摘要
背景:研究表明紫杉醇可以治疗骨转移肿瘤,说明紫杉醇对于治疗骨破坏疾病有一定的疗效,但是目前国内外对于紫杉醇治疗骨破坏相关疾病的研究很少,关于紫杉醇调节破骨细胞形成和功能的机制仍然不明确。目的:观察紫杉醇对根尖周炎破骨细胞增殖及分化的影响。方法:体外培养RAW264.7细胞24 h后换RANKL诱导液培养,通过Real-time PCR及抗酒石酸酸性磷酸酶鉴定破骨细胞诱导成功。CCK8检测不同浓度(1,10^(-1),10^(-2),10^(-3),10^(-4),10^(-5),10^(-6) mol/L)的紫杉醇作用后破骨细胞增殖情况,筛选紫杉醇作用于破骨细胞最佳浓度;以未加入紫杉醇的破骨细胞为对照组;采用Real time-PCR和Western blot检测紫杉醇对破骨细胞凋亡的影响。结果与结论:(1)紫杉醇10^(-4) mol/L时,破骨细胞增殖能力开始降低(P <0.05),表明10^(-4) mol/L紫杉醇为作用于RAW264.7细胞的最佳浓度;(2)加入10^(-4) mol/L紫杉醇0,24和48 h,破骨细胞增殖均显著低于未加入紫杉醇对照组(P<0.05);破骨细胞抗酒石酸酸性磷酸酶A值的增殖均显著低于未加入紫杉醇对照组(P<0.05);(3)Realtime-PCR检测结果显示,加入10^(-4)mol/L紫杉醇抗酒石酸酸性磷酸酶m RNA表达明显低于未加紫杉醇的对照组(P <0.05),Caspase3和PARP1的蛋白表达明显高于未加紫杉醇的对照组;(4)结果说明,紫杉醇抑制破骨细胞的增殖及分化,并且可以促进破骨细胞的凋亡。
BACKGROUND:Paclitaxel has been shown to treat bone metastases,suggesting that it has certain effect on osteolysis diseases.However,there are few studies on paclitaxel for osteolysis diseases,and the mechanism of paclitaxel regulating osteoclast formation and function remains unclear.OBJECTIVE:To investigate the effect of paclitaxel on the proliferation and differentiation of osteoclasts in periapical periodontitis.METHODS:RAW264.7 cells were cultured in vitro for 24 hours and RANKL was added to identify osteoclasts by real-time PCR and tartrate-resistant acid phosphatase.Different concentrations(1,10-1,10-2,10-3,10-4,10-5,10-6 mol/L)of paclitaxel were used to treat osteoclasts,and the optimal concentration of osteoclasts was selected by cell counting kit-8 assay.The osteoclasts without paclitaxel were used as controls.The effect of paclitaxel on osteoclast apoptosis was detected by real-time PCR and western blot assay.RESULTS AND CONCLUSION:The osteoclasts induced by 10-4 mol/L paclitaxel showed the weakest proliferation ability(P<0.05),suggesting that 10-4 mol/L was the optimal concentration.After cultured by 10-4 mol/L paclitaxel for 0,24 and 48 hours,the proliferation of osteoclasts was significantly lower than that in the control group(P<0.05).The absorbance value of tartrate-resistant acid phosphatase of the osteoclasts cultured by 10-4 mol/L paclitaxel was significantly lower than that in the control group(P<0.05).Real-time PCR results revealed that mRNA expression level of tartrate-resistant acid phosphatase of the osteoclasts cultured by 10-4 mol/L paclitaxel was significantly lower than that in the control group(P<0.05).The protein expression levels of Caspase3 and PARP1 of the osteoclasts cultured by 10-4 mol/L paclitaxel were significantly higher than those in the control group.In summary,paclitaxel can inhibit the proliferation and differentiation of osteoclasts,and activate apoptosis of osteoclasts.
作者
吴慧
姜龙
李晓杰
季秋实
许诺
Wu Hui;Jiang Long;Li Xiaojie;Ji Qiushi;Xu Nuo(Department of Stomatology,the First Affiliated Hospital of Hainan Medical University,Haikou 570102,Hainan Province,China;Zhongshan College of Dalian Medical University,Dalian 116000,Liaoning Province,China;Stomatology College of Dalian Medical University,Dalian 116044,Liaoning Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2019年第11期1647-1651,共5页
Chinese Journal of Tissue Engineering Research
基金
辽宁省科学技术计划项目面上项目(201600752)
项目负责人:李晓杰~~
