EPCs在大鼠外伤性视神经损伤中变化的研究

发布时间：2018-04-27 00:01

  本文选题：视神经损伤 + 内皮祖细胞　； 参考：《天津医科大学》2011年硕士论文

【摘要】：目的： 1。建立Wistar大鼠外伤性视神经损伤动物模型,探讨不同损伤程度大鼠外伤性视神经损伤后外周血内皮祖细胞(endothelial progenitor cells,EPCs)的变化趋势及其与白细胞、血小板变化的关系。 2.分析外伤性视神经损伤对外周血内皮祖细胞的影响。 3.探讨内皮祖细胞在外伤性视神经损伤中的作用,为治疗外伤性视神经损伤提供理论依据。 方法： 1.采用液压冲击颅脑损伤仪建立外伤性视神经损伤动物模型,实验大鼠随机分为视神经损伤组(n=60)及正常对照组(n=20)；视神经损伤组依据打击力度不同分为轻度打击组(n=20),中度打击组(n=20),重度打击组(n=20)三个亚组。观察各组损伤前24h及损伤后3h、12h、24h、48h、72h、lw、2w、3w外周血EPCs、白细胞及血小板数量变化,同时观察HE染色变化。 2.采用液压冲击颅脑损伤仪制作外伤性视神经损伤动物模型,实验大鼠随机分为视神经损伤组和正常对照组；两组按照损伤前24h及损伤后3h、12h、24h、48h、72h、1w、2w、3w时间点随机分为9个亚组。测定各组上述时间点外周血EPCs数量并观察血管内皮标志物CD31免疫组化染色及F-VEP变化及微血管生成情况。 结果： 1.正常大鼠外周血EPCs数量为(46-52)个/20万单个核细胞,外伤性视神经损伤后外周血EPCs于伤后12h达最低点,各组间下降程度无显著差异(F=0.496, P0.05);轻度打击组及中度打击组EPCs数量在伤后24-72h升高至正正常水平以上,于伤后72h达峰,此后逐渐下降至正常水平；重度打击组EPCs于伤后24-72h虽有上升趋势,但达不到正正常水平；视神经损伤后大鼠外周血中EPCs数量变化与白细胞及血小板的数量无显著相关性(r=0.027,0.032,P0.05)。 2.正正常大鼠视神经及周围组织CD31+细胞数为(7-9)个／5个高倍视野,视神经损伤后各时间点创伤区CD31+细胞计数分别为(8.36±1.52、7.17±1.10、10.41±1.92、11.43±1.58.14.29±2.03、17.33±1.47、17.86±1.22、18.13±1.40)个／5个高倍视野,视神经损伤组与对照组比较,48h、72 h、1w、2w、3w时间点差异均有统计学意义(t=4.31,-7.61,-8.17,-10.08,-10.79;P0.05).正常大鼠视神经及周围组织微血管数量为(6～9)个/5个高倍视野,视神经损伤后各时间点损伤区微血管数量分别为(6.52±1.05.7.54±2.01.8.52±2.21. 11.02±1.62.15.40±2.04.18.39±1.96.23.21±1.50.22.78±2.40.24.13±2.51)个/5个高倍视野,视神经损伤组与对照组比较,48h.72 h.1w.2w.3w时间点差异均有统计学意义(t=4.25,-7.74,-8.26,-10.28,-11.49;P0.05).视神经损伤后各时间点F-VEP中P波潜伏期于损伤后3h降低,24h反弹增高到正常水平以上,并趋于稳定,视神经损伤组与对照组相比,3h.12h.24h. 48h.72h.1w.2w.3w差异均有统计学意义(t=4.15,3.74,5.84,6.08, 6.40,6.52,6.53,6.61；P0.05)；F-VEP中振幅于3h降低,12h升高到接近基础水平,24h后逐渐降低至基础水平以下,实验组与对照组比较,3h、24h.48h.72h.1w.2w.3w差异有统计学意义(t=3.95,4.14,5.26,5.78,6.49,6.72,6.23;P0.05)。外周血内皮祖细胞的数量变化与创伤区周围CD31+细胞、微血管、F-VEP潜伏期及振幅变化存在相关性(r=0.43,0.41,0.43,0.47；P0.05)。 结论： 1.外伤性视神经损伤大鼠外周血EPCs存在先降低后增高,再逐渐降至正常水平的变化特征。视神经损伤越严重,EPCs增高幅度越小。 2.外伤性视神经损伤能刺激外周血EPCs增加。 3.外伤性视神经损伤后增加的外周血EPCs定向归巢到创伤区,参与了创伤区血管新生和组织损伤修复。
[Abstract]:Purpose :

1 . To establish an animal model of traumatic optic nerve injury in Wistar rats . The changes of endothelial progenitor cells ( EPCs ) after traumatic optic nerve injury in rats with different degree of injury were investigated .

2 . To analyze the effect of traumatic optic nerve injury on endothelial progenitor cells in peripheral blood .

3 . To investigate the role of endothelial progenitor cells in traumatic optic nerve injury and to provide theoretical basis for the treatment of traumatic optic nerve injury .

Method :

1 . An animal model of traumatic optic nerve injury was established by hydraulic percussion brain injury instrument . The experimental rats were randomly divided into optic nerve injury group ( n = 60 ) and normal control group ( n = 20 ) .
The changes of EPCs , white blood cells and platelets in peripheral blood at 3h , 12h , 24h , 48h , 72h , lw , 2w , 3w were observed at 3h , 12h , 24h , 48h , 72h , lw , 2w , 3w before and after injury , and the changes of HE staining were observed .

2 . The experimental rats were randomly divided into optic nerve injury group and normal control group .
Two groups were randomly divided into 9 subgroups according to 24 h before injury and 3 h , 12 h , 24 h , 48 h , 72 h , 1 w , 2w , and 3w time points .

Results :

1 . The number of EPCs in peripheral blood of normal rats was ( 46 - 52 ) / 200,000 mononuclear cells . After traumatic optic nerve injury , the EPCs of peripheral blood reached the lowest point after injury ( F = 0.496 , P0.05 ) .
In severe attack group , the EPCs tended to increase in 24 - 72 h after injury , but reached normal level .
There was no significant correlation between the number of EPCs and the number of white blood cells and platelets in the peripheral blood of rats after optic nerve injury ( r = 0.027 , 0.032 , P0.05 ) .

2 . The number of CD31 + cells in optic nerve and peripheral tissues of normal rats was ( 7.17 卤 1.10 , 10.41 卤 1.92 , 11.43 卤 1.58.14 . 29 卤 2 . 03 , 17.33 卤 1 . 47 , 17.86 卤 1.22 , 18.13 卤 1.40 ) . The number of microvessels in the optic nerve and surrounding tissues of the normal rats was ( 6 - 9 ) / 5 high - fold visual field , and the number of microvessels at each time point after optic nerve injury was ( 6.52 卤 1.05 . 7.54 卤 2.01 . 8.52 卤 2 . 21.11 . 02 卤 1 . 96.23 . 21 卤 1 . 50.22 . 78 卤 1 . 96.23 . 21 卤 1 . 50 . 22 . 78 卤 2 . 40 . 24 . 13 卤 2 . 51 ) / 5 high - fold field of view , and the difference in the time point of optic nerve injury was significantly different from that of the control group ( t = 4.25 , - 7.74 , - 8.26 , - 10.28 , - 11.49 ; P0.05 ) . The latency of P wave in F - VEP after optic nerve injury was decreased at 3 h after injury , and the rebound increased to above normal level at 24 h , and tended to be stable . The difference of optic nerve injury group and control group was significantly ( t = 4.15 , 3.74 , 5.84 , 6.08 , 6.40 , 6.52 , 6.53 , 6.61 ) .
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