热休克蛋白60诱导小鼠皮肤移植耐受的实验研究  

EXPERIMENTAL STUDY ON INDUCTION OF SKIN ALLOGRAFT TOLERANCE IN MICE BY HEAT SHOCK PROTEIN 60

作　　者：黄文鹏[1] 叶启发[1] 黄祖发[1] 杨定华[2] 彭民浩[2] 

机构地区：[1]中南大学移植医学研究院卫生部移植医学工程技术研究中心中南大学湘雅三医院,长沙410013 [2]广西医科大学第一附属医院肝胆外科

出　　处：《中国修复重建外科杂志》2008年第8期993-997,共5页Chinese Journal of Reparative and Reconstructive Surgery

摘　　要：目的探讨热休克蛋白60(heat shock protein 60,HSP60)对小鼠移植皮片成活的影响及其机制。方法选用60只C57BL/6(H-2b)小鼠为受体,45只BALB/C(H-2d)小鼠、15只CBA/N(H-2k)为供体,均为8～12周龄近交系雌性小鼠。受体小鼠剪下1cm×1cm全层皮肤,干纱布清创,即为植床;随机分为4组,每组15只。A组:无菌取供体BALB/C(H-2d)小鼠背部1cm×1cm全层皮片,刮去皮下组织后移植至受体小鼠背部;B组:受体小鼠背部皮下注射0.1mL不完全弗氏佐剂(imcompleted Freund’s adjuvant,IFA),2周后行BALB/C(H-2d)小鼠皮肤移植;C组:受体小鼠背部皮下注射经0.1mLIFA乳化后的50μgHSP60,2周后行BALB/C(H-2d)小鼠皮肤移植;D组:受体小鼠背部皮下注射经0.1mLIFA乳化后的50μgHSP60,2周后行CBA/N(H-2k)小鼠皮肤移植。B、C、D组供体皮肤移植方法同A组。术后观察移植皮片成活时间;移植术后7、25d行单向混合淋巴细胞反应及混合淋巴细胞培养上清液细胞因子检测;术后7d行迟发型超敏反应测定。结果A、B、C、D组移植皮片成活时间分别为(12.4±0.5)、(11.6±0.8)、(29.3±2.6)及(27.6±2.1)d;A、B组与C、D组比较,差异有统计学意义(P<0.05),A、B组间及C、D组间比较,差异无统计学意义(P>0.05)。皮肤移植术后7d,A、B、C、D组混合淋巴细胞反应每分钟放射脉冲数(counts of perminute impulse,cmp)值分别为12836±1357、11876±1265、6581±573及6843±612;A、B组与C、D组比较,差异有统计学意义(P<0.05);A、B组间及C、D组间比较差异无统计学意义(P>0.05);术后25d,各组cpm值分别为13286±1498、12960±1376、11936±1265及12374±1269,各组间差异无统计学意义(P>0.05)。皮肤移植术后7d,C、D组混合淋巴细胞培养上清液IL-10高于A、B组,IL-2、干扰素γ(interferon γ,IFN-γ)低于A、B组(P<0.05),A、B组间及C、D组间差异均无统计学意义(P>0.05);术后25d,各组IL-2、IL-10及IFN-γ差异无统计学意义(P>0.05)。皮肤移植术后7d,A、B、C、D组受体小�Objective To investigate the role and mechanism of heat shock protein 60 （HSP60） in induction of murine skin allograft tolerance. Methods At the age of 8-12 weeks, inbred female BALB/C （H-2d） mice （n=45） and CBA/N （H-2k）mice （n=15） were used as transplantation donors and C57BL/6 （H-2b） mice （n=60） as recipients. Recipients C57BL/6 （H-2b） mice were randomized into 4 groups （n=15）. In group A, 1 cm × 1 cm Wolfe-Krause skin graft was excised from the back of BALB/C （H-2d） mice and hypoderma was scraped off aseptically, and then transplanted to the back of C57BL/6 （H-2b） mice. The method of skin transplantation in the other 3 groups was the same as to group A. In group B, C57BL/6 （H-2b） mice were treated with imcompleted Freund＇s adjuvant （IFA） administration into the back 2 weeks before transplantation of BALB/C （H-2^d） mice skin. In group C, C57BL/6 （H-2^b） mice were administered HSP60 emulsified in IFA into the back 2 weeks before transplantation of BALB/C （H-2 ^d） mice skin. In group D, C57BL/6 （H-2^b） mice were treated with HSP60 emulsified in IFA into the back and followed by skin transplantation of CBA/N （H-2^k） mice 2 weeks later. The delayed type hypersensitivity was determined 7 days after transplantation. One-way mixed lymphocyte reaction, the concentration of cytokines in the mixedlymphocyte reaction culture supernatant was determined 7 days and 25 days after transplantation. The survival time of skin allograft was observed. Results The survival time of skin allograft in groups A, B, C and D was 12.4 ±0.5, 11.6 ± 0.8, 29.3 ± 2.6 and 27.6 ± 2.1 days, respectively. There was significant difference between groups A, B and groups C, D （P 〈 0.05）, while there was no significant difference between group A and group B as well as between group C and group D （P 〉 0.05）. The counts of per minute impulse （cpm） of mixed lymphocyte reaction 7 days after transplantation in groups A, B, C and D was 12 836 ± 1357, 11876 ± 126

关 键 词：热休克蛋白60 皮肤移植 免疫耐受 小鼠 

分 类 号：R622.1[医药卫生—整形外科] R392.4[医药卫生—外科学]