Rho激酶抑制剂Y-27632对人牙髓干细胞增殖能力的影响

目的: 初步研究Rho激酶抑制剂Y-27632对人牙髓干细胞(human dental pulp stem cells,hDPSCs)增殖能力的影响。方法: 采用体外贴壁式培养法培养人牙髓干细胞,应用Rho激酶抑制剂Y-27632,根据培养条件,分为普通培养液培养组(Con)及普通培养液+抑制剂Y-27632培养组(Con+Y),采用MTT法检测两种培养条件下人牙髓干细胞培养24、48、72、96 h细胞活性;采用DAPI染色法及流式细胞定量检测技术(FCM)比较两组在培养48 h的细胞数量及细胞周期分布情况。结果: MTT结果显示,实验组hDPSCs细胞增殖曲线较对照组明显上移,且增殖高峰期提前;培养48 h,DAPI染色结果显示,实验组细胞总数量明显多于对照组;FCM结果显示,对照组G₀/G₁期细胞比例为(66.8±6.84)%,S期细胞比例为(25.17±0.62)%,实验组G0/G1期细胞比例为(58.59±1.76)%,S期细胞比例为(31.34±1.16)%,实验组S期细胞比例明显高于对照组(P<0.05)。 结论: Rho激酶抑制剂Y-27632促进人牙髓干细胞DNA合成、细胞分裂及增殖。

Functional Role of Rho Kinase Inhibitor Y-27632 on Proliferation of Human Dental Pulp Stem Cells.

Objective: To study the influence of Rho kinase inhibitor Y-27632 on the proliferation of human dental pulp stem cells. Methods: Human dental pulp stem cells (hDPSCs) were seeded in plates，then divided randomly into two groups: cultured with or without Y-27632. Cellular proliferation was determined by DAPI staining and MTT assay. Cell cycle was analyzed by flow cytometry (FCM). Results: After 24, 48, 72 and 96h of cultivation, the optical density (OD) in the Y-27632 treated group was significantly higher than that in the control group. Moreover, the peak time during proliferation was earlier in the treat group. After 48h cultured, compared with the control group, the DAPI staining showed that the proportion of total cells was significantly increased in the Y-27632 treated group. The FCM results indicated that (66.8±6.84)% of the hDPSCs were in G0/G1 phase (25.17±0.62)% in S phase. After Y-27632 treatment, (58.59±1.76)% of the hDPSCs were in G0/G1 phase (31.34±1.16)% in S phase. The cell proliferation during S phase was significantly higher after stimulation by Y-27632 than that in the control group (P<0.05). Conclusion: Inactivation of Rho kinase in the hDPSCs by inhibitor Y-27632 led more hDPSCs to entering into the DNA synthesis stage and promoted the division and proliferation of hDPSCs.