人参皂甙Rg1促进嗅鞘细胞修复脊髓损伤的研究

Ginsenosides Rg1 enhance the ability of olfactory ensheathing cells to repair spinal cord injury： An experimental study

目的 观察人参皂甙Rg1对嗅鞘细胞（OECs）移植修复脊髓损伤能力的影响.方法 新生SD大鼠嗅球组织体外培养、纯化OECs,人参皂甙Rg1干预并设置对照,Allen打击建立大鼠脊髓损伤模型,随机分成3组：对照组（Con组）、OECs移植组（OECs组）和Rg1干预OECs移植组（Rg1＋OECs组）,于移植后第1、3、7、14、28天分别进行肢体活动大鼠脊髓损伤[Basso-Beattie-Bresnahan（BBB）]评分,并于第28天取损伤部位脊髓组织制作石蜡切片进行免疫组织化学检测;反转录-聚合酶链反应（RT-PCR）分别测定OECs组和Rg1＋ OECs组脊髓组织中睫状神经营养因子（CNTF）和血管内皮生长因子（VEGF）的mRNA表达.结果 Rg1＋ OECs组大鼠后肢运动恢复情况优于OECs组和对照组[（15.0 ±1.0）、（12.0±2.0）、（7.4±0.9）分,P＜0.05],组织切片示Rg1＋ OECs组神经元数量明显高于OECs组和对照组[（64±3）、（38±2）、（26±2）个,P＜0.01],胶质纤维酸性蛋白（GFAP）表达量（以GFAP染色阳性细胞平均积分吸光度值作为检测指标）显著低于OECs组和对照组（8.65±0.32、4.93±0.25、2.21±0.17,P＜0.01）,苏木素-伊红（HE）染色示Rg1＋ OECs组脊髓组织结构清晰,囊性病变及细胞水肿坏死数量明显少于其他各组;RT-PCR结果显示人参皂甙Rg1显著上调OECs在移植宿主体内CNTF、VEGF mRNA表达（0.78±0.03、0.39±0.03;0.86±0.03、0.61±0.02,P＜0.05）.结论 人参皂甙Rg1能够通过上调OECs在宿主体内CNTF、VEGF的表达,提高OECs移植修复脊髓损伤的能力.

Objective To study the effects of ginsenoside Rg1 on effects of olfactory ensheathing cells （OECs） in spinal cord injury as well as their expression of neurotrophic factors like ciliary neurotrophic factor （CNTF）,vascular endothelial growth factor （VEGF）.Methods OECs were primary cultured and purified from olfactory bulb of newborn SD rats.Purified cells were divided into 2 groups,the experimental group was treated with Rg1 in concentration of 40 μg/ml for 72 h （the optimal concentration and action time which have been proven by our previous study）,the control group received an equal volume of Dulbecco＆#39; s Modified Eagle Media：Nutrient Mixture F-12 （DMEM/F12） medium.The models of spinal cord injury was established by using Allen＆#39; s combat,and were randomly divided into 3 groups,OECs group （inject the control group cells）,Rg1 ＋ OECs group （inject the experimental group cells） and control group （inject an equal volume of DMEM/F12 medium）.Using reverse transcriptase-polymerase chain reaction （RT-PCR） to determine mRNA expression of CNTF and VEGF in OECs group and Rg1 ＋ OECs group.Results The Basso Beattie Bresnahan （BBB） score of Rg1 ＋ OECs group was higher than the other two groups （15.0 ± 1.0 vs.12.0 ± 2.0,7.4 ± O.9,P 〈 0.05）.Histology revealed that Rg1 ＋ OECs group had larger number of neurons （64 ± 3,38 ± 2,26 ± 2,P 〈 0.01）,lower level of GFAP expression and less damage of spinal cord tissue than OECs group and control group （8.65 ± 0.32,4.93 ± 0.25,2.21 ± 0.17,P 〈 0.01）.RT-PCR analysis demonstrated that Rg1 significantly up-regulated mRNA expression of CNTF and VEGF （0.78 ±0.03,0.39 ±0.03;0.86 ±0.03,0.61±0.02,P 〈0.05）.Conclusion Ginsinoside Rg1 obviously improve the effects of olfactory ensheathing cell transplantation for spinal cord injury by the up-regulation of CNTF and VEGF.