CAJ | 학술논문

背景研究表明糖尿病患者视网膜的神经病变出现在血管并发症之前,严重危害患者的视力,而神经营养因子对其具有保护作用.目的观察在糖尿病大鼠玻璃体腔内注射脑源性神经营养因子(BDNF)前后大鼠视网膜中BDNF及其受体TrkB、通路蛋白磷酸化细胞外信号调节激酶1/2( p-ERK1/2)、c-fos质量浓度的变化.方法 9周龄雄性健康Wistar大鼠按随机数字表法随机分为BDNF治疗组、糖尿病对照组和正常对照组,每组20只.BDNF治疗组、糖尿病对照组大鼠应用链脲佐菌素(STZ)腹腔注射制成糖尿病模型,BDNF治疗组大鼠于成模后2周开始向玻璃体腔内注射1 g/L BDNF溶液5μl.于成模后4周处死全部大鼠,取出眼球进行BDNF原位杂交,计数视网膜中BDNF阳性细胞数.采用 ELISA抗体夹心法检测视网膜中TrkB、p-ERK1/2以及c-fos蛋白质的质量浓度. 结果糖尿病对照组大鼠视网膜中BDNF阳性细胞数目明显少于正常对照组及BDNF治疗组,且染色浅,3个组BDNF阳性细胞数及阳性细胞灰度值比较差异均有统计学意义(F=102.36、92.55,P＜0.05).3个组TrkB、p-ERK1/2及c-fos表达的比较,差异均有统计学意义( F=92.54、95.46、94.84,P＜0.05).糖尿病对照组较正常对照组及BDNF治疗组TrkB质量浓度降低,而通路蛋白p-ERK1/2、c-fos质量浓度升高(P＜0.05)；正常对照组与BDNF治疗组间上述3个指标比较差异均无统计学意义(P＞0.05).结论在糖尿病早期,大鼠视网膜中BDNF mRNA水平及其受体TrkB蛋白质量浓度降低,其下游通路蛋白p-ERK1/2、c -fos质量浓度升高,玻璃体腔内注射BDNF可逆转上述变化.
배경 연구표명당뇨병환자시망막적신경병변출현재혈관병발증지전,엄중위해환자적시력,이신경영양인자대기구유보호작용.목적 관찰재당뇨병대서파리체강내주사뇌원성신경영양인자(BDNF)전후대서시망막중BDNF급기수체TrkB、통로단백린산화세포외신호조절격매1/2( p-ERK1/2)、c-fos질량농도적변화.방법 9주령웅성건강Wistar대서안수궤수자표법수궤분위BDNF치료조、당뇨병대조조화정상대조조,매조20지.BDNF치료조、당뇨병대조조대서응용련뇨좌균소(STZ)복강주사제성당뇨병모형,BDNF치료조대서우성모후2주개시향파리체강내주사1 g/L BDNF용액5μl.우성모후4주처사전부대서,취출안구진행BDNF원위잡교,계수시망막중BDNF양성세포수.채용 ELISA항체협심법검측시망막중TrkB、p-ERK1/2이급c-fos단백질적질량농도. 결과 당뇨병대조조대서시망막중BDNF양성세포수목명현소우정상대조조급BDNF치료조,차염색천,3개조BDNF양성세포수급양성세포회도치비교차이균유통계학의의(F=102.36、92.55,P＜0.05).3개조TrkB、p-ERK1/2급c-fos표체적비교,차이균유통계학의의( F=92.54、95.46、94.84,P＜0.05).당뇨병대조조교정상대조조급BDNF치료조TrkB질량농도강저,이통로단백p-ERK1/2、c-fos질량농도승고(P＜0.05)；정상대조조여BDNF치료조간상술3개지표비교차이균무통계학의의(P＞0.05).결론 재당뇨병조기,대서시망막중BDNF mRNA수평급기수체TrkB단백질량농도강저,기하유통로단백p-ERK1/2、c -fos질량농도승고,파리체강내주사BDNF가역전상술변화.
Background Recent studies showed that diabetic retinal neuropathy is an earlier and more dangerous complication and neurotrophin has a protective effect on retina. Objective The present study was to observe the changes of brain derived neurotrophic factor (BDNF),its receptor TrkB,signal pathway protein phosphatized extracellular signal-regulated protein kinase1/2 (p-ERK1/2) and c-fos in the retina after injection of BDNF into the vitreous in STZ induced Wistar diabetic rats. Methods Wistar rats aged 9 weeks-old were randomly divided into BDNF injection group,diabetes mellitus (DM) control group and normal control group and 20 rats for each group.STZ was intraperitoneally injected in the rats of BDNF injection group and DM control group to create the experimental DM.BDNF was intravitreously injected in the rats of BDNF group 2 weeks after administration of STZ in three-day interval for 5 times,and BSS containing O.1％ bovine serum albumin (BSA) was used at the same way in the DM control group and normal control group.The retina was isolated for hybridization in situ for BDNF,and TrkB,p-ERK1/2 and c-fos.Levels in retina were detected using sandwich method ELISA. Results The number of BDNF positive cells and the gray scale were lower obviously in the rat retina of DM control group than those of BDNF injection group and normal control group,showing significant differences among the 3 groups ( F =102.36,92.55 ;P＜0.05 ).ELISA assay showed that TrkB,p-ERK1/2 and c-fos values in retina were statistically significantly different among the 3 groups ( F =92.54,95.46,94.84,P＜0.05 ).The TrkB level in retina was statistically reduced,but the p-ERK1/2 and c-fos levels in retina were increased statistically in DM control group compared with BDNF injection group and normal control group( P＜0.05 ).No statistical difference was found in TrkB,p-ERK1/2 and c-fos values between the BDNF injection group and normal control group(P＞0.05). Conclusions The injection of BDNF into the vitreous cavity can protect retina from downregulating BDNF and TrkB levels and up-regulating the p-ERK1/2 and c-fos protein levels in the early stage of DM.