透明质酸钠介导的创面修复中真皮细胞增殖周期的动态变化

Dynamic changes of dermal cell proliferation cycle in the repair of wound mediated by sodium hyaluronate

目的本研究利用透明质酸钠作为磺胺嘧啶银药物载体作用于大鼠烧伤创面,观察大鼠烧伤创面真皮细胞周期DNA分布情况,以期制备出一种加快速面愈合的新型外用制剂。方法自制恒温恒压烧伤仪复制SD大鼠Ⅱ~Ⅲ度烧伤模型,创面标记后分别外涂安慰剂、磺胺嘧啶银混悬剂、透明质酸钠、磺胺嘧啶银和透明质酸钠混合制剂,行暴露疗法,创面每2 d清创后涂药1次,连续28 d,分别于烧伤后第3、5、7、10、14、21天采用颈椎脱臼法分批处死SD大鼠,每批5只。观察烧伤创面渗出和愈合情况,行创面皮肤组织病理和生化检测,机械加胰酶法制备标本,采用流式细胞仪测定烧伤后创面真皮细胞G0/G1期和S期DNA含量。结果烧伤后第3、5、7、10、14、21天,磺胺嘧啶银组、透明质酸钠组和混合制剂组大鼠真皮细胞G0/G1期DNA含量均显著低于同期空白组(均P<0.05);烧伤后第5天和第7天,透明质酸钠组大鼠真皮细胞G0/G1期DNA含量均显著低于同期磺胺嘧啶银组(均P<0.05);烧伤后第3天和第5天,混合制剂组大鼠真皮细胞G0/G1期DNA含量均显著高于同期磺胺嘧啶银组和透明质酸钠组(均P<0.05),烧伤后第7、10、14、21天混合制剂组大鼠真皮细胞G0/G1期DNA含量均显著低于同期磺胺嘧啶银组和透明质酸钠组(均P<0.05)。烧伤后第3、5、7、10天,磺胺嘧啶银组大鼠真皮细胞S期DNA含量均显著高于同期空白组(均P<0.05);烧伤后第3、5、7、10、14、21天,透明质酸钠组和混合制剂组大鼠真皮细胞S期DNA含量均显著高于同期空白组(均P<0.05);烧伤后第5、14、21天,透明质酸钠组大鼠真皮细胞S期DNA含量均显著高于同期磺胺嘧啶银组(均P<0.05);烧伤后第7、10、14、21天,混合制剂组大鼠真皮细胞S期DNA含量均显著高于同期磺胺嘧啶银组和透明质酸钠组(均P<0.05)。结论透明质酸钠可促使大量处于G1和G0期的静止细胞进入S期进行DNA复制,细胞增殖效果明显。

Objective This study used sodium hyaluronic as a drug carrier compound with sulfadiazine silver,observed the DNA distribution of dermal cell cycle,in order to prepare of a new topical preparation to accelerate wound healing.Method SD rats'burn models of degreeⅡ-Ⅲwere reproduced by a self-made constant temperature and pressure burn apparatus.After the wound was marked,the mixture of placebo,silver sulfadiazine suspension,sodium hyaluronate,mixture of silver sulfadiazine and sodium hyaluronate were applied to the wound surface for exposure therapy.The wounds were cleaned and coated once every two days for 28 days.SD rats'were killed in batches by cervical dislocation on the 3rd,5th,7th,10th,14th and 21st day after burned,5 rats in each batch.Observed the exudation and healing of burn wounds,detected the histopathology and biochemistry of the wounds,prepared specimens by mechanical method and trypsin method,and measured the DNA content of the dermal cells at G0/G1 and S stages after the wounds by flow cytometry.Result On the 3rd,5th,7th,10th,14th and 21st day after burned,the contents of DNA in G0/G1 phase of dermal cells in silver sulfadiazine group,sodium hyaluronate group and mixed preparation group were significantly lower than those in blank group(all P<0.05);on the 5th and 7th day after burned,the contents of DNA in G0/G1 phase of dermal cells in sodium hyaluronate group were significantly lower than those in silver sulfadiazine group(all P<0.05);on the 3rd and 5th day after burned,the contents of DNA in mixed preparation group were significantly higher than those in silver sulfadiazine group and sodium hyaluronate group(all P<0.05),the contents of DNA in G0/G1 phase of dermal cells on the 7th,10th,14th and 21st day after burned were significantly lower than those in silver sulfadiazine group and sodium hyaluronate group(all P<0.05).On the 3rd,5th,7th and 10th day after burned,the contents of S-phase DNA in dermal cells of silver sulfadiazine group were significantly higher than those of blank group(all P<0.05);on the 3rd,5th,7th,10th,14th and 21st day after burned,the contents of S-phase DNA in dermal cells of sodium hyaluronate group and mixed preparation group were significantly higher than those of blank group(all P<0.05);on the 5th,14th and 21st day after burned,the contents of S-phase DNA in dermal cells of sodium hyaluronate group were significantly higher than those of silver sulfadiazine group(all P<0.05);on the 7th,10th,14th and 21st day after burned,the contents of DNA in S-phase of dermal cells in the mixed preparation group were significantly higher than those in silver sulfadiazine group and sodium hyaluronate group(all P<0.05).Conclusion Sodium hyaluronate can promote a large number of stationary cells in G1 and G0 phase to enter S phase,and the effect of DNA replication cell proliferation is obviously.