背景研究表明，实验性外伤性视神经病变(TON)可激活机体的免疫系统而发生免疫性炎症反应，而一种CD4 ＋ T淋巴细胞，即辅助性T细胞17(Th17)与免疫性炎症的发生和发展有关，但Th17与TON发生和发展的关系尚不明确。
目的观察TON大鼠脾脏中Th17细胞数量及视网膜中白细胞介素-17(IL-17)的表达变化。
方法采用随机数字表法将70只4周龄雄性健康SD大鼠分为正常对照组、伪手术组和TON 1、3、7、14、28 d组，每组10只。TON组大鼠右眼结膜切开后分离并暴露视神经，用液压颅脑损伤(FPI)仪以25°击打视神经，平均打击力为(699.1±60.8)kPa，建立TON大鼠模型，伪手术组大鼠仅暴露视神经。各组大鼠均行闪光视觉诱发电位(F-VEP)检查，测量P 2波的潜伏期和振幅；收集各组大鼠脾脏以制备单细胞悬液，采用流式细胞技术计数Th17细胞；采用免疫组织化学法检测各组大鼠视网膜组织中IL-17蛋白的表达。
结果正常对照组与伪手术组间大鼠F-VEP P 2波潜伏期和振幅、脾脏中Th17细胞占CD4 ＋ T细胞的比例、视网膜组织中IL-17阳性细胞数比较，差异均无统计学意义( P＝0.829、0.830、0.856、0.496)。与伪手术组比较，各TON组大鼠F-VEP P 2波的潜伏期明显延长，振幅明显下降，差异均有统计学意义(均 P<0.05)。TON 1、3、7、14和28 d组大鼠脾脏中Th17细胞占CD4 ＋ T细胞的比例分别为(1.80±0.18)%、(1.98±0.20)%、(2.34±0.20)%、(2.11±0.13)%、(1.92±0.18)%，均明显高于伪手术组的(0.94±0.13)%，差异均有统计学意义(均 P<0.05)。TON 1、3、7、14和28 d组大鼠视网膜组织中IL-17阳性细胞数分别为1 673.416±188.021、1 892.431±151.026、2 420.454±256.024、1 996.429±177.022和1 629.410±127.023，均明显高于伪手术组的796.326±100.028，差异均有统计学意义(均 P<0.05)。
结论TON大鼠脾脏中Th17细胞占CD4 ＋ T细胞的比例上升，视网膜组织中IL-17蛋白表达量增加，二者可能参与TON的发生和发展过程。
BackgroundStudies showed that traumatic optic neuropathy (TON) can activate and induce immuno-inflammatory response, while T helper cell 17 (Th17), a CD4 + T lymphocyte, is associated with immuno-inflammatory response.However, the effects of Th17 on the development of TON are unclear.
ObjectiveThis study was to observe the changes of Th17 lymphocytes in spleen and the expression of interleukin-17 (IL-7) in retina in TON rats.
MethodsSeventy male 4-week-old SD rats were randomly divided into normal control group, sham operation group and TON 1-, 3-, 7-, 14- and 28-day group according to random number table.The optical nerves of the right eyes were exposed and impacted at a 25° angle by using fluid percussion injury (FPI) device with the average force (699.1±60.8) kPa to create the TON models in the TON groups, and only optical nerves were exposed in the sham operation group.Flash visual evoked potential (F-VEP) was recorded and the amplitude and latency were measured to evaluate the function of optical nerves of the rats.The rat splenocyte suspension in various groups was prepared for the detection of Th17 cells by flow cytometry, and the expression of IL-17 protein was detected by immunochemistry.
ResultsNo significant differences were found in the amplitudes and latencies of P 2 wave, proportion of Th17 cells and the IL-17 positive cell numbers in retina between the normal control group and sham operation group ( P=0.829, 0.830, 0.856, 0.496). The amplitudes of P 2 wave were significantly lower, and the latencies of P 2 wave were significantly longer in the TON groups than those in the sham operation group (all at P<0.05). The proportions of Th17 positive CD4 + T lymphocytes were (0.94±0.13)%, (1.80±0.18)%, (1.98±0.20)%, (2.34±0.20)%, (2.11±0.13)%, (1.92±0.18)% in the sham operation group, TON 1-day group, TON 3-day group, TON 7-day group, TON 14-day group and TON 28-day group, respectively, showing significant differences between the sham operation group and various TON groups (all at P<0.05). In addition, the IL-17 positive cell numbers in retina were 796.326±100.028, 1 673.416±188.021, 1 892.431±151.026, 2 420.454±256.024, 1 996.429±177.022 and 1 629.410±127.023 in the sham operation group, TON 1-day group, TON 3-day group, TON 7-day group, TON 14-day group and TON 28-day group, respectively, showing significant differences between the sham operation group and various TON groups (all at P<0.05).
ConclusionsIn rats of TON, the proportion of Th17 cell in splenocytes and the expressions of IL-17 in retina are increased, suggesting that Th17 and IL-17 participate in the progression of TON.
郑华宾,颜华. 实验性外伤性视神经病变大鼠脾细胞中Th17细胞数量与视网膜中IL-17表达的变化[J]. 中华实验眼科杂志,2015,33(10)：876-880.
DOI:10.3760/cma.j.issn.2095-0160.2015.10.003版权归中华医学会所有。
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