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目的摸索体外培养体系中不影响人脐静脉平滑肌细胞(HUVSMCs)生长的硫氢化钠(NaHS)适宜浓度。方法采用原代培养人脐静脉平滑肌细胞,实验分为八组,分别是正常对照组和七个NaHS浓度组,依次为:5.0×10-3M、1.0×10-3M、2.0×10-4M、4.0×10-5M、8.0×10-6M、1.6×10-6M、3.2×10-7M。采用细胞计数法绘制细胞生长曲线并观察细胞的生长状态,MTT检测细胞生长活力,流式细胞术检测细胞周期并进行比较。结果 4.0×10-5M组、8.0×10-6M组、1.6×10-6M组、3.2×10-7M组从细胞生长活力、细胞周期及细胞生长曲线绘制三方面与对照组相比,差异均无统计学意义(P>0.05);5.0×10-3M组、1.0×10-3M组、2.0×10-4M组与对照组相比,细胞生长受到抑制,差异有统计学意义(P<0.05)。结论 1.6×10-6M～4.0×10-5M中任意浓度的NaHS均不影响HUVSMCs的正常生长,其可作为培养HUVSMCs并作为实验材料的浓度范围。
Objective To explore the appropriate sodium hydrosulfide (NaHS) concentration in human in vitro culture system that does not affect the growth of human umbilical vein smooth muscle cells (HUVSMCs). Methods Primary cultured human umbilical vein smooth muscle cells were divided into eight groups: normal control group and seven NaHS concentration groups, which were 5.0 × 10-3M, 1.0 × 10-3M, 2.0 × 10-4M, 4.0 × 10 -5 M, 8.0 × 10 -6 M, 1.6 × 10 -6 M, 3.2 × 10 -7 M. Cell growth curve was drawn by cell counting method and cell growth was observed. Cell viability was measured by MTT assay. Cell cycle was detected by flow cytometry and compared. Results Compared with the control group, the cell viability, cell cycle and cell growth curve were significantly different in 4.0 × 10-5M, 8.0 × 10-6M, 1.6 × 10-6M and 3.2 × 10-7M groups (P> 0.05). Compared with the control group, the cell growth was inhibited in 5.0 × 10-3M group, 1.0 × 10-3M group and 2.0 × 10-4M group, the difference was statistically significant (P <0.05 ). Conclusion Any concentration of NaHS from 1.6 × 10-6 M to 4.0 × 10-5 M did not affect the normal growth of HUVSMCs, which could be used as the concentration range of HUVSMCs.