多巴胺毒性代谢产物对前成骨细胞的增殖、分化、矿化及凋亡的影响  

Effects of Dopamine Toxic Metabolites on the Proliferation, Differentiation, Mineralization and Apoptosis of Pre-osteoblasts

作　　者：李颖 吴玉波[1] Li Ying;Wu Yubo(Department of Clinical Pharmacy,Fourth Affiliated Hospital of Harbin Medical University,Harbin 150001,China)

机构地区：[1]哈尔滨医科大学附属第四医院临床药学科,哈尔滨150001

出　　处：《中国药师》2021年第12期2182-2187,共6页China Pharmacist

摘　　要：目的:探究多巴胺的毒性代谢产物3,4-二羟基苯乙醛(DOPAL)对前成骨细胞增殖、分化、矿化及凋亡的影响及其可能的作用机制。方法:培养MC3T3-E1前成骨细胞,将细胞分为对照组及不同浓度的DOPAL加药组,采用CCK-8检测DOPAL对前成骨细胞增殖的影响;qRT-PCR检测成骨细胞标志因子mRNA相对表达;茜素红染色检测其对成骨细胞骨矿化结节的影响;Tunel及Western Blot检测DOPAL对前成骨细胞凋亡的影响;Western Blot检测α-突触核蛋白(α-S)蛋白相对表达。结果:当DOPAL的加药浓度≥0.1μmol·L^(-1)时,对MC3T3-E1前成骨细胞增殖有显著的抑制作用,与对照组比较,差异有统计学意义(P<0.05或P<0.01)。与对照组相比,DOPAL各浓度组的成骨细胞标志因子mRNA相对表达显著下调(P<0.05或P<0.01),细胞凋亡标志因子cleaved-caspase-3蛋白相对表达、细胞凋亡率、α-S蛋白相对表达显著升高(P<0.01);DOPAL 0.1μmol·L^(-1)和0.5μmol·L^(-1)两个浓度组以上各指标差异有统计学意义(P<0.05或P<0.01)。未加入骨诱导剂的对照组,并未出现矿化结节;与骨诱导组相比,DOPAL 0.1μmol·L^(-1)和0.5μmol·L^(-1)组抑制MC3T3-E1细胞分化成熟。结论:DOPAL抑制前成骨细胞的增殖、分化及矿化作用,且促进前成骨细胞的凋亡,可能与前成骨细胞中α-S蛋白相对表达增加从而聚集形成有神经毒性的寡聚体有关,该实验为在体研究奠定基础。Objective: To explore the effects of 3,4-dihydroxyphenylacetic acid(DOPAL), a toxic metabolite of dopamine, on the proliferation, differentiation, mineralization and apoptosis of pre-osteoblasts and its possible mechanism. Methods: MC3 T3-E1 pre-osteoblasts were cultured, and the cells were divided into control group and DOPAL plus drug group with different concentrations(0.03,0.1,0.3,1 μmol·L^(-1)). CCK-8 was used to detect the effect of DOPAL on the proliferation of pre-osteoblasts;qRT-PCR was used to detect the relative expression of osteoblast marker factor mRNA;alizarin red staining was used to detect its effect on bone mineralization nodules of osteoblasts;Tunel and Western blot were used to detect the effect of DOPAL on the apoptosis of pre-osteoblasts;Western blot was used to detect the relative expression of α-S protein. Results: When the concentration of DOPAL was ≥0.1 μmol·L^(-1), it had a significant inhibitory effect on the proliferation of MC3 T3-E1 pre-osteoblasts, and compared with the control group, the difference was statistically significant(P<0.05 or P<0.01). Compared with that in the control group, the relative expression of osteoblast marker factor mRNA in each concentration group of DOPAL was significantly down-regulated(P<0.05 or P<0.01), the relative expression of apoptosis marker factor cleaved-caspase-3 protein, the rate of apoptosis and the relative expression of α-S protein were significantly increased(P<0.01);there were statistically significant differences in the above indicators between the two concentration groups of DOPAL(0.1 μmol·L^(-1)and 0.5 μmol·L^(-1))(P<0.05 or P<0.01). In the control group without osteoinductive agent, no mineralized nodules appeared;compared with the osteoinduction group, DOPAL 0.1 μmol·L^(-1)and 0.5 μmol·L^(-1)groups inhibited the differentiation and maturation of MC3 T3-E1 cells. Conclusion: DOPAL inhibits the proliferation, differentiation and mineralization of pre-osteoblasts, and promotes the apoptosis of pre-osteoblasts, which may be

关 键 词：3 4-二羟基苯乙醛 前成骨细胞 骨质疏松 帕金森病 Α-突触核蛋白 

分 类 号：R965.1[医药卫生—药理学]