《中国康复理论与实践》

《中国康复理论与实践》 ›› 2012, Vol. 18 ›› Issue (6): 530-534.

• 论文 • 上一篇    下一篇

不同剂量FTY720 对大鼠急性脊髓损伤后Caspase-3 表达及细胞凋亡的影响

杨梁1，吕德成2，郑连杰1，王朝晖1，张伟3，李晓天3   

  1. 1.大连医科大学附属二院骨科，辽宁大连市116027；2.大连医科大学附属一院骨科，辽宁大连市116011；3.大连医科大学，辽宁大连市116021。
  • 收稿日期:2012-02-14 修回日期:1900-01-01 出版日期:2012-06-25 发布日期:2012-06-25

Effects of Different Doses of FTY720 on Expression of Caspase-3 and Neural Apoptosis in Rats with Acute Spinal Cord Injury

YANG Liang, LÜ De-cheng, ZHENG Lian-jie, et al.   

  1. Department of Orthopaedics, Second Affiliated Hospital, Dalian Medical University, Dalian 116027, Liaoning, China
  • Received:2012-02-14 Revised:1900-01-01 Published:2012-06-25 Online:2012-06-25

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被引次数

9

摘要： 目的比较不同剂量FTY720 对大鼠急性脊髓损伤后Caspase-3 的表达及神经细胞凋亡的影响。方法雌性SD 大鼠200 只，随机分为5 组，每组40 只。A 组大鼠不打击脊髓，缝合切口后立即以0.3 ml 生理盐水灌胃。B、C、D、E 组大鼠制作Allen'sT9脊髓损伤模型。B组造模后立即以0.3 ml 生理盐水灌胃，C、D、E 组分别以FTY720 按1 mg/kg、3 mg/kg、5 mg/kg 生理盐水稀释至0.3 ml 灌胃。分别于术后6 h、12 h、24 h、48 h 和72 h 取损伤段脊髓超薄切片，行SP 免疫组化染色观察Caspase-3 表达及TUNEL 染色观察细胞凋亡情况，并计算相应时间点免疫组化学染色阳性细胞比值。结果A组各时间点几乎见不到Caspase-3 和细胞凋亡阳性表达。B、C、D、E 组均在伤后6 h 开始出现凋亡细胞表达，伤后24 h 达高峰，伤后48 h 开始减弱，伤后72 h 进一步减少。伤后各时间点细胞凋亡表达均为B 组>C 组>D 组=E 组(P>0.05)>A 组(P<0.05)。Caspase-3 表达与细胞凋亡同步。相关性检验显示B、C、D组中，FTY720 的剂量与Caspase-3 表达、细胞凋亡表达呈负相关(P<0.05)。D组和E 组中，FTY720 剂量与上述指标不相关(P>0.05)。结论FTY720 可以显著减少大鼠急性脊髓损伤后Caspase-3 的表达及神经细胞凋亡。FTY720 的剂量与其减轻Caspase-3 表达和细胞凋亡效果之间存在一定量效关系。3 mg/kg 是FTY720 治疗大鼠急性脊髓损伤的最适剂量。

关键词: FTY720, Caspase-3, 凋亡, 脊髓损伤, 神经再生, 剂量, 量效关系

Abstract: Objective To compare the effects of different doses of FTY720 on inhibiting expression of Caspase-3 and neural apoptosisin rats with acute spinal cord injury (SCI), and find out the suitable dose. Methods 200 female SD rats were randomly divided into 5 groupswith 40 in each group. Group A (laminectomy but not contusion) were administered 0.3 ml normal saline by gavage. SCI model was establishedby Allen's WD method at the T9 level of spinal cord in other groups. Group B were administered 0.3 ml normal saline after modeling.Groups C, D and E were administered 0.3 ml FTY720- saline solution of 1, 3 and 5 mg/kg respectively. All the groups were sacrificed at 6 h,12 h, 24 h, 48 h, 72 h (n=8, per each time-point). Caspase-3 expression was detected with streptavidin-peroxidase immunohistochemistry,and neural apoptosis was detected with the TUNEL method. Results Positive Caspase-3 expression and neural apoptosis were not observedin Group A at 6 h. In Groups B、C、D and E, the number of apoptotic cells increased with increased time of acute SCI, peaked at 24 h afterinjury, and then gradually reduced. Caspase-3 expression was at equal pace with neural apoptosis. The difference of the number of apoptoticand Caspase-3 expression cells among all groups were significant, with the order Group B>Group C>Group D>Group A (P<0.05). However,there was no significant difference between Group D and Group E (P>0.05). The number of apoptotic and Caspase-3 expression cells negativelycorrelated with the dose of FTY720 when the dose was less than 3 mg/kg (P<0.05), and there was no relationship when the dose wasmore than 3 mg/kg (P>0.05). Conclusion FTY720 significantly reduces Caspase-3 expression and neural apoptosis in rats with acute SCI.There is a dose-effect relationship between the dose of FTY720 and the Caspase-3 expression and neural apoptosis. It's indicated that 3 mg/kg is the most appropriate dosage.

Key words: FTY720, Caspase-3, apoptosis, spinal cord injury, neural regeneration, dose, dose-effect relationship