摘要目的 通过观察牛磺酸对宫内生长受限( IUGR)胎鼠神经干细胞( NSCs)分化的影响,探讨其脑保护机制.方法 采用低蛋白饮食法建立IUGR胎鼠模型.取胎鼠室管膜下生发基质区( SVZ)脑组织进行NSCs培养.将传代培养所得NSCs分为3组:正常对照组、IUGR组和IUGR+牛磺酸组.分别对3组细胞进行诱导分化5 d.采用免疫荧光法检测各组分化细胞的神经细胞特异性微管蛋白β3( βⅢ-tubulin)、胶质纤维酸性蛋白(GFAP)、髓鞘碱性蛋白(MBP)阳性细胞数,计算各种细胞所占比例;采用Western blot法检测各组细胞βⅢ-tubulin及GFAP蛋白水平.结果 1.正常对照组、IUGR及IUGR+牛磺酸组βⅢ-tubulin阳性细胞计数比例分别为(18. 50 ± 0. 64)%、(15. 61 ± 0. 76)%、(18. 42 ± 0. 82)%,GFAP阳性细胞计数比例分别为(72. 19 ± 0. 82)%、(74. 87 ± 0. 67)%、(71. 68 ± 0. 92)%,组间比较差异均有统计学意义( F＝49. 103、44. 643,均 P<0. 01).与正常对照组比较,IUGR组βⅢ-tubulin阳性细胞计数比例显著减少( P<0. 01),GFAP阳性细胞计数比例显著增加(P<0. 01);与IUGR组相比,IUGR+牛磺酸组 βⅢ-tubulin阳性细胞计数比例显著增高( P<0. 01),GFAP阳性细胞计数比例显著减少( P<0. 01);正常对照组与IUGR+牛磺酸组二种细胞计数比例比较差异均无统计学意义(均P>0. 05).2.正常对照组、IUGR及IUGR+牛磺酸组βⅢ-tubulin蛋白相对表达量分别为0. 44 ± 0. 02、0. 33 ± 0. 03、0. 42 ± 0. 02,GFAP蛋白相对表达分别为1. 13 ± 0. 02、1. 50 ± 0. 04、1. 21 ± 0. 01,组间比较差异均有统计学意义( F＝45. 191、234. 525,均P<0. 01).与正常对照组相比,IUGR组βⅢ-tubulin蛋白表达显著减少(P<0. 01),GFAP蛋白水平显著增高(P<0. 01);与IUGR组相比,IUGR+牛磺酸组βⅢ-tubulin蛋白表达显著增高(P<0. 01),GFAP蛋白水平显著减少( P<0. 01);正常对照组与IUGR+牛磺酸组2种蛋白水平比较差异均无统计学意义(均P>0. 05).结论 牛磺酸可促进IUGR胎鼠NSCs向神经元方向分化,并保持各种分化细胞比例正常.

abstractsObjective To investigate the effect of Taurine on the differentiation of neural stem cells(NSCs)in fetal rats with intrauterine growth restriction( IUGR),and to explore the neuroprotective mechanism of Taurine. Methods IUGR fetal rats models were established with low protein diet. NSCs from subventricular zone( SVZ)were isolated and cultured in υitro. The NSCs were divided into 3 groups:normal control group,IUGR group,and IUGR+Taurine group. The cells were examined by adopting immunofluorescence for counting βⅢ-tubulin protein,glial fibril-lary acidic protein(GFAP)and myelin basic protein(MBP)-positive cells. Protein levels of βⅢ-tubulin and GFAP were detected by using Western blot. Results (1)The number of βⅢ-tubulin protein positive cells in normal control group,IUGR group and IUGR+Taurine group were(18. 50 ± 0. 64)%,(15. 61 ± 0. 76)% and(18. 42 ± 0. 82)%, respectively;the number of GFAP positive cells in normal control group,IUGR group and IUGR+Taurine group were (72. 19 ± 0. 82)%,(74. 87 ± 0. 67)% and(71. 68 ± 0. 92)%,respectively;and the differences were statistically sig-nificant(F＝49. 103,44. 643,all P<0. 01). Compared with the normal control group,βⅢ-tubulin protein positive cells in IUGR group decreased significantly(P<0. 01),but GFAP positive cells in IUGR group increased significantly (P<0. 01). Compared with IUGR Group,βⅢ-tubulin protein positive cells in IUGR+Taurine group increased sig-nificantly(P<0. 01),but GFAP positive cells in IUGR+Taurine group decreased significantly(P<0. 01). There was no significant difference between groupⅠand groupⅢ(all P>0. 05).(2)The levels of βⅢ-tubulin protein in nor- mal control group,IUGR group and IUGR+Taurine group were 0. 44 ± 0. 02,0. 33 ± 0. 03 and 0. 42 ± 0. 02,respective-ly;and the levels of GFAP protein in normal control group,IUGR group and IUGR+Taurine group were 1. 13 ± 0. 02, 1. 50 ± 0. 04,1. 21 ± 0. 01,respectively;and the differences were statistically significant(F＝45. 191,234. 525,all P<0. 01). Compared with normal control group,the levels of βⅢ-tubulin protein in IUGR group decreased significantly (P<0. 01),but the levels of GFAP in IUGR group increased significantly(P<0. 01). Compared with IUGR group, the levels of βⅢ-tubulin protein in IUGR+Taurine group increased significantly(P<0. 01),but the levels of GFAP in IUGR+Taurine group decreased significantly(P<0. 01). There was no significant difference between normal control group and IUGR+Taurine group(all P>0. 05). Conclusions Taurine can promote the differentiation of NSCs toward neurons in IUGR fetal rats,and maintain the normal proportion of all differentiated cells.