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[目的]探讨镉染毒和染毒中止后成骨细胞增殖、分化及矿化等生物学功能的改变。[方法]采用混合酶消化法,分离SD大鼠颅盖骨成骨细胞,在5%CO_2、37℃条件下培养24h后,持续镉染毒组以不同浓度的氯化镉(0～2.000μmol/L)持续染毒72h;间断镉染毒组以同样浓度的氯化镉染毒48h后,更换成无镉培养液继续培养24h,以噻唑蓝(MTT)法测定细胞增殖能力改变,用对硝基苯磷酸二钠盐(PNPP)偶氮法检测碱性磷酸酶(ALP)活性。同时,成骨细胞培养7d后,以0.500μmol/L氯化镉持续作用3～13d,然后分别停止作用10、8、6、0d,采用茜素红S进行矿化结节染色并计算面积以观察镉暴露中止后成骨细胞矿化能力损伤的恢复情况。[结果]氯化镉可明显抑制成骨细胞增殖、ALP活性及矿化能力。间断镉染毒组在停止作用24 h后,成骨细胞的增殖、分化仍然明显低于对照组(P<0.05),与持续作用组比较没有明显改善。同样,间断镉染毒组在停止作用不同时间后,成骨细胞矿化结节数量和面积仍然明显低于对照组(P<0.05),与持续作用组相比没有明显恢复。[结论]镉暴露中止一定时间后,镉对成骨细胞的影响作用仍明显存在,表现为对成骨细胞增殖、分化及矿化能力的持续抑制。
[Objective] To explore the changes of biological functions such as proliferation, differentiation and mineralization of osteoblasts after cadmium exposure and abortion. [Method] The calvarial osteoblasts of SD rats were isolated by mixed enzyme digestion and cultured in 5% CO 2, 37 ℃ for 24 h. The cadmium chloride (0 ~ 2.000 μmol / L) for 72h. The intermittent cadmium exposure group was treated with the same concentration of cadmium chloride for 48h, then replaced with cadmium-free medium for 24 h. The proliferation of cells was determined by MTT assay. Nitrophenyl phosphate disodium salt (PNPP) was used to detect alkaline phosphatase (ALP) activity. At the same time, osteoblasts were treated with 0.500μmol / L cadmium chloride for 3 ~ 13d, and then stopped for 10, 8, 6, and 0d respectively. Alizarin Red S was used to stain the mineralized nodules and the area To observe the recovery of mineralization ability of osteoblasts after cadmium exposure. [Results] Cadmium chloride could significantly inhibit osteoblast proliferation, ALP activity and mineralization ability. After discontinued for 24 h, the proliferation and differentiation of osteoblasts in the intermittent cadmium-exposed group were still significantly lower than those in the control group (P <0.05), but no significant improvement compared with the continuous group. Similarly, the number and area of mineralized nodules of osteoblasts after discontinuation of cadmium exposure were still significantly lower than those of the control group (P <0.05), and no significant recovery compared with the sustained-effect group. [Conclusion] The effect of cadmium on osteoblasts is still obvious after the cadmium exposure is suspended for a certain period of time, which shows the continuous inhibition on the proliferation, differentiation and mineralization of osteoblasts.