,the removal time of bilateral stickers and the time of crossing the balance beam shortened significantly,the expression levels of Parkin mRNA and protein decreased significantly,the expression levels of Beclin1 mRNA,PINK1 mRNA and protein increased significantly in the ginkgo group and the CPCGI low,medium,high dose groups( P < 0. 05). Conclusion CPCGI has good protective effect on cerebral ischemical reperfusion injury model rats, and can significantly improve their neurological, sensory and motor functions. Its mechanism is related to promoting the expression of Beclin1 mRNA, PINK1 mRNA and protein, inhibiting the expression of Parkin and inhibiting the apoptosis of nerve cells.
引文
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[11]李军汉,苏全生,孙君志,等.运动和饮食调整对内质网应激介导非酒精性脂肪肝大鼠模型肝细胞凋亡的影响[J].中国运动医学杂志,2017,36(1):36-43.
[12] XIAO B,GOH JY,XIAO L,et al. Reactive oxygen species trigger Parkin/PINK1 pathway-dependent mitophagy by inducing mitochondrial recruitment of Parkin[J]. Journal of Biological Chemistry,2017,292(40):97-98.
[13] REN X,LUO P,ZHOU H,et al. Resveratrol Ameliorates Mitochondrial Elongation via Drp1/Parkin/PINK1 Signaling in Senescent-Like Cardiomyocytes[J]. Oxidative Medicine and Cellular Longevity,2017,2017:4195353.
[14] XIAO B,DENG X,LIM GGY,et al. Superoxide drives progression of Parkin/PINK1-dependent mitophagy following translocation of Parkin to mitochondria[J]. Cell Death&Disease,2017,8(10):97-99.
[15] MIAO T,YU C,DAN T,et al. Beclin1 antagonizes LAPTM4Bmediated EGFR overactivation in gastric cancer cells[J]. Gene,2017,626(2):48-50.
[16]赵丹,胡东懿,岑敏,等.脑苷肌肽注射液与复方脑肽节苷脂注射液治疗新生儿缺氧缺血性脑病的临床疗效比较和成本分析[J].实用心脑肺血管病杂志,2018,6(2):76-78.
[2] DU X. Inhibition of cerebral ischemia/reperfusion injuryinduced apoptosis:nicotiflorin and JAK2/STAT3 pathway[J]. Neural Regeneration Research,2017,12(1):96-102.
[3] LI Y,LIU S. The Effect of Dexmedetomidine on Oxidative Stress Response Following Cerebral Ischemia-Reperfusion in Rats and the Expression of Intracellular Adhesion Molecule-1(ICAM-1)and S100B[J]. Medical Science Monitor International Medical Journal of Experimental&Clinical Research,2017,23:867-873.
[4] GONG J,WANG XZ. Signal pathway and regulation mechanism of endoplasmic reticulum stress[J]. Journal of Zhejiang UniversityScience B(Biomedicine&Biotechnology),2017,18(1):1-14.
[5]王沛坚,王芳,万进东,等.内质网应激及线粒体功能障碍在糖尿病血管内皮损伤中作用的研究进展[J].现代生物医学进展,2017,17(2):389-392.
[6] MCWILLIAMS TG,MUQIT MM. PINK1 and Parkin:emerging themes in mitochondrial homeostasis[J]. Curr Opin Cell Biol,2017,45:83-91.
[7]黄伟毅,关良劲.亚低温联合复方脑肽节苷脂注射液治疗重症颅脑损伤的临床疗效及生活质量研究[J].脑与神经疾病杂志,2018,26(2):100-104.
[8]谭杰,李承晏,李涛,等.局灶性脑梗死模型大鼠相对体质量、Bederson评分与梗死类型的鉴别[J].中国组织工程研究,2011,15(37):6875-6878.
[9] ALTUMBABIC M,PEELING J,DEL BIGIO MR. Intracerebral hemor-rhage in the rat:effects of hematoma aspiration[J].Stroke,1998,29(9):1917-1923.
[10]许斌,李岚,宋启斌.内质网应激的致癌性和免疫抑制效应[J].肿瘤学杂志,2018,24(5):492-498.
[11]李军汉,苏全生,孙君志,等.运动和饮食调整对内质网应激介导非酒精性脂肪肝大鼠模型肝细胞凋亡的影响[J].中国运动医学杂志,2017,36(1):36-43.
[12] XIAO B,GOH JY,XIAO L,et al. Reactive oxygen species trigger Parkin/PINK1 pathway-dependent mitophagy by inducing mitochondrial recruitment of Parkin[J]. Journal of Biological Chemistry,2017,292(40):97-98.
[13] REN X,LUO P,ZHOU H,et al. Resveratrol Ameliorates Mitochondrial Elongation via Drp1/Parkin/PINK1 Signaling in Senescent-Like Cardiomyocytes[J]. Oxidative Medicine and Cellular Longevity,2017,2017:4195353.
[14] XIAO B,DENG X,LIM GGY,et al. Superoxide drives progression of Parkin/PINK1-dependent mitophagy following translocation of Parkin to mitochondria[J]. Cell Death&Disease,2017,8(10):97-99.
[15] MIAO T,YU C,DAN T,et al. Beclin1 antagonizes LAPTM4Bmediated EGFR overactivation in gastric cancer cells[J]. Gene,2017,626(2):48-50.
[16]赵丹,胡东懿,岑敏,等.脑苷肌肽注射液与复方脑肽节苷脂注射液治疗新生儿缺氧缺血性脑病的临床疗效比较和成本分析[J].实用心脑肺血管病杂志,2018,6(2):76-78.