粉防己碱对瘢痕成纤维细胞活性功能的影响

Influence of tetrandrine on viability of scar derived fibroblasts in vitro

目的 观察粉防己碱对体外培养的巨噬细胞介质介导瘢痕成纤维细胞活性功能的调控作用 ,并探讨其在瘢痕防治中的意义。方法 取 5例手术切除的人增生性瘢痕组织标本行瘢痕成纤维细胞体外培养 ,于传代培养至第 4代的成纤维细胞 (5× 10 4)中分组加入巨噬细胞介质、粉防己碱继续培养 ,然后分别用3 H TdR和3 H 脯氨酸脉冲标记 ,放射性核素液闪烁仪定量检测各组3 H TdR和3 H 脯氨酸掺入值。并采用三维培养方法测定体外模拟瘢痕样组织的收缩指数变化。结果 粉防己碱 3mg/L不抑制细胞DNA合成时 ,即可明显减少瘢痕成纤维细胞 3 H 脯氨酸掺入值 ,由对照组的 94 5± 191减少至6 82± 2 0 5 (P <0 .0 5 ) ;并能显著降低巨噬细胞介质介导的瘢痕成纤维细胞3 H TdR掺入值和3 H 脯氨酸掺入值 ,由单纯介质组的 2 75 6± 2 4 4、1384± 2 93分别降低至 194 8± 2 82、10 34± 14 2 (均P <0 .0 1)。粉防己碱 1mg/L即可降低体外瘢痕样组织的收缩指数 (P <0 .0 1)。结论 粉防己碱对体外培养的巨噬细胞介质介导瘢痕成纤维细胞增殖、胶原合成以及收缩三方面具有抑制效应 ,可望在增生性瘢痕的防治中发挥重要作用。

Objective To explore the regulative effects of tetrandrine on viability of scar derived fibroblasts stimulated by the inflammation mediators released by activated macrophages and to evaluate the role of tetrandrine in prevention and treatment of scar. Methods The fibroblasts were isolated from five cases of human hypertrophic scar tissues for culture in vitro. The cell cultures at passages 4 were used in this experiment. Tetrandrine and inflammation mediators released by activated macrophages were added to culture medium respectively. Then the cultures were palse labeled with 3H TdR and 3H Proline. TdR and proline uptake into fibroblasts were determined quantitatively. And the contraction indexes of scar tissue like structure in vitro were measured by using three dimensional culture system. Results When tetrandrine was at a concentration of 3mg/L, 3H TdR incorporation showed no statistical difference ( P >0.05 ). However, the absorption of 3H Proline were lower in experimental group 682±205 than those in control group 945±191( P <0.05). And also tetrandrine could obviously decreased the incorporation rates of 3H TdR and 3H Proline of scar derived fibroblasts mediated by inflammation mediators, which declined from 2756±244 and 1384±293 in mediator group to 1 948±282 and 1 034±142 in treatment group respectively ( P <0.01 ). Even at a concentration of 1 mg/L, the contraction indexes of scar tissue like structure were decreased significantly in experimental group compared to the control group ( P < 0.01 ) . Conclusion Tetrandrine can inhibit not only the proliferation and collagen synthesis of scar derived fibroblasts stimulated by inflammation mediators released by activated macrophages, but also the contraction of scar tissue like structure in vitro. It suggests that tetrandrine may play an important role in the prevention and treatment of hypertrophic scars.