利多卡因通过增加细胞周期素依赖的蛋白激酶抑制剂1A（p21）表达抑制NIH-3T3细胞的增殖

背景研究利多卡因抑制大鼠胚胎成纤维细胞株NIH-3T3增殖的分子机制。体外实验中，局麻药可抑制细胞增殖，但它们对伤口愈合的影响目前尚存在争议。我们观察了利多卡因对体外大鼠成纤维细胞株NIH一3T3增殖的作用及机制。方法将NIH-3T3细胞置于含利多卡因（O、0．05、0．5、1、2、5rIM）的培养液中生长。细胞计数反映细胞增殖水平，并测定溴脱氧尿苷的摄取来反映利多卡因的抑制作用，基因表达和蛋白增殖水平分别采用PcR和westem测定。结果利多卡因呈浓度依赖性地抑制NIH—313细胞增殖，其抑制程度从无（Q05和Q5mM）到轻微（1mM）到很强（2mM和5mM）（P=0．006）。第3．5天，2mM利多卡因组明显抑制溴脱氧尿苷的摄取（与对照组相比P=0．02，与lmM利多卡因组相比P=Q0495）。第1．5天，利多卡因可上调细胞周期蛋白D1和细胞周期素依赖性蛋白激酶抑制剂1A（p21）的表达。第2．5天，利多卡因增加了p21蛋白水平。结论在脊髓麻醉、硬膜外麻醉和神经阻滞时，血浆中利多卡因浓度较低，其对成纤维细胞的增殖无明显影响，而高浓度的利多卡因可升高p21蛋白水平，使细胞增殖停止在细胞周期的S期，从而抑制细胞增殖。组织渗透后可能达到的高浓度利多卡因可能通过抑制成纤维细胞增殖而阻碍伤口的愈合。

BACKGROUND： We explored molecular mechanisms by which lidocaine inhibits growth in the murine em- bryonic fibroblast cell line NIH-3T3. Local anesthetics can adversely affect ceil growth in vitro. Their effects on wound healing are controversial. We examined the effects and novel mechanisms by which lidocaine affects in vitro multiplication of the mu- fine fibroblast cell line NIH-3T3. METHODS： NIH,3T3 cells were grown in culture with lidocaine [0, 0. 05, 0. 5, 1, 2, and 5 mM]. Cell multiplication was assessed by determining cell counts on subsequent days, while mechanisms by which inhibi- tion occurred were evaluated by bromodeoxyuridine uptake, gene expression using polymerase chain reaction array, and Western blot analysis to verify increased levels of affected proteins. RESULTS： Lidocaine caused dose-dependent inhibitionof multiplication of NIH-3T3 cells. Effects ranged from no inhibition [0.05 and 0. 5 mM] and mild inhibition k 1 mMJ, to severe inhibition [ 2 and 5 mM] [ P = 0.006 ]. Lidocaine 2 mM inhibited bromodeoxyuridine uptake at day 3.5 [ P = 0. 02 versus control, and P = 0. 0495 vs 1 mM lidocaine ]. On day 1.5, lidocaiile upregulated expression of cyclin-D1 and cydin- dependent kinase inhibitor 1A [ p21 ]. On day 2.5, lidocaine increased the levels of p21 protein. CONCLUSIONS： Low con- centrations of lidocaine, as would be seen in plasma after spinal, epidural, or plexus anesthesia, do not significantly affect mul- tiplication of fibroblasts. Higher doses of lidocaine arrest cell multiplication at the S-phase of the growth cyde by upregulation of p21, an extremely potent inhibitor of cell multiplication. Higher concentrations, as would be seen after tissue infiltration, severely inhibit fibroblast multiplication and thus may impair wound healing.