国际免疫学杂志
國際免疫學雜誌
국제면역학잡지
INTERNATIONAL JOURNAL OF IMMUNOLOGY
2010年
4期
320-324
,共5页
氨基甲酰促红细胞生成素%神经干细胞%氧糖剥夺
氨基甲酰促紅細胞生成素%神經榦細胞%氧糖剝奪
안기갑선촉홍세포생성소%신경간세포%양당박탈
Carbamylated erythropoietin%Neural stem cells%Oxygen glucose deprivation
目的 研究氨基甲酰促红细胞生成素(CEPO)对体外神经干细胞缺氧缺血性损伤的保护作用.方法 从孕14-16 d大鼠获得神经干细胞,经过培养并传代,对所获细胞的自我增殖、自我更新及多分化潜能进行检测.取第3代神经干细胞中添加CEPO,在氧糖剥夺(OGD)条件下培养2 h.通过TUNEL法计数神经干细胞凋亡率和MTT法检测神经干细胞的存活情况.采用酶联免疫吸附试验(ELISA)法检测神经干细胞分泌IFN-γ的情况.应用流式细胞仪检测MHC-Ⅱ在神经干细胞中的表达.结果 在OGD环境中,细胞凋亡率达58.97%,存活率为39.46%,IFN-γ和MHC-Ⅱ类分子的表达分别为78.47 pg/ml和35.68%,加入CEPO后神经干细胞的凋亡率下降至30.15%,存活率升高至75.84%,IFN-γ(15.35 pg/ml)和MHC-Ⅱ类分子的表达均下降(9.77%).结论 CEPO对神经干细胞缺氧缺血性损伤具有明显的保护作用,而且对炎症分子IFN-γ和免疫分子MHC-Ⅱ类分子的表达具有调节作用.
目的 研究氨基甲酰促紅細胞生成素(CEPO)對體外神經榦細胞缺氧缺血性損傷的保護作用.方法 從孕14-16 d大鼠穫得神經榦細胞,經過培養併傳代,對所穫細胞的自我增殖、自我更新及多分化潛能進行檢測.取第3代神經榦細胞中添加CEPO,在氧糖剝奪(OGD)條件下培養2 h.通過TUNEL法計數神經榦細胞凋亡率和MTT法檢測神經榦細胞的存活情況.採用酶聯免疫吸附試驗(ELISA)法檢測神經榦細胞分泌IFN-γ的情況.應用流式細胞儀檢測MHC-Ⅱ在神經榦細胞中的錶達.結果 在OGD環境中,細胞凋亡率達58.97%,存活率為39.46%,IFN-γ和MHC-Ⅱ類分子的錶達分彆為78.47 pg/ml和35.68%,加入CEPO後神經榦細胞的凋亡率下降至30.15%,存活率升高至75.84%,IFN-γ(15.35 pg/ml)和MHC-Ⅱ類分子的錶達均下降(9.77%).結論 CEPO對神經榦細胞缺氧缺血性損傷具有明顯的保護作用,而且對炎癥分子IFN-γ和免疫分子MHC-Ⅱ類分子的錶達具有調節作用.
목적 연구안기갑선촉홍세포생성소(CEPO)대체외신경간세포결양결혈성손상적보호작용.방법 종잉14-16 d대서획득신경간세포,경과배양병전대,대소획세포적자아증식、자아경신급다분화잠능진행검측.취제3대신경간세포중첨가CEPO,재양당박탈(OGD)조건하배양2 h.통과TUNEL법계수신경간세포조망솔화MTT법검측신경간세포적존활정황.채용매련면역흡부시험(ELISA)법검측신경간세포분비IFN-γ적정황.응용류식세포의검측MHC-Ⅱ재신경간세포중적표체.결과 재OGD배경중,세포조망솔체58.97%,존활솔위39.46%,IFN-γ화MHC-Ⅱ류분자적표체분별위78.47 pg/ml화35.68%,가입CEPO후신경간세포적조망솔하강지30.15%,존활솔승고지75.84%,IFN-γ(15.35 pg/ml)화MHC-Ⅱ류분자적표체균하강(9.77%).결론 CEPO대신경간세포결양결혈성손상구유명현적보호작용,이차대염증분자IFN-γ화면역분자MHC-Ⅱ류분자적표체구유조절작용.
Objective To observe the protection of carbamylated erythropoietin (CEPO) on neural stem cells (NSCs) impaired by oxygen glucose deprivation (OGD). Methods The NSCs of 14-16 days were isolated, cultured, and passaged. Proliferation, self-renewal ability and multipotency of differentiation of NSCs were examined. CEPO was added to the third passage of NSCs culture exposed to OGD condition for 2h. The apoptosis of NSCs was studied by TUNEL. The proliferation of NSCs was evaluated by MTT method. The level of IFN--γ was detected by ELJSA. Flow cytometry was applied to measure the expression of MHC-Ⅱ in NSCs. Results Under OGD condition, apoptosis of NSCs was 58.97% , and survival rate was 39.46% , and expressions of IFN--γ and MHC- Ⅱ were 78.47 pg/ml and 35.68% , respectively. Supplementation of CEPO displayed significant decrease of apoptosis (30. 15% ) and increase of survival rate (75. 84% ). The level of IFN-7 (15.35 pg/ml) and expression of MHC-H (9.77%) were decreased significantly in CEPO-treated group. Conclusion CEPO can significantly protect NSCs against hypoxia and ischemia, and meanwhile, CEPO can regulate the expression of IFN--γ and MHC- Ⅱ.
