CAJ | 학술논문

目的：研究远程缺血预处理（ RIPC）对大鼠脊髓缺血再灌注损伤（ SCII）后脑源性神经营养因子（ BDNF）表达的影响。方法2014年5—11月，将88只雄性SD大鼠按照随机数字表法分为假手术组（ Sham组， n＝8）、缺血再灌注组（I／R组， n ＝40）和RIPC组（n＝40）。 I／R组及RIPC组分别设再灌注0、6、12、24、72 h 5个观察时间点（分别记为1a组～1e组、2a组～2e组），每个时间点8只大鼠。 Sham组大鼠只分离肾动脉下腹主动脉，但不阻断； I／R组大鼠采用Zivin法建立SCII模型； RIPC组大鼠双下肢用驱血带短暂缺血10 min，放开10 min，重复2次，30 min后用Zivin法建立SCII模型。取材，苏木素-伊红（ HE）染色观察脊髓病理变化，免疫组化法检测BDNF阳性表达细胞数，原位细胞凋亡（ TUNEL）法检测细胞凋亡情况， Western blotting法检测BDNF表达水平，并进行大鼠后肢运动功能（ BBB）评分。结果 Sham组脊髓未见明显病理学改变， I／R组脊髓病理学改变明显， RIPC组各时间点脊髓病理学改变均较I／R组轻。1a组～1e组、2a组～2e组BDNF阳性表达细胞数、 BDNF表达水平均高于Sham组（P＜0.05）；1a组～1e组、2a组～2e组BBB评分均低于Sham组（P＜0.05）；1a组～1e组、2b组～2e组TUNEL阳性表达细胞数均高于Sham组（P＜0.05）；2b组～2e组BDNF阳性表达细胞数分别高于1b组～1e组（P＜0.05）；2a组～2e组BDNF表达水平、 BBB评分分别高于1a组～1e组（ P＜0.05）；2a组～2e组TUNEL阳性表达细胞数分别低于1a组～1e组（P＜0.05）；1b组～1e组BDNF阳性表达细胞数、 TUNEL阳性表达细胞数、 BDNF表达水平、 BBB评分均高于1a组（P＜0.05）；2b组～2e组BDNF阳性表达细胞数、 TUNEL阳性表达细胞数、 BDNF表达水平、 BBB评分均高于2a组（ P＜0.05）。结论 RIPC可增加大鼠SCII后BDNF的表达水平，并可能通过增加BDNF的表达水平来达到脊髓保护作用。目的：研究遠程缺血預處理（ RIPC）對大鼠脊髓缺血再灌註損傷（ SCII）後腦源性神經營養因子（ BDNF）錶達的影響。方法2014年5—11月，將88隻雄性SD大鼠按照隨機數字錶法分為假手術組（ Sham組， n＝8）、缺血再灌註組（I／R組， n ＝40）和RIPC組（n＝40）。 I／R組及RIPC組分彆設再灌註0、6、12、24、72 h 5箇觀察時間點（分彆記為1a組～1e組、2a組～2e組），每箇時間點8隻大鼠。 Sham組大鼠隻分離腎動脈下腹主動脈，但不阻斷； I／R組大鼠採用Zivin法建立SCII模型； RIPC組大鼠雙下肢用驅血帶短暫缺血10 min，放開10 min，重複2次，30 min後用Zivin法建立SCII模型。取材，囌木素-伊紅（ HE）染色觀察脊髓病理變化，免疫組化法檢測BDNF暘性錶達細胞數，原位細胞凋亡（ TUNEL）法檢測細胞凋亡情況， Western blotting法檢測BDNF錶達水平，併進行大鼠後肢運動功能（ BBB）評分。結果 Sham組脊髓未見明顯病理學改變， I／R組脊髓病理學改變明顯， RIPC組各時間點脊髓病理學改變均較I／R組輕。1a組～1e組、2a組～2e組BDNF暘性錶達細胞數、 BDNF錶達水平均高于Sham組（P＜0.05）；1a組～1e組、2a組～2e組BBB評分均低于Sham組（P＜0.05）；1a組～1e組、2b組～2e組TUNEL暘性錶達細胞數均高于Sham組（P＜0.05）；2b組～2e組BDNF暘性錶達細胞數分彆高于1b組～1e組（P＜0.05）；2a組～2e組BDNF錶達水平、 BBB評分分彆高于1a組～1e組（ P＜0.05）；2a組～2e組TUNEL暘性錶達細胞數分彆低于1a組～1e組（P＜0.05）；1b組～1e組BDNF暘性錶達細胞數、 TUNEL暘性錶達細胞數、 BDNF錶達水平、 BBB評分均高于1a組（P＜0.05）；2b組～2e組BDNF暘性錶達細胞數、 TUNEL暘性錶達細胞數、 BDNF錶達水平、 BBB評分均高于2a組（ P＜0.05）。結論 RIPC可增加大鼠SCII後BDNF的錶達水平，併可能通過增加BDNF的錶達水平來達到脊髓保護作用。
목적：연구원정결혈예처리（ RIPC）대대서척수결혈재관주손상（ SCII）후뇌원성신경영양인자（ BDNF）표체적영향。방법2014년5—11월，장88지웅성SD대서안조수궤수자표법분위가수술조（ Sham조， n＝8）、결혈재관주조（I／R조， n ＝40）화RIPC조（n＝40）。 I／R조급RIPC조분별설재관주0、6、12、24、72 h 5개관찰시간점（분별기위1a조～1e조、2a조～2e조），매개시간점8지대서。 Sham조대서지분리신동맥하복주동맥，단불조단； I／R조대서채용Zivin법건립SCII모형； RIPC조대서쌍하지용구혈대단잠결혈10 min，방개10 min，중복2차，30 min후용Zivin법건립SCII모형。취재，소목소-이홍（ HE）염색관찰척수병리변화，면역조화법검측BDNF양성표체세포수，원위세포조망（ TUNEL）법검측세포조망정황， Western blotting법검측BDNF표체수평，병진행대서후지운동공능（ BBB）평분。결과 Sham조척수미견명현병이학개변， I／R조척수병이학개변명현， RIPC조각시간점척수병이학개변균교I／R조경。1a조～1e조、2a조～2e조BDNF양성표체세포수、 BDNF표체수평균고우Sham조（P＜0.05）；1a조～1e조、2a조～2e조BBB평분균저우Sham조（P＜0.05）；1a조～1e조、2b조～2e조TUNEL양성표체세포수균고우Sham조（P＜0.05）；2b조～2e조BDNF양성표체세포수분별고우1b조～1e조（P＜0.05）；2a조～2e조BDNF표체수평、 BBB평분분별고우1a조～1e조（ P＜0.05）；2a조～2e조TUNEL양성표체세포수분별저우1a조～1e조（P＜0.05）；1b조～1e조BDNF양성표체세포수、 TUNEL양성표체세포수、 BDNF표체수평、 BBB평분균고우1a조（P＜0.05）；2b조～2e조BDNF양성표체세포수、 TUNEL양성표체세포수、 BDNF표체수평、 BBB평분균고우2a조（ P＜0.05）。결론 RIPC가증가대서SCII후BDNF적표체수평，병가능통과증가BDNF적표체수평래체도척수보호작용。
Objective To investigate the effect of remote ischemic preconditioning ( RIPC) on the expression of brain-derived neurotrophic factor (BDNF) in rats with spinal cord ischemia-reperfusion injury (SCII) .Methods From May to November in 2014, using random number table, 88 male Sprague Dawleys rats were randomly divided into three groups: the Sham group ( n=8 ) , the ischemia-reperfusion group ( I/R group, n=40 ) and the remote ischemic preconditioning group ( RIPC group, n=40) .For I/R group and RIPC group, 5 observation time points were set, including 0, 6, 12, 24 and 72 hours after reperfusion (designated as group 1a -1e for I/R group and group 2a -2e for RIPC group), 8 rats at each time point.For the rats in Sham group, only abdominal aorta below renal artery was isolated but without blocking; SCII model was built for the rats in I/R group by using Zivin method; for the rats in RIPC group, the lower limb blood flow was blocked with a tourniquet for 10 min and then was loosen for 10 min for reperfusion, which was done twice, and then SCII model was built using Zivin method 30 minutes later.Spinal cord was sampled from the rats.HE staining was used to observe pathological changes, the number of BDNF positive cells were detected by immunohistochemistry test, the cell apoptosis were assessed by TUNEL staining, Western blotting method was employed to detect the protein expression level of BDNF, and the BBB scores of the rats were evaluated.Results No evident pathological changes were noted in Sham group, while pathological changes were evident in I/R group, and the pathological changes in RIPC group were slighter than I/R group at five time points.Group 1a-1e and group 2a-2e were higher (P<0.05) than Sham group in the number of BDNF positive cells, the protein expression level of BDNF; group 1a-1e and group 2a-2e were lower (P<0.05) than Sham group in BBB score; group 1a-1e and group 2b-2e were higher (P<0.05) than Sham group in the number of TUNEL positive cells; group 2b-2e were higher (P<0.05) than group 1b-1e in the number of BDNF positive cells; group 2a-2e were higher (P<0.05) than group 1a-1e in the protein expression level of BDNF and BBB score; group 2a-2e were lower (P<0.05) than group 1a-1e in the number of TUNEL positive cells, group 1b-1e were higher (P<0.05) than group 1a in the number of BDNF positive cells, the number of TUNEL positive cells, the protein expression level of BDNF and BBB score; group 2b-2e were higher (P<0.05) than group 2a in the number of BDNF positive cells, the number of TUNEL positive cells, the protein expression level of BDNF and BBB score.Conclusion Remote ischemic preconditioning can increase the expression of BDNF in rats with spinal cord ischemia-reperfusion injury, and it may play a protective role through increasing the expression of BDNF.