摘要
目的探讨双氢睾酮和羟基氟他胺(以下称为DH)最佳配比,构建雄激素及其拮抗剂双重释放系统,分析该系统在小鼠Ⅲ度烧伤创面修复中的应用效果。方法该研究为实验研究。取HaCaT细胞,按随机数字表法(分组方法下同)分为空白组(不加药物培养)、低基线组、中基线组、高基线组,对后3组细胞均分别加用3种配比的DH培养。中配比下,从低到高3个基线组中双氢睾酮质量依次为1.4、2.8、4.0µg,羟基氟他胺质量依次为1.2、1.6、2.0µg,以此为基准,小配比下,双氢睾酮质量减少1/2,羟基氟他胺质量增加1/2;大配比下,双氢睾酮质量增加1/2,羟基氟他胺质量减少1/2。培养2 d,采用细胞计数试剂盒8检测细胞增殖水平(样本数为4)。取16只6~8周龄雄性BALB/c小鼠,于其背部制备Ⅲ度烧伤创面后,分为空白组、小配比组、中配比组和大配比组(每组4只小鼠),伤后7 d于后3组小鼠创面局部滴加含不同配比DH的生理盐水[从小到大3个配比组中DH总量分别为127.5、165.0、202.5µg,双氢睾酮与羟基氟他胺质量比(以下称为药物质量比)分别为8∶9、8∶3、8∶1],之后每48小时重复给药1次,直至伤后27 d;对空白组小鼠创面于前述时间点滴加生理盐水。观察伤后0(即刻)、7、14、21、28 d创面愈合情况并计算伤后7、14、21、28 d创面愈合率(样本数为4);伤后28 d,取创面组织,行苏木精-伊红染色后观察再上皮化情况,行Masson染色后观察胶原纤维情况并分析胶原纤维占比(样本数为3)。取20只6~8周龄雄性BALB/c小鼠,于其背部制备Ⅲ度烧伤创面后,分为普通支架组和小比例支架组、中比例支架组、大比例支架组(每组5只小鼠),伤后7 d开始对创面分别持续外敷运用静电纺丝技术制备的不含药物的聚己内酯支架和含药物质量比为1∶3、1∶1、3∶1的DH的聚己内酯支架(即雄激素及其拮抗剂双重释放系统,DH总量均约为1.7 mg)直至实验结束。
Objective To explore the optimal ratio of dihydrotestosterone and hydroxyflutamide(hereinafter referred to as DH),construct a dual release system of androgen and its antagonist,and analyze the application effect of this system in the repair of full-thickness burn wounds in mice.Methods This study was an experimental study.The HaCaT cells were divided into blank group(without drug culture),low baseline group,medium baseline group,and high baseline group according to the random number table(the same grouping method below),and the last three groups of cells were cultured by adding three different ratios of DH.Under a medium ratio,the mass of dihydrotestosterone in the three baseline groups from low to high was 1.4,2.8,and 4.0µg,respectively,and the mass of hydroxyflutamide was 1.2,1.6,and 2.0µg,respectively.On this basis,under a small ratio,the mass of dihydrotestosterone was reduced by half and the mass of hydroxyflutamide was increased by half;under a large ratio,the mass of dihydrotestosterone was increased by half and the mass of hydroxyflutamide was reduced by half.After culture of 2 days,the cell proliferation level was detected by cell counting kit 8(n=4).Sixteen 6-8-week-old male BALB/c mice were used to establish a full-thickness burn wound on the back and divided into blank group,small ratio group,medium ratio group,and large ratio group,with 4 mice in each group.On post injury day(PID)7,normal saline containing different ratios of DH was locally dropped to the wounds of mice in the last three groups of mice(the total mass of DH in the three ratio groups from small to large was 127.5,165.0,and 202.5µg,respectively,and the mass ratios of dihydrotestosterone to hydroxyflutamide(hereinafter referred to as drug mass ratio)were 8∶9,8∶3,and 8∶1,respectively),afterwards,the administration was repeated every 48 hours until PID 27;normal saline was dropped to the wound of mice in blank group at the aforementioned time points.The wound healing status on PID 0(immediately),7,14,21,and 28 was observed,and the
作者
黄日中
王裔惟
黄何艳
蒋蕊韩
薛楠楠
尹少平
赵宏宇
Huang Rizhong;Wang Yiwei;Huang Heyan;Jiang Ruihan;Xue Nannan;Yin Shaoping;Zhao Hongyu(Jiangsu Provincial Research Center for Development and Application of External Medicine of Traditional Chinese Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China;Clinical Research Center,the Second Hospital of Nanjing,Nanjing University of Chinese Medicine,Nanjing 210003,China)
出处
《中华烧伤与创面修复杂志》
CAS
CSCD
北大核心
2024年第2期180-189,共10页
Chinese Journal of Burns And Wounds
基金
国家自然科学基金青年科学基金项目(82003305)
江苏省高等学校基础科学(自然科学)研究面上项目(21KJB360016)
南京中医药大学自然科学基金项目(XZR2020069)。
