CAJ | 학술논문

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、50 g/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、10 g/L氨基胍霜剂和5 g/L氨基胍+10g/L氮酮霜剂处理,于用药后2、4、7、10、12、24 h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65 mg/kg,诱导糖尿病模型;对照组大鼠注射0.05 mmol/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10 g/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 10 g/L氨基胍霜剂透皮效果优于5g/L氨基胍和5 g/L氨基胍+10 g/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. 目的觀察外敷氨基胍霜劑對糖尿病大鼠皮膚組織晚期糖基化終末產物(AGE)形成、KC細胞增殖及氧化應激的影響.方法將硬脂痠、液狀石蠟、凡士林、羊毛脂、肉豆蔻痠異丙酯、甘油、50 g/L尼泊金醇、鹽痠氨基胍等試劑按一定比例混閤製成氨基胍霜劑,以不含有氨基胍的霜劑為基質.取健康大鼠揹部皮膚,分彆用5、10 g/L氨基胍霜劑和5 g/L氨基胍+10g/L氮酮霜劑處理,于用藥後2、4、7、10、12、24 h測定藥物透皮效果.將30隻SD大鼠按隨機數字錶法分為正常對照組6隻、糖尿病組8隻、氨基胍治療組8隻、基質治療組8隻.後3組大鼠腹腔註射鏈脲佐菌素65 mg/kg,誘導糖尿病模型;對照組大鼠註射0.05 mmol/L檸檬痠緩遲液.註射1週後,正常對照組與糖尿病組大鼠不行任何治療,氨基胍治療組與基質治療組大鼠揹部分彆連續外用10 g/L氨基胍霜劑與基質治療4週.取各組皮膚組織,膠原提取液熒光彊度檢測法測定AGE含量,流式細胞儀分析錶皮KC週期,檢測氧化應激相關指標超氧化物歧化酶(SOD)、丙二醛、總抗氧化能力、髓過氧化物酶(MPO)含量.對實驗數據行t檢驗.結果 10 g/L氨基胍霜劑透皮效果優于5g/L氨基胍和5 g/L氨基胍+10 g/L氮酮的霜劑.1隻基質治療組大鼠未誘導成功.建模後4週,糖尿病組與氨基胍治療組大鼠分彆死亡4隻和1隻.糖尿病組大鼠皮膚組織AGE含量為每毫剋羥脯氨痠(OHP)中(36.8±2.6)U,明顯高于正常對照組的每毫剋OHP中(24.6±2.7)U(t=7.2,P＜0.01);氨基胍治療組AGE含量為每毫剋OHP中(28.6±3.7)U,明顯低于糖尿病組(t=-3.9,P＜0.05);基質治療組AGE含量[每毫剋OHP中(32.2±5.2)U]與糖尿病組相近(t=1.6,P＞0.05).糖尿病組大鼠S期KC比例為(5.3±0.6)%,低于正常對照組的(7.6±0.9)%(t=4.50,P＜0.01);氨基胍治療組大鼠S期和G2/M期KC比例均明顯高于糖尿病組(t值分彆為6.80、3.17,P值均小于0.01);基質治療組大鼠S期KC比例[(9.2±1.5)%]顯著高于糖尿病組(t=4.90,P＜0.01).糖尿病組大鼠皮膚組織氧化應激指標含量均高于正常對照組,其中SOD和MPO差異有統計學意義(t值分彆為4.4、3.7,P值均小于0.05);氨基胍治療組各氧化應激指標含量均較糖尿病組降低,其中SOD含量顯著低于糖尿病組(t=-1.4,P＜0.05);基質治療組MDA、MPO含量顯著低于糖尿病組(t值分彆為2.6、2.9,P值均小于0.05).結論外用氨基胍霜劑可以在一定程度上阻礙糖尿病大鼠皮膚組織中AGE的形成,改善錶皮KC細胞增殖能力,適噹降低皮膚組織氧化應激狀態;單用霜劑基質也可適噹降低皮膚組織氧化應激狀態.
목적 관찰외부안기고상제대당뇨병대서피부조직만기당기화종말산물(AGE)형성、KC세포증식급양화응격적영향.방법 장경지산、액상석사、범사림、양모지、육두구산이병지、감유、50 g/L니박금순、염산안기고등시제안일정비례혼합제성안기고상제,이불함유안기고적상제위기질.취건강대서배부피부,분별용5、10 g/L안기고상제화5 g/L안기고+10g/L담동상제처리,우용약후2、4、7、10、12、24 h측정약물투피효과.장30지SD대서안수궤수자표법분위정상대조조6지、당뇨병조8지、안기고치료조8지、기질치료조8지.후3조대서복강주사련뇨좌균소65 mg/kg,유도당뇨병모형;대조조대서주사0.05 mmol/L저몽산완충액.주사1주후,정상대조조여당뇨병조대서불행임하치료,안기고치료조여기질치료조대서배부분별련속외용10 g/L안기고상제여기질치료4주.취각조피부조직,효원제취액형광강도검측법측정AGE함량,류식세포의분석표피KC주기,검측양화응격상관지표초양화물기화매(SOD)、병이철、총항양화능력、수과양화물매(MPO)함량.대실험수거행t검험.결과 10 g/L안기고상제투피효과우우5g/L안기고화5 g/L안기고+10 g/L담동적상제.1지기질치료조대서미유도성공.건모후4주,당뇨병조여안기고치료조대서분별사망4지화1지.당뇨병조대서피부조직AGE함량위매호극간포안산(OHP)중(36.8±2.6)U,명현고우정상대조조적매호극OHP중(24.6±2.7)U(t=7.2,P＜0.01);안기고치료조AGE함량위매호극OHP중(28.6±3.7)U,명현저우당뇨병조(t=-3.9,P＜0.05);기질치료조AGE함량[매호극OHP중(32.2±5.2)U]여당뇨병조상근(t=1.6,P＞0.05).당뇨병조대서S기KC비례위(5.3±0.6)%,저우정상대조조적(7.6±0.9)%(t=4.50,P＜0.01);안기고치료조대서S기화G2/M기KC비례균명현고우당뇨병조(t치분별위6.80、3.17,P치균소우0.01);기질치료조대서S기KC비례[(9.2±1.5)%]현저고우당뇨병조(t=4.90,P＜0.01).당뇨병조대서피부조직양화응격지표함량균고우정상대조조,기중SOD화MPO차이유통계학의의(t치분별위4.4、3.7,P치균소우0.05);안기고치료조각양화응격지표함량균교당뇨병조강저,기중SOD함량현저저우당뇨병조(t=-1.4,P＜0.05);기질치료조MDA、MPO함량현저저우당뇨병조(t치분별위2.6、2.9,P치균소우0.05).결론 외용안기고상제가이재일정정도상조애당뇨병대서피부조직중AGE적형성,개선표피KC세포증식능력,괄당강저피부조직양화응격상태;단용상제기질야가괄당강저피부조직양화응격상태.
Objective To investigate the effects of aminoguanidine cream on the proliferation of keratinocytes (KC), content of advanced glycosylation end products (AGE) and oxidative stress in skin tissue of rats with diabetes. Methods Stearic 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 cream, and cream without aminoguanidine was used as matrix. The dorsal skin of normal 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 collagen 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, MPO in 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.