Ge Kui, Niu Yi-wen, Xie Ting, Lin Wei-dong, Tian Ming, Xu Bing, Cui Shi-tao, Lu Shu-liang
Department of Emergency, Tenth People's Hospital of Tongji University, Shanghai 200072, China.
Zhonghua Shao Shang Za Zhi. 2009 Dec;25(6):433-6.
To understand the influence of accumulation of advanced glycosylation end products (AGE) on wound healing of burn rats complicated with diabetes.
Seventy-five SD rats were divided into control, diabetes, and aminoguanidine-interfered groups in completely randomized method, with 25 rats in each group. All rats were subjected to deep partial-thickness scald. Diabetes was reproduced in rats of diabetes and aminoguanidine-interfered groups. Rats in aminoguanidine-interfered group were fed with 100 mg x kg(-1) xd (-1) aminoguanidine. Rats were sacrificed on post-scald day (PSD) 0, 3, 7, 14, and 21, and portrait of the wounds were taken. Full-thickness skin tissue specimens were obtained for determination. Specimens of epidermis from back of SD rats were obtained for KC cultivation and verification. Wound healing rate, glucose content in skin tissue, morphologic change in wound tissue, AGE distribution in skin tissue, influence of AGE on proliferation and apoptosis of KC were observed.
Wound healing rate of rats was respectively lower in diabetes group than that in control group on PSD 7, 14, and 21 (P < 0.01), but it was obviously higher in aminoguanidine-interfered group than that in the former 2 groups (P < 0.01). Glucose content of rat skin in diabetes group was (2.62 +/- 0.19) mmol/g, and it was (2.58 +/- 0.07) mmol/g in aminoguanidine-interfered group, both higher than that in control group [(1.04 +/- 0.09) mmol/g, P < 0.01]. In control group, limited intensive infiltration of inflammatory cells was found in the wound with necrotic tissue formation which fell off in time, and with no obvious delay of wound healing. In diabetes group, infiltration of inflammatory cells in wounds of rats appeared slowly, but diffusely and persistently; necrotic tissue formed and fell off late in time, with obvious delay of wound healing. In aminoguanidine-interfered group, intensive infiltration of inflammatory cells was observed in time, and the time of necrotic tissue formation and sloughing, and wound healing were respectively earlier than that in diabetes group. Sporadic disposition of small amount of AGE was found in rats in control group. AGE accumulation increased significantly in rats in diabetes group. AGE content decreased significantly in rats in aminoguanidine-interfered group after administration of aminoguanidine. KC proliferation decreased significantly in concentration dependent manner 48 hours after AGE stimulation. Absorbance value of AGE decreased in each AGE-interfered group (P < 0.01). Early Annexin-V positive apoptotic KC rate was obviously higher in 100 ug/mL AGE-interfered group (15.1 +/- 2.3)% than that in control group [(11.2 +/- 1.2)%, P < 0.05]. There was no statistical significance between 100 ug/mL AGE-interfered group (14.3 +/- 3.5)% and control group (15.2 +/- 2.4)% in respect of the rate of double-positive cells apoptosis at final stage (P > 0.05).
Hyperglycemia may inhibit proliferation of repairing cells such as KC through AGE accumulation, thus impedes wound healing. Reduction of AGE accumulation could ameliorate wound healing delay due to diabetes.
了解晚期糖基化终末产物（AGE）蓄积对糖尿病合并烧伤大鼠创面愈合的影响。
75只SD大鼠采用完全随机法分为对照组、糖尿病组和氨基胍干预组，每组25只。所有大鼠均行深Ⅱ度烫伤。糖尿病组和氨基胍干预组大鼠制备糖尿病模型。氨基胍干预组大鼠给予100 mg·kg⁻¹·d⁻¹氨基胍灌胃。于烫伤后（PSD）0、3、7、14和21天处死大鼠，拍摄创面照片。获取全层皮肤组织标本进行检测。取SD大鼠背部表皮标本进行角质形成细胞（KC）培养及鉴定。观察创面愈合率、皮肤组织葡萄糖含量、创面组织形态学变化、皮肤组织中AGE分布、AGE对KC增殖及凋亡的影响。
糖尿病组大鼠在PSD 7、14和21天时创面愈合率分别低于对照组（P<0.01），但氨基胍干预组明显高于前两组（P<0.01）。糖尿病组大鼠皮肤葡萄糖含量为（2.62±0.19）mmol/g，氨基胍干预组为（2.58±0.07）mmol/g，均高于对照组[（1.04±0.09）mmol/g，P<0.01]。对照组创面有少量炎性细胞浸润，有坏死组织形成且及时脱落，创面愈合无明显延迟。糖尿病组大鼠创面炎性细胞浸润出现缓慢，但弥漫且持久；坏死组织形成及脱落时间晚，创面愈合明显延迟。氨基胍干预组炎性细胞及时大量浸润，坏死组织形成及脱落时间、创面愈合时间均早于糖尿病组。对照组大鼠可见少量AGE散在分布。糖尿病组大鼠AGE蓄积明显增加。氨基胍干预组大鼠给予氨基胍后AGE含量明显降低。AGE刺激48小时后，KC增殖呈浓度依赖性显著降低。各AGE干预组吸光度值降低（P<0.01）。早期膜联蛋白V阳性凋亡KC率在100μg/mL AGE干预组（15.1±2.3）%明显高于对照组[（11.2±1.2）%，P<0.05]。100μg/mL AGE干预组（14.3±3.5）%与对照组（15.2±2.4）%在终末期双阳性细胞凋亡率方面无统计学意义（P>0.05）。
高血糖可能通过AGE蓄积抑制KC等修复细胞的增殖，从而阻碍创面愈合。减少AGE蓄积可改善糖尿病所致的创面愈合延迟。
