胺丁羟磷酸盐抑制皮肤角质形成细胞的增殖作用

Inhibitory effect of amphoteric alendronate on proliferation of skin keratinocytes

目的 探讨胺丁羟磷酸盐(ALD)对皮肤角质形成细胞(KCs)增殖的影响及中间产物是否拮抗其影响。方法 一定浓度的ALD、甲羟戊酸(MVA)、胆固醇(CH)、法尼基焦磷酸(FPP)、香叶基香叶基焦磷酸(GGPP)单独或联合处理KCs,细胞活力测定实验(MTS)法分析各处理组对KCs细胞增殖的影响;利用流式细胞术检测各加药组对细胞周期的影响;Western blot分析各加药组对细胞周期蛋白B1(Cyclin B1)、Cyclin E和细胞周期蛋白依赖性蛋白激酶抑制剂(P21)及磷酸化的蛋白激酶B(pAKT)和磷酸化的胞外信号调节激酶1/2(pERK1/2)表达量的影响。结果 MTS实验表明ALD对KCs增殖有显著抑制作用,而上游产物MVA与其有协同效应;下游产物中仅CH有部分补救作用。流式细胞术检测结果显示KCs被ALD阻滞在细胞分裂间期G1期,而仅CH可减弱ALD导致的G1期阻滞。Western blot结果表明ALD显著下调Cyclin B1,同时上调Cyclin E和P21的表达。此外,ALD能促进磷脂酰基酶3-激酶-蛋白激酶B(P13K-AKT)信号通路,抑制丝裂原激活蛋白激酶-胞外信号调节激酶(MAPKS-ERK1/2)信号通路的激活。结论 ALD显著抑制原代KCs的增殖,CH对这种抑制有部分的补救作用。

Objective To investigate the effects of alendronate(ALD)on the proliferation of skin keratinocytes(KCs)and to observe the effect of downstream products on this effect.Methods KCs were treated with a certain concentration of ALD,mevalonate(MVA),cholesterol(CH),farnesyl pyrophosphate(FPP),geranylgeranyl pyrophosphate(GGPP)alone or in combination MTS was performed to analyze the impact of ALD on the proliferation of KCs.Flow cytometry was used to detect the effect of each drug-added group on cell cycle.Western blot was used to analyze the level of Cyclin B1,Cyclin E and P21,pAKT and pERK1/2.Results MTS results showed that ALD significantly inhibited the proliferation of skin keratinocytes(KCs).Flow cytometry results of cell-cycle indicated that ALD caused G1 arrest.Only CH could rescue the proliferation inhibition and G1 arrest caused by ALD.Western blot results showed that ALD significantly down-regulated Cyclin B1 and up-regulated the expression of Cyclin E and P21.In addition,ALD could promote the P13 K-AKT signaling pathway and inhibit the activation of the MAPKS-ERK1/2 signaling pathway.Conclusion ALD significantly inhibits the proliferation of primary skin keratinocytes(KCs),and CH has a partial remedy for this inhibition.