中华老年医学杂志
中華老年醫學雜誌
중화노년의학잡지
Chinese Journal of Geriatrics
2015年
8期
898-900
,共3页
李颖%毛瑞阳%杜晓红%赵晓薇
李穎%毛瑞暘%杜曉紅%趙曉薇
리영%모서양%두효홍%조효미
高同种半胱氨酸血症%白细胞介素-1β%肿瘤坏死因子α%NF-κB
高同種半胱氨痠血癥%白細胞介素-1β%腫瘤壞死因子α%NF-κB
고동충반광안산혈증%백세포개소-1β%종류배사인자α%NF-κB
Hyperhomocysteinemia%Interleukin-beta%Tumor necrosis factor-alpha%NF-kappaB
目的 探讨同型半胱氨酸诱导小胶质细胞表达白介素-1β(IL-1β)和肿瘤坏死因子α(TNF-α)的机制. 方法 将体外培养的小鼠小胶质瘤细胞(BV-2细胞)分成空白对照组、半胱氨酸(Cys)组、同型半胱氨酸(Hey)组和同型半胱氨酸+还原型谷胱甘肽(Hcy+GSH)组,培养72 h.应用反转录实时定量聚合酶链式反应(RT-qPCR)方法检测IL-1β和TNF-αmRNA表达,酶联免疫吸附法(ELISA)测定培养上清中细胞因子的蛋白浓度,蛋白质印迹法(Western blot)检测NF-κB/p65的含量. 结果 空白对照组、Cys组、Hcy组和Hcy+ GSH组细胞内IL-1β和TNF-α mRNA(F=48.63、130.76,均P＜0.05)和蛋白表达(F=702.91、293.69,均P＜0.05)以及NF-κB/p65含量(F=212.06,P＜0.05)差异均有统计学意义;与空白对照组比较,Hcy组细胞内IL-1β和TNF-α mRNA和蛋白表达明显增加,Hcy+GSH组较Hcy组表达水平下降,Hcy组NF-κB/p65含量高于其他组,差异均具有统计学意义(均P＜0.05). 结论 Hcy可诱导小胶质细胞表达IL-1β和TNF-α,NF-κB信号通路可能参与了此过程.
目的 探討同型半胱氨痠誘導小膠質細胞錶達白介素-1β(IL-1β)和腫瘤壞死因子α(TNF-α)的機製. 方法 將體外培養的小鼠小膠質瘤細胞(BV-2細胞)分成空白對照組、半胱氨痠(Cys)組、同型半胱氨痠(Hey)組和同型半胱氨痠+還原型穀胱甘肽(Hcy+GSH)組,培養72 h.應用反轉錄實時定量聚閤酶鏈式反應(RT-qPCR)方法檢測IL-1β和TNF-αmRNA錶達,酶聯免疫吸附法(ELISA)測定培養上清中細胞因子的蛋白濃度,蛋白質印跡法(Western blot)檢測NF-κB/p65的含量. 結果 空白對照組、Cys組、Hcy組和Hcy+ GSH組細胞內IL-1β和TNF-α mRNA(F=48.63、130.76,均P＜0.05)和蛋白錶達(F=702.91、293.69,均P＜0.05)以及NF-κB/p65含量(F=212.06,P＜0.05)差異均有統計學意義;與空白對照組比較,Hcy組細胞內IL-1β和TNF-α mRNA和蛋白錶達明顯增加,Hcy+GSH組較Hcy組錶達水平下降,Hcy組NF-κB/p65含量高于其他組,差異均具有統計學意義(均P＜0.05). 結論 Hcy可誘導小膠質細胞錶達IL-1β和TNF-α,NF-κB信號通路可能參與瞭此過程.
목적 탐토동형반광안산유도소효질세포표체백개소-1β(IL-1β)화종류배사인자α(TNF-α)적궤제. 방법 장체외배양적소서소효질류세포(BV-2세포)분성공백대조조、반광안산(Cys)조、동형반광안산(Hey)조화동형반광안산+환원형곡광감태(Hcy+GSH)조,배양72 h.응용반전록실시정량취합매련식반응(RT-qPCR)방법검측IL-1β화TNF-αmRNA표체,매련면역흡부법(ELISA)측정배양상청중세포인자적단백농도,단백질인적법(Western blot)검측NF-κB/p65적함량. 결과 공백대조조、Cys조、Hcy조화Hcy+ GSH조세포내IL-1β화TNF-α mRNA(F=48.63、130.76,균P＜0.05)화단백표체(F=702.91、293.69,균P＜0.05)이급NF-κB/p65함량(F=212.06,P＜0.05)차이균유통계학의의;여공백대조조비교,Hcy조세포내IL-1β화TNF-α mRNA화단백표체명현증가,Hcy+GSH조교Hcy조표체수평하강,Hcy조NF-κB/p65함량고우기타조,차이균구유통계학의의(균P＜0.05). 결론 Hcy가유도소효질세포표체IL-1β화TNF-α,NF-κB신호통로가능삼여료차과정.
Objective To investigate the mechanism of homocysteine-induced microglia (BV-2 cells) expression of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α).Methods The BV-2 cells were divided into blank control group,cysteine (Cys) group,homocysteine (Hcy) group and homocysteine and glutathione (Hcy+GSH) group,and the BV-2 cells in these groups were incubated with cysteine or homocysteine or homocysteine and glutathione together for 72 h.The mRNA expressions of IL-1β and TNF-α were assessed by RT-qPCR.The protein expressions of IL-1β and TNF-α in supernatant were detected by enzyme linked immunosorbent assay (ELISA).Western blot was used to observe the changes of NF-κB/p65 expression.Results There were significant differences in mRNA and protein expressions of IL-1β and TNF-α and NF-κB/p65 protein expression between groups (F=48.63,130.76,702.91,293.69,212.06,respectively,all P=0.000).The secretions of IL-1β and TNF-α were significantly improved by homocysteine (P＜0.05),and were reversed by the treatment with glutathione (P＜0.05).Western blot assay result showed that NF-κB/p65 was elevated after treatment with homocysteine (P＜0.05).Conclusions Homocysteine can induce microglia expression of interleukin-1β and tumor necrosis factor-α,and NF-κB signaling pathway may be involved in this process.
