瘦素对体外大鼠成纤维细胞增殖与胶原合成的影响

目的　研究瘦素对体外培养大鼠成纤维细胞 (fibroblast,FB)增殖与胶原合成的影响 ,以阐明 FB介导瘦素在大鼠皮肤创伤愈合中的促进作用。　方法　体外培养乳鼠真皮 FB,通过 MTT比色分析法、3H-胸腺嘧啶核苷(3H- Td R)和 3H-脯氨酸 (3H- Pro)掺入法观察不同浓度的瘦素 ,分别为 0、10、5 0、10 0、2 0 0及 4 0 0 ng/ ml对其增殖和胶原合成的影响。　结果　瘦素剂量在 4 0 0 ng/ ml以下时 ,随瘦素剂量增加明显增加了体外培养大鼠 FB MTT的光密度值和 3H- Td R的掺入量。3H- Pro的掺入量随瘦素剂量增加基本呈增大趋势。 2 0 0、4 0 0 ng/ ml剂量组的3H- Td R掺入量 379±10 1cpm、32 6± 33cpm,与对照组 2 19± 5 6 cpm比较有统计学差异 (P<0 .0 5 ) ;且 2 0 0、4 0 0 ng/ ml剂量组的 MTT吸光度(A)值 0 .0 82± 0 .0 13、0 .0 91± 0 .0 18与对照组 0 .0 6 3± 0 .0 10比较有统计学差异 (P<0 .0 5 ) ;2 0 0、4 0 0 ng/ ml剂量组的 3H- Pro掺入量 911± 5 5 cpm、10 72± 2 5 9cpm,与对照组 6 79± 176 cpm比较有统计学差异 (P<0 .0 5 )。　结论　瘦素促进了 FB的增殖和胶原蛋白的合成 ,这可能是瘦素发挥促进大鼠皮肤愈合作用的途径之一。

STUDY ON THE EFFECT OF LEPTIN ON FIBROBLAST PROLIFERATION AND COLLAGEN SYNTHESIS IN VITRO IN RATS

OBJECTIVE: To investigate the effect of leptin on fibroblast proliferation and collagen synthesis as to elucidate that fibroblasts play a role in leptin's effect on wound healing. METHODS: Purified dermal fibroblasts were derived from sucking wistar rat skin and exposed to leptin at concentration of 0, 10, 50, 100, 200, and 400 ng/ml. The survived fibroblasts were assessed by the colorimetric thiazolyl blue (MTT) assay. Replication of fibroblast was quantified by the incorporation of 3H-thymidine. Collagen synthesis of fibroblast cell was measured by the incorporation of 3H-proline into collagenase-sensitive protein. RESULTS: The absorption of fibroblast exposed to leptin at concentration of 200 and 400 ng/ml 0.082 +/- 0.013, 0.091 +/- 0.018 was higher than that of control group 0.063 +/- 0.010, P < 0.05. The incorporations of 3H-thymidine of fibroblast exposed to leptin at concentration of 200 and 400 ng/ml 379 +/- 101 cpm, 326 +/- 33 cpm were significantly higher than those of control group 219 +/- 56 cpm, P < 0.05. The incorporations of 3H-proline of fibroblast exposed to leptin at concentration of 200 and 400 ng/ml 911 +/- 55 cpm, 1 072 +/- 259 cpm were significantly higher than that of control group 679 +/- 176 cpm, P < 0.05. CONCLUSION: Lepin can promote rat cutaneous fibroblast proliferation and collagen synthesis in vitro. This suggests that cutaneous fibroblast plays a role in leptin's promoting skin wound healing and it may be one of the main mechanisms by which leptin enhances skin wound healing.