摘要
目的观察外敷氨基胍霜剂对糖尿病大鼠皮肤组织晚期糖基化终末产物（AGE）形成、Kc细胞增殖及氧化应激的影响。方法将硬脂酸、液状石蜡、凡士林、羊毛脂、肉豆蔻酸异丙酯、甘油、50g／L尼泊金醇、盐酸氨基胍等试剂按一定比例混合制成氨基胍霜剂，以不含有氨基胍的霜剂为基质。取健康大鼠背部皮肤，分别用5、10g／L氨基胍霜剂和5g／L氨基胍＋10g／L氮酮霜剂处理，于用药后2、4、7、10、12、24h测定药物透皮效果。将30只SD大鼠按随机数字表法分为正常对照组6只、糖尿病组8只、氨基胍治疗组8只、基质治疗组8只。后3组大鼠腹腔注射链脲佐菌素65mg／kg，诱导糖尿病模型；对照组大鼠注射0．05mmol／L柠檬酸缓冲液。注射1周后，正常对照组与糖尿病组大鼠不行任何治疗，氨基胍治疗组与基质治疗组大鼠背部分别连续外用10g／L氨基胍霜剂与基质治疗4周。取各组皮肤组织，胶原提取液荧光强度检测法测定AGE含量，流式细胞仪分析表皮KC周期，检测氧化应激相关指标超氧化物歧化酶（SOD）、丙二醛、总抗氧化能力、髓过氧化物酶（MPO）含量。对实验数据行t检验。结果10g／L氨基胍霜剂透皮效果优于5g／L氨基胍和5g／L氨基胍＋10g／L氮酮的霜剂。1只基质治疗组大鼠未诱导成功。建模后4周，糖尿病组与氨基胍治疗组大鼠分别死亡4只和1只。糖尿病组大鼠皮肤组织AGE含量为每毫克羟脯氨酸（OHP）中（36．8±2．6）U，明显高于正常对照组的每毫克OHP中（24．6±2．7）u（t=7．2，P〈0．01）；氨基胍治疗组AGE含量为每毫克OHP中（28．6±3．7）u，明显低于糖尿病组（t=-3．9，P〈0．05）；基质治疗组AGE含量[每毫克OHP中（32．2±5．2）U]与糖尿病组相近（t=1．6，P〉0．05）。糖尿病组大鼠S期KC比例为（5．3±0．6）％，低于正常对照组的（7．6±0．9）％（t=4．50，P〈0．01）；氨基胍治疗组大鼠S期和G2／M期KC比例均明显高于糖尿病组（t值分别为6．80、3．17，P值均小于0．01）；基质治疗组大鼠S期KC比例[（9．2±1．5）％]显著高于糖尿病组（t=4．90，P〈0．01）。糖尿病组大鼠皮肤组织氧化应激指标含量均高于正常对照组，其中SOD和MPO差异有统计学意义（t值分别为4．4、3．7，P值均小于0．05）；氨基胍治疗组各氧化应激指标含量均较糖尿病组降低，其中SOD含量显著低于糖尿病组（t=-1．4，P〈0．05）；基质治疗组MDA、MPO含量显著低于糖尿病组（t值分别为2．6、2．9，P值均小于0．05）。结论外用氨基胍霜剂可以在一定程度上阻碍糖尿病大鼠皮肤组织中AGE的形成，改善表皮KC细胞增殖能力，适当降低皮肤组织氧化应激状态；单用霜剂基质也可适当降低皮肤组织氧化应激状态。
Objective To investigate the effects of aminoguanidine cream on the proliferation of keratinoeytes （KC） , content of advanced glycosylation end products （AGE） and oxidative stress in skin tissue of rats with diabetes. Methods Stearie acid, liquid paraffin, vaseline, lanolin, isopropyl myristate fat, glycerol, 50 g/L alcohol paraben, aminoguanidine hydrochloride etc. were mixed in certain proportion to make aminoguanidine eream, and cream without aminoguanidine was used as matrix. The dorsal skin of norreal rats were harvested and treated by aminoguanidine cream with dose of 5, 10 g/L, or 5 g/L together with 10 g/L azone. The transdermal effect was respectively measured at post treatment hour ：2, 4, 7, 10, 12, 24. Thirty SD rats were divided into normal control （ NC, n = 6） , diabetes （ D, n = 8 ） , aminoguanidine cream-interfered （AI, n = 8） , matrix cream-interfered groups （MI, n = 8） according to the random number table. Diabetes was reproduced by intraperitoneal injection of STZ （65 mg/kg） in rats of D, AI, and MI groups, and rats in NC group were injected with 0.05 mmol/L citrate buffer as control. One week later, dorsal skin of rats in AI and MI groups were respectively treated with 10 g/L aminoguanidine cream and matrix cream by external use for 4 weeks. AGE content was determined with fluorescence detection from skin colla- gen extract. KC cell cycle was detected by flow cytometry. Skin tissue specimens were obtained for determination of levels of superoxide dismutase （SOD）, malondialdehyde （MDA）, myeloperoxidase （MPO）, and total antioxidant capacity. Data were processed with t test. Results Transdermal effect of aminoguanidine cream with dose of 10 g/L was better than that with 5 g/L or 5 g/L ＋ 10 g/L azone cream. One rat was not induced successfully in MI group. Four weeks after model reproduction, 4 rats died in D group and 1 rat died in AI group. The AGE content in D group was obviously higher than that in NC group [ （36.8 ± 2.6） , （24.6 ±2.7） U per milligram hydroxyproline, respectively, t =7.2, P 〈0.01 ], and that in AI group [ （28.6 ± 3.7） U per milligram hydroxyproline ] was also lower as compared with that in D group （ t = - 3.9, P 〈 0.05 ）. There was no significant difference in AGE content between MI [ （ 32.2 ± 5.2） U per milligram hydroxyproline] and D groups （ t =1.6, P 〉0.05 ）. The percentage of KC in S phase was obviously lower in D group than in NC group [（5.3±0.6）%, （7.6±0.9）%, respectively, t =4.50, P 〈0.01], while that in MI group [ （9. 2 ± 1.5）% ] was higher as compared with that in D group （ t =4.90, P 〈0.01）. It was more higher in AI group than in D group on KC percentage in S and G2/M phase （ with t value respectively 6.80, 3.17, P values all below 0. 01 ）. The oxidative stress indexes of skin tissue in D group were all higher than those in NC group, in which levels of MPO and SOD showed statistical difference （with t value respectively 4.4, 3.7, P values all below 0.05 ）. The oxidative stress indexes were all lower in AI group than in D group, especially in SOD level （ t = -1.4, P 〈0.05）. Levels of MAD, MPOin MI group were significantly lower than those in D group （ with t value respectively 2.6, 2.9, P values all below 0. 05 ）. Conclusions Aminoguanidine cream can promote KC proliferation and appropriately reduce oxidative stress through inhibiting AGE formation to a certain extent in skin tissue of rats with diabetes. Signal use of matrix cream can also reduce oxidative stress in skin tissue of rats with diabetes.
出处
《中华烧伤杂志》
CAS
CSCD
北大核心
2011年第1期21-25,共5页
Chinese Journal of Burns
基金
基金项目：国家自然科学基金（30700871、30600645）
国家重点基础研究发展规划（2005CB522603）
关键词
糖尿病
糖基化终产物
高级
细胞增殖
氧化性应激
氨基胍
Diabetes mellitus
Glycosylation end products, advanced
Cell proliferation
Oxidative stress
Aminoguanidine
