摘要
背景:研究表明,Apelin/APJ在调节细胞增殖、凋亡、抑制炎症反应和血管重建等方面显示出功能。因此,推测Apelin在脊髓损伤中具有类似的功能。目的:评价Apelin对大鼠脊髓损伤的治疗作用。方法:①体内实验选用大鼠横断脊髓损伤模型,采用45只雌性SD大鼠,随机分为假手术组、脊髓损伤组、Apelin-13组。脊髓损伤组和Apelin-13组大鼠采用横断损伤法建立脊髓损伤模型,Apelin-13组每日腹腔注射Apelin-130.2 mg/kg,其余2组每日注射等量生理盐水,连续治疗14 d。于大鼠造模后第1,3,7,14天采用BBB运动评定量表评定大鼠后肢运动功能;于第14天收集大鼠脊髓,用于免疫组化、免疫荧光、RT-PCR等分析。②体外实验利用H2O2诱导PC12细胞损伤,加入不同浓度(1,2,4μmol/L)的Apelin-13,CCK8法检测其对诱导损伤的PC12细胞的活力的影响,探索Apelin的神经保护作用。结果与结论:①脊髓损伤后大鼠后肢功能完全丧失,损伤后3 d开始观察到运动恢复,但脊髓损伤组各时间点之间差异无显著性意义;Apelin促进了脊髓损伤大鼠的运动功能恢复;②脊髓损伤导致脊髓局部连续性破坏,Apelin促进了大鼠损伤脊髓形态的修复;③假手术组仅有少数离子钙接头蛋白分子1(iba1)阳性细胞,脊髓损伤导致局部该阳性细胞数量和星形胶质细胞显著增加,Apelin干预减少了局部星形胶质细胞和小胶质细胞激活(P<0.05);④脊髓损伤后严重脱髓鞘,LFB染色阳性面积明显降低,Apelin减少了脊髓损伤带来的脱髓鞘;⑤脊髓损伤后生长相关蛋白43表达量显著增加(P<0.001),给予Apelin进一步增加了生长相关蛋白43蛋白的表达;⑥Apelin增加了H2O2诱导的PC12损伤的细胞活力(P<0.05)。总之,Apelin促进了脊髓损伤大鼠运动和脊髓形态的修复。
BACKGROUND:Apelin/APJ exhibits functions in regulating cell proliferation,apoptosis,inhibiting inflammatory responses and vascular remodeling.Therefore,it is speculated that Apelin has similar functions in spinal cord injury.OBJECTIVE:To evaluate the effects of Apelin in rats with spinal cord injury.METHODS:A rat model of spinal cord transection was used in an in vivo experiment.Forty-five female Sprague-Dawley rats were randomly divided into sham group,spinal cord injury group and Apelin-13 group.Rats in the latter two groups were submitted to make animal models of spinal cord transection.Rats were given the intraperitoneal injection of 0.2 mg/kg Apelin-13 in the Apelin group or the same amount of normal saline in the other two groups for 14 consecutive days.The motor function of rat’s hind limbs was assessed by Basso,Beattie,Bresnahan locomotor rating scale at the 1^(st),3^(rd),7^(th),and 14^(th) days after modeling.The spinal cord tissues were collected at the 14th day after spinal cord injury and used for immunohistochemical,immunofluorescence and RT-PCR analyses.In in vitro experiments,H_(2)O_(2) was used to induce PC12 cell injury followed by treatment with different concentrations(1,2,4μmol/L)of Apelin-13.We then used cell counting kit-8 to detect the effect of Apelin-13 on the viability of injured PC12 cells,thereby exploring the neuroprotective effect of Apelin.RESULTS AND CONCLUSION:(1)After spinal cord injury,the hindlimb function of the rats was completely lost and motor recovery was observed 3 days after injury.However,there was no significant difference in the spinal cord injury group at different time points.Apelin promoted motor function recovery post spinal cord injury.(2)Spinal cord injury disrupted the local continuity of the spinal cord,and Apelin restored the injured morphology induced by spinal cord injury.(3)There were only a few cells positive for ionized calcium binding adapter protein 1 in the sham group.Spinal cord injury led to a sharp increase in the number of local positive cells and astrocytes.Apelin could alleviate the local activation of astrocytes and microglia(P<0.05).(4)Severe demyelination occurred after spinal cord injury and the positive area of Luxol Fast Blue staining was significantly reduced.Apelin could attenuate demyelination caused by spinal cord injury.(5)The expression of growth-related protein 43 was significantly increased after spinal cord injury(P<0.001).Apelin could further promote the expression of growth-related protein 43(P<0.05).(6)Apelin increased the cell viability of H2O2-stimulated PC12 cells in vitro(P<0.05).To conclude,Apelin promotes locomotor recovery and spinal cord morphology repair in rats after spinal cord injury.
作者
刘卿
王笑
谷成旭
郭绮萱
李希凯
张璐萍
黄飞
Liu Qing;Wang Xiao;Gu Chengxu;Guo Qixuan;Li Xikai;Zhang Luping;Huang Fei(Department of Anatomy,Basic Medical University,Yantai 256699,Shandong Province,China;School of Health and Life Sciences,University of Health and Rehabilitation Science,Qingdao 266071,Shandong Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2023年第11期1744-1749,共6页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金(81870985,81171142),项目负责人:黄飞。
