背景:临床药物单独使用对脊髓损伤特定病理变化具有较好地阻断作用,但单一用药往往存在较为复杂的不良反应,如雌激素可以缓解氧化损伤导致的细胞凋亡,但同时会激活体内的炎症反应.在此大背景下,联合协同用药成为临床创新发展的趋势,其联合用药机制也迫切需要阐明.目的:探究地塞米松联合雌激素治疗对脊髓钝挫伤大鼠白细胞介素6、Caspase3和Bcl-2表达的影响.方法:雌性4周龄SD大鼠48只,随机分为假手术组、生理盐水组、地塞米松组、联合治疗组,每组12只.假手术组显露脊髓后直接缝合,其他3组采用改良Allen's法建立脊髓损伤模型,术后30 min和1,2,3,4,5d时假手术组和生理盐水组大鼠注射生理盐水(0.2 mL/kg),地塞米松组大鼠注射地塞米松(0.2 mg/kg),联合治疗组大鼠先后注射雌二醇(0.04 mg/kg)、地塞米松(0.2 mg/kg),注射部位均为大鼠前肢大腿外侧肌.术后14d内采用BBB评分观察大鼠后肢运动功能恢复情况,术后3,7,21d采用免疫组织化学检测脊髓组织中白细胞介素6、Bcl-2和Caspase3的表达.结果 与结论:①联合治疗组运动功能恢复开始时间较地塞米松组和生理盐水组更早,在整个恢复时期,联合治疗组的BBB评分显著高于地塞米松组(P＜0.05);②与地塞米松组相比,联合治疗组白细胞介素6和Casepase3表达显著降低、Bcl-2表达明显升高(P＜0.05);③结果表明,联合治疗能有效减少脊髓损伤后促凋亡因子Caspase3和促炎因子白细胞介素6的表达,增加抗凋亡因子Bcl-2的表达,通过抑制凋亡和炎症促进脊髓损伤后功能恢复.

目的:探究神经生物电刺激对原发性痛经(Primary dysmenorrhea,PD)大鼠P物质(Substance P,SP)的表达变化的影响.方法:将SD大鼠随机分为非模型对照组、模型对照组和实验组3组.实验组给予股部皮下注射已烯雌酚,每天一次,连续10d,第1天0.8 mg·只-1,第2-9天0.4 mg·只-1,第10天0.8 mg·只-1;第11天腹腔注射缩宫素2U·只-1;对实验组大鼠进行神经生物电刺激干预,并在1h、3h和7h后采用扭体评分进行运动评价.模型对照组只注射与实验组相同剂量的已烯雌酚与缩宫素,不进行神经生物电刺激干预,其余操作不变.非模型对照组不注射已烯雌酚与缩宫素,不进行神经生物电刺激干预,其余操作不变.免疫组织化学技术检测各组大鼠脊髓后角中SP的定位及表达变化,采用免疫荧光技术探究SP与脑源性神经营养因子(Brain derived neurotrophic factor,BDNF)在脊髓后角的共定位情况.结果:扭体评分显示实验组大鼠运动增加幅度明显小于模型对照组(P＜0.05).SP主要分布于脊髓后角Ⅱ板层,所有时间点实验组SP表达均低于模型对照组,在3h表达差异最大(P＜0.05).免疫荧光双标技术显示SP和BDNF在脊髓后角共表达.结论:神经生物电刺激能抑制SP的表达,影响原发性痛经,其机制可能与SP及BDNF共同作用有关.

目的 观察突触活性带蛋白1 (Complexin-1,CPLX-1)在大鼠脊髓损伤后的表达变化.方法 采用Allen's法制备SD大鼠脊髓钝挫伤模型,BBB评分评估大鼠的运动功能,免疫组织化学技术观察CPLX-1在脊髓中分布,实时定量PCR(qRT-PCR)检检测CPLX-1基因在损伤后的表达变化.结果 脊髓损伤后,大鼠后肢运动功能明显障碍,BBB评分于损伤后第1d最低,术后7d才有所恢复(p＜0.05).免疫组织化学结果显示,CPLX-1在脊髓前角运动神经元和神经胶质细胞中表达.此外,脊髓损伤后CPLX-1在神经元中表达逐渐降低,至术后第3d时最低,后随时间逐渐增加,但在神经胶质细胞中表达逐渐增加,至术后第3d时最高,后随时间逐渐降低;qRT-PCR结果显示脊髓损伤后CPLX-1的基因表达急剧下降,至术后3d达到最低,随后逐渐上升,但一直到术后28d仍低于正常水平(p＜0.05).结论 CPLX-1在脊髓运动神经元和神经胶质细胞中表达,在脊髓损伤后其基因和蛋白的表达量下降,可能在大鼠脊髓损伤后运动功能的恢复中起重要作用.

Objective To explore the change of apoptosis factor Caspase-3 and Bcl-2 in the injured segment of rat with spinal cord injury after inhibiting lentivirus expression of inflammation factor TNF-α. To study the relationship between Caspase-3, Bcl-2, Bax and TNF-α in spinal cord injury. Mthods Spinal cord contusion model was prepared by Allen method. The relation between tumor necrosis factor alpha and Bcl-2, was predicted by the method of GeneMANIA bioinformatics. The RNA which was packaged by lentivirus constructed the RNA interference model of tumour necrosis factor alpha. After interference of tumor necrosis factor alpha, we used the method of QRT-PCR to assays the mRNA expression of Caspase-3 and Bcl-2 in spinal cord and detect of the localization of Caspase-3 and Bcl-2 by immunohistochemistry. Statistical analysis with SPSS17.0. Results SD rats had paraplegia and urinate retentaion because of spinal cord injury. The result of QRT-PCR showed that in the seventh day after SCC, the expression of Caspase-3 reduced significantly (P < 0.05) and Bcl-2 did not change significantly (P > 0.05). Immunohistochemistry experiment results showed that Caspase-3 Bcl-2 and Bax immunoreactive cells were observed in the neurons and glial cells of both white matter and gray matter in the spinal cord. The results were the same with QRT-PCR.. Conclusion TNF-α in rats after SCC can effectively regulate the ratio of Bcl-2 and Bax , and then regulate the expression of Caspase-3 , which may affect the function of apoptosis and function recovery after spinal cord injury.