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目的探讨甲状腺激素诱导的神经干细胞(NSCs)是否比单纯NSCs移植对慢性实验性变态反应性脑脊髓炎(EAE)有更好的神经保护作用。方法体外培养新生大鼠NSCs和三碘甲腺原氨酸(T3)诱导的NSCs(T3/NSCs),诱导其分化7d后,免疫细胞化学或免疫荧光染色分别检测半乳糖脑苷脂阳性(Gal C+)和GFAP阳性(GFAP+)细胞。用豚鼠脊髓匀浆诱导慢性EAE大鼠模型。在免疫后10d,脑立体定位仪分别移植T3/NSCs、NSCs和生理盐水入EAE大鼠侧脑室,T3/NSCs和NSCs移植前用5-溴脱氧尿嘧啶核苷(Brd U)标记。实验分为T3/NSCs组、NSCs组和对照组,每组10只。每天对大鼠临床症状评分评估神经功能。大鼠免疫60d后处死,HE和LFB染色分别观察脑的炎症侵润和脱髓鞘;免疫荧光双标染色检测脑的Gal C+/Brd U+和GFAP+/Brd U+比例;RT-PCR法检测脑组织血小板源性生长因子α受体(PDGFαR)、Gal C和髓鞘碱性蛋白(MBP)mRNA的表达。结果 T3/NSCs体外分化为Gal C+和GFAP+细胞的比例分别高于和低于NSCs的分化。T3/NSCs组和NSCs组神经功能恢复较对照组好。T3/NSCs组较NSCs组更明显改善脑的炎症侵润和脱髓鞘,其脑内Gal C+/Brd U+及GFAP+/Brd U+的比例分别高于和低于NSCs组,脑组织的PDGFαR、Gal C和MBP mRNA的表达也较NSCs组高。结论 T3/NSCs比NSCs对EAE有更好的神经保护作用。
Objective To investigate whether thyroid hormone-induced neural stem cells (NSCs) have a better neuroprotective effect than chronic NSA transplantation on chronic experimental allergic encephalomyelitis (EAE). Methods NSCs (T3 / NSCs) induced by neonatal rat NSCs and triiodothyronine (T3) were cultured in vitro and were induced to differentiate for 7 days. Immunocytochemistry or immunofluorescence staining were used to detect the expression of galactocerebrosidase C +) and GFAP-positive (GFAP +) cells. Chronic EAE rat model was induced by guinea pig spinal cord homogenate. Ten days after the immunization, brain stereotaxic apparatus was transplanted with T3 / NSCs, NSCs and normal saline into lateral ventricle of rats, and T3 / NSCs and NSCs were labeled with BrdU before transplantation. The experiment was divided into T3 / NSCs group, NSCs group and control group, 10 in each group. The clinical symptoms of rats were evaluated daily for neurological function. The rats were sacrificed 60 days after immunization and the inflammatory infiltration and demyelination were observed by HE and LFB staining respectively. The ratios of Gal C + / Brd U + and GFAP + / Brd U + were measured by double immunofluorescence staining. The brain tissues Expression of platelet-derived growth factor alpha receptor (PDGFαR), Gal C and myelin basic protein (MBP) mRNA. Results The ratio of T3 / NSCs differentiated into Gal C + and GFAP + cells in vitro was higher than that of NSCs. The neurological function recovery of T3 / NSCs group and NSCs group was better than that of control group. The ratio of Gal C + / Brd U + and GFAP + / Brd U + in the brain of the T3 / NSCs group was significantly higher than that of the NSCs group in the inflammatory infiltration and demyelination. The ratio of PDGFαR, Gal The expression of C and MBP mRNA was also higher than that of NSCs. Conclusion T3 / NSCs have better neuroprotective effects on EAE than NSCs.