摘要
目的探讨纳米脂质体槲皮素(nanoliposomal quercetin,nLQ)联合8-Br-cAMP对培养的人Rb44细胞组蛋白脱乙酰化酶的抑制效应。方法将培养的Rb44细胞分为4组:nLQ组,以40μmol·L-1nLQ培养;8-Br-cAMP组:以20μmol·L-18-Br-cAMP培养;nLQ+8-Br-cAMP组;以40μmol·L-1nLQ和20μmol·L-18-Br-cAMP共培养;对照组:不加nLQ或8-Br-cAMP。应用MTT实验检测细胞生长抑制率(GSR)。应用免疫斑点印迹检氉测脱乙酰化酶抑制物、NF-κBp65、磷酸化IκBα及c-myc的表达。对表达的靶信号以图像分析仪扫描灰度值。结果与对照组相比,nLQ组和8-Br-cAMP组GSR达44%±1%和40%±1%,均为P<0.05;nLQ+8-Br-cAMP组GSR达60%±1%,P<0.01。nLQ组、8-Br-cAMP组、nLQ+8-Br-cAMP组与对照组相比,脱乙酰化酶抑制物、NF-κBp65及c-myc的表达均减低,均为P<0.05;而磷酸化IκBα的表达均增强,均为P<0.05。结论 nLQ联合8-Br-cAMP可下调脱乙酰化酶抑制物表达,上调磷酸化IκBα的表达同时可抑制NF-κBp65的表达。
Objective To explore the inhibitory effects of nanoliposomal querce- tin（nLQ） combined with 8-Br-cAMP on histone deacetylase（HDAC） in human retino- blastoma Rb44 cells. Methods The cultured Rb44 cells were divided into 4 groups： nLQ group （cultured with 40 μmol · L^-1 nLQ ）, 8-Br-cAMP group （cultured with 20 μmol · L^-1 8-Br-cAMP） ,nLQ ＋ 8-Br-cAMP group（ cultured with 40μmol · L^-1 nLQ ＋ 20 μmol · L^-1 8-Br-cAMP） and control group （ cultured with neither nLQ nor 8-Br- cAMP）. The growth suppressing rate（GSR） was detected by MTY assay. The expression of HDAC, NF-KB p55, P （phosphorylated）-IKBu and c-myc were determined by immuno- cytochemistry and immunodotblotting, and the target signals were scanned with an im- age analyzer as grey scaled means. Results Compared with the control group, GSR in nLQ group and 8-Br-cAMP group were 44% ＋ 1% and 40% ＋ 1% （ both P 〈 0.05 ）, and nLQ ＋8-Br-cAMP group displayed synergistic effect, GSR was 50% ＋ 1% （P 〈 0.01 ）. Compared with control group,the expression of HDAC, NF-KB p55 and c-myc were de- creased in nLQ group, 8-Br-cAMP group and nLQ ＋ 8-Br-cAMP group （ P 〈 0.05 ）, while the expression of P-IKBα was enhanced（P 〈 0.05 ）. Conclusion nLQ combined with 8-Br-cAMP can inhibit HDAC expression acting as HDAC inhibitor, and up-regulate P- IκBα and inhibit NF-KB p55 expression.
出处
《眼科新进展》
CAS
北大核心
2012年第12期1110-1112,共3页
Recent Advances in Ophthalmology
基金
河南省科技攻关项目(编号:102300410047)~~
