中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2014年
11期
785-789
,共5页
杨圣菊%孟国梁%顾黎雄%王建力
楊聖菊%孟國樑%顧黎雄%王建力
양골국%맹국량%고려웅%왕건력
皮肤%糖尿病%胶原Ⅰ型%胶原Ⅲ型%糖基化终产物,高级
皮膚%糖尿病%膠原Ⅰ型%膠原Ⅲ型%糖基化終產物,高級
피부%당뇨병%효원Ⅰ형%효원Ⅲ형%당기화종산물,고급
Skin%Diabetes mellitus%Collagen type Ⅰ%Collagen typeⅢ%Glycosylation end products,advanced
目的 观察不同病程糖尿病小鼠皮肤组织晚期糖基化终末产物(AGE)的表达及其对胶原纤维的影响.方法 健康8周龄C57BL/6J雄性小鼠采用小剂量链佐星(50 mg/kg)多次注射法建立糖尿病模型,分别在造模后第4周和12周处死小鼠,取背部中央皮肤,HE染色观察组织学结构,样本碱水解法测定羟脯氨酸水平反映胶原总量,限制性胃蛋白酶降解法测定胶原交联程度,实时定量PCR法测定Ⅰ型胶原和Ⅲ型胶原基因表达,荧光分光光度法和Western印迹检测晚期糖基化终末产物水平,硫代巴比妥酸法测定丙二醛含量.统计学分析采用t检验.结果 镜下可见糖尿病小鼠皮肤组织明显变薄,胶原呈进行性减少.第4周和12周时,糖尿病组小鼠皮肤组织羟脯氨酸含量与正常对照组比较显著降低[(684.5±76.7)比(787.7±87.7) mg/g,(558.1±73.1)比(757.8±75.3) mg/g,均P＜ 0.01],皮肤AGE水平显著上升[(37.47±10.65)比(26.39±3.74)AUF/mg羟脯氨酸,(47.70±5.66)比(29.91±6.50) AUF/mg羟脯氨酸,均P＜ 0.01],丙二醛含量显著增加[(6.62±0.47)比(4.82±0.56) μmol/L,(8.63±0.36)比(5.15±0.46)μmol/L,均P＜ 0.01],交联胶原以及Ⅰ、Ⅲ型胶原mRNA表达均下降(P＜0.01或0.05),糖尿病12周组非交联胶原亦明显下降(P＜0.05);与糖尿病4周组相比,糖尿病12周组小鼠皮肤羟脯氨酸含量、Ⅰ型胶原表达进一步减少(P＜0.05),AGE和丙二醛水平进一步增加(P＜0.01).结论 糖尿病小鼠皮肤组织胶原纤维性状发生改变,可能与皮肤组织中晚期糖基化终末产物水平上升及氧化损伤加剧有关.
目的 觀察不同病程糖尿病小鼠皮膚組織晚期糖基化終末產物(AGE)的錶達及其對膠原纖維的影響.方法 健康8週齡C57BL/6J雄性小鼠採用小劑量鏈佐星(50 mg/kg)多次註射法建立糖尿病模型,分彆在造模後第4週和12週處死小鼠,取揹部中央皮膚,HE染色觀察組織學結構,樣本堿水解法測定羥脯氨痠水平反映膠原總量,限製性胃蛋白酶降解法測定膠原交聯程度,實時定量PCR法測定Ⅰ型膠原和Ⅲ型膠原基因錶達,熒光分光光度法和Western印跡檢測晚期糖基化終末產物水平,硫代巴比妥痠法測定丙二醛含量.統計學分析採用t檢驗.結果 鏡下可見糖尿病小鼠皮膚組織明顯變薄,膠原呈進行性減少.第4週和12週時,糖尿病組小鼠皮膚組織羥脯氨痠含量與正常對照組比較顯著降低[(684.5±76.7)比(787.7±87.7) mg/g,(558.1±73.1)比(757.8±75.3) mg/g,均P＜ 0.01],皮膚AGE水平顯著上升[(37.47±10.65)比(26.39±3.74)AUF/mg羥脯氨痠,(47.70±5.66)比(29.91±6.50) AUF/mg羥脯氨痠,均P＜ 0.01],丙二醛含量顯著增加[(6.62±0.47)比(4.82±0.56) μmol/L,(8.63±0.36)比(5.15±0.46)μmol/L,均P＜ 0.01],交聯膠原以及Ⅰ、Ⅲ型膠原mRNA錶達均下降(P＜0.01或0.05),糖尿病12週組非交聯膠原亦明顯下降(P＜0.05);與糖尿病4週組相比,糖尿病12週組小鼠皮膚羥脯氨痠含量、Ⅰ型膠原錶達進一步減少(P＜0.05),AGE和丙二醛水平進一步增加(P＜0.01).結論 糖尿病小鼠皮膚組織膠原纖維性狀髮生改變,可能與皮膚組織中晚期糖基化終末產物水平上升及氧化損傷加劇有關.
목적 관찰불동병정당뇨병소서피부조직만기당기화종말산물(AGE)적표체급기대효원섬유적영향.방법 건강8주령C57BL/6J웅성소서채용소제량련좌성(50 mg/kg)다차주사법건립당뇨병모형,분별재조모후제4주화12주처사소서,취배부중앙피부,HE염색관찰조직학결구,양본감수해법측정간포안산수평반영효원총량,한제성위단백매강해법측정효원교련정도,실시정량PCR법측정Ⅰ형효원화Ⅲ형효원기인표체,형광분광광도법화Western인적검측만기당기화종말산물수평,류대파비타산법측정병이철함량.통계학분석채용t검험.결과 경하가견당뇨병소서피부조직명현변박,효원정진행성감소.제4주화12주시,당뇨병조소서피부조직간포안산함량여정상대조조비교현저강저[(684.5±76.7)비(787.7±87.7) mg/g,(558.1±73.1)비(757.8±75.3) mg/g,균P＜ 0.01],피부AGE수평현저상승[(37.47±10.65)비(26.39±3.74)AUF/mg간포안산,(47.70±5.66)비(29.91±6.50) AUF/mg간포안산,균P＜ 0.01],병이철함량현저증가[(6.62±0.47)비(4.82±0.56) μmol/L,(8.63±0.36)비(5.15±0.46)μmol/L,균P＜ 0.01],교련효원이급Ⅰ、Ⅲ형효원mRNA표체균하강(P＜0.01혹0.05),당뇨병12주조비교련효원역명현하강(P＜0.05);여당뇨병4주조상비,당뇨병12주조소서피부간포안산함량、Ⅰ형효원표체진일보감소(P＜0.05),AGE화병이철수평진일보증가(P＜0.01).결론 당뇨병소서피부조직효원섬유성상발생개변,가능여피부조직중만기당기화종말산물수평상승급양화손상가극유관.
Objective To investigate the expressions of advanced glycosylation end products (AGE) in skin of mice with diabetes mellitus (DM) for different durations,and to evaluate their influence on collagen fibers.Methods Forty healthy 8-week-old male C57BL/6J mice were divided into DM group (n =20) and control group (n =20) to receive multiple intraperitoneal injections of low dose streptozotocin (50 mg/kg) and citric acid buffer (0.1 mol/L),respectively,for 5 consecutive days.Ten mice were sacrificed in each group on week 4 and 12 respectively after the last intraperitoneal injection,and full-thickness skin tissue samples were harvested from the middorsal region of each mouse.Then,hematoxylin-eosin (HE) staining was performed to observe histological changes,and total collagen content was estimated according to hydroxyproline content measured by an alkalinehydrolysis method.The cross-linking degree of collagen was determined by Edman degradation method using pepsin,the mRNA expression level of collagen type Ⅰ and Ⅲ by real-time quantitative PCR,the content of AGE by fluorospectrophotometry and Western blotting,and the level of malondialdehyde (MDA) by using a thiobarbituric acid method.Statistical analysis was carried out by t test.Results As light microscopy showed,the skin became obviously thinner in the diabetic mice with a progressive decrease in the number of collagen fibers in comparison with the control mice.On week 4 and 12 after the last injection,the diabetic mice exhibited a significant reduction in the content of hydroxyproline ((684.5 ± 76.7) vs.(787.7 ± 87.7) rg/g,(558.1 ± 73.1) vs.(757.8 ± 75.3) mg/g,both P ＜ 0.01) and in the levels of cross-linked collagen as well as mRNA expressions of collagen Ⅰ and Ⅲ (P ＜ 0.01 or 0.05),but a significant increase in the content of AGE ((37.47 ± 10.65) vs.(26.39 ± 3.74) AUF/mg hydroxyproline,(47.70 ± 5.66) vs.(29.91 ± 6.50) AUF/mg hydroxyproline,both P ＜ 0.01) and MDA ((6.62 ± 0.47) vs.(4.82 ± 0.56) μmol/L,(8.63 ± 0.36) vs.(5.15 ± 0.46) μmol/L,both P＜ 0.01) in skin tissue,compared with the control mice.The level of non-cross-linked collagen in skin tissue was also lower in the diabetic mice than in the control mice on week 12 (P ＜ 0.05).Moreover,the contents of hydroxyproline and the expression levels of collagen I in skin were significantly lower (P ＜ 0.05),but the levels of AGE and MDA were significantly higher (P ＜ 0.01) in the diabetic mice on week 12 than in those on week 4.Conclusions The characteristics of collagen fibers in skin are altered in diabetic mice when compared with normal control mice,which may be associated with increased AGE content and oxidative injury in skin.
