特异性整合素连接激酶抑制剂QLT0267对大鼠烧伤创面愈合影响的初步研究

目的 观察局部注射特异性整合素连接激酶（ILK）抑制剂QLT0267对SD大鼠深Ⅱ度烧伤创面愈合影响,初步探讨ILK在创面愈合过程中的作用及机制.方法 选取45只雄性SD大鼠,采用恒温恒压烫伤仪在其背部皮肤制备深Ⅱ度烧伤创面,并随机分成实验组、对照组及二甲基亚砜（DMSO）组,每组15只.实验组创面每天注射浓度100 μM QLT0267液100 μL;对照组创面每天注射0.9％氯化钠溶液100 μL;DMSO组创面每天注射0.3％ DMSO液100 μL.测量各组大鼠创面愈合时间和愈合率;伤后14 d取材,各组随机选择5只大鼠,用SP法检测ILK、AKT、PAKT、α-SMA在创面组织中的表达,Western Blot检测各组ILK、AKT、PAKT蛋白含量;用Masson改良法检测创面胶原.结果 实验组大鼠创面平均愈合时间为（21.1±0.6）d,比对照组（17.1±0.6）d和DMSO组（17.7 ±0.6）d长;实验组创面愈合率也低于对照组和DMSO组,比较差异有统计学意义（P＜0.05）.Western Blot检测结果显示：各组ILK、AKT蛋白表达差异无统计学意义（P＞0.05）,实验组PAKT蛋白显著低于对照组和DMSO组.SP法结果各组ILK、AKT表达差异无统计学意义（P＞0.05）,实验组ILK活性被抑制,PAKT（0.406±0.008）表达较对照组（0.901 ±0.013）、DMSO组（0.966±0.011）降低,比较差异有统计学意义（F=11.27,P=0.04）;实验组α-SMA表达（0.201±0.003）较对照组（0.339 ±0.006）、DMSO组（0.351 ±0.005）也降低,比较差异有统计学意义（F=52.86,P=0.001）;实验组细胞外基质胶原排列较对照组、DMSO组明显稀疏、紊乱.结论 ILK活性被抑制时延迟大鼠皮肤创面愈合.ILK特异性抑制剂QLT0267通过抑制ILK活性PAKT合成减少,可能影响其下游信号的传导,导致肌成纤维细胞生成减少、胶原合成减少,影响创面愈合.

Effect of integrin-linked kinase specific inhibitor QLT0267 on burn wound healing in rats

Objective To observe the effect of specific inhibitor for integrin-linked kinase（ILK） on wound healing of deep partial-thickness burns in rats and explore the mechanism of ILK in wound healing process. Methods Deep partial-thickness burns models were created with a scald constant temperature and pressure in its back in 45 male SD rats and then they were divided into the experimental group, control group and DMSO group randomly, 15 rats in each group. The experimental group daily injection concentrations of 100 μM QLT0267 solution at 100 μL, the control group was injected daily for 0. 9% sodium chloride solution at 100 μL, DMSO group was injected daily with 0.3% DMSO solution at 100 μL. Wound healing rate were measured and recorded everyday. The expressions of ILK, AKT, PAKT and α-SMA in wound tissue were investigated with SP method 14 days after burn. The proteins of ILK, AKT and PAKT in each group were investigated with Western Blot. Collagen in wounds was determined with Masson method. Results The period of wound healing time in experimental group （21.1 ± 0.64） d was significantly longer than that in control group （17.1 ±0.6） d and in DMSO group （17.7 ± 0.6）d. There was no significant difference in expressions of ILK and AKT with Strepeavidin-Perosidase and Western Blot in various groups. But the expression of PAKT（0. 406 ± 0. 008 ） in experimental group was significant less than that in control group （0. 901 ± 0.013） and DMSO group （0. 966 ± 0.011 ） （ F = 11.27, P = 0.04 ）. The expression of α- SMA significantly decreased in experimental group （0. 201 ±0.003）than that in Control group （0. 339 ± 0.006） and DMSO group （ 0.351 ± 0. 005 ） （ F = 52. 86, P = 0.001 ）. The experimental group of extracellular matrix collagen compared with the control group, DMSO group was sparse disordered. Conclusions ILK inhibitor QLT0267 may affect ILK＇s transmission of its downstream signals by inhibiting ILK activity and PAKT generation, which can result in limitatio