中华烧伤杂志
中華燒傷雜誌
중화소상잡지
16
2012年
1期
32-35
,共4页
牛轶雯%缪明远%董炜%董叫云%曹晓赞%陆树良
牛軼雯%繆明遠%董煒%董叫雲%曹曉讚%陸樹良
우질문%무명원%동위%동규운%조효찬%륙수량
糖尿病%氧化性应激%糖基化终产物,高级%皮肤%创面愈合
糖尿病%氧化性應激%糖基化終產物,高級%皮膚%創麵愈閤
당뇨병%양화성응격%당기화종산물,고급%피부%창면유합
Diabetes Mellitus%Oxidative stress%Glycosylation end products,advanced%Skin%Wound healing
目的 观察糖尿病患者皮肤和创面中晚期糖基化终末产物(AGE)的蓄积和炎症反应,并结合体外干预实验推测两者之间可能存在的关系. 方法 采集10例2型糖尿病患者(糖尿病组)和12例非糖尿病患者(非糖对照组)的皮肤和创面组织标本,部分于光学显微镜下观察胶原排列、细胞分布(HE染色),AGE及其受体(RAGE)表达(免疫组织化学染色)；部分匀浆后采用ELISA法检测丙二醛含量.体外采集、分离健康人外周血中性粒细胞,并分为正常对照组(添加RPMI-1640培养液培养)、低浓度干预组、中浓度干预组和高浓度干预组,3个干预组分别在RPMI-1640培养液中添加AGE修饰的人血清白蛋白,其中AGE的浓度依次为0.315、0.625、1.250 mg/mL;噻唑蓝法检测细胞存活率,荧光探针二氢二氯荧光黄双乙酸钠测定细胞内活性氧水平.对数据进行t检验. 结果 与非糖对照组比较,糖尿病组皮肤组织中胶原萎缩,排列紊乱；创面组织中炎性细胞弥散分布,胶原排列稀疏紊乱.糖尿病组皮肤组织中AGE和RAGE表达均多于非糖对照组；2组创面组织中RAGE呈阳性表达的细胞数量均多于所在组皮肤组织,以糖尿病组更显著,其创面内可见大量RAGE呈阳性表达的炎性细胞.糖尿病组患者皮肤和创面组织中每毫克蛋白丙二醛含量分别为(6 3±1.0)、(7.1±2.4)nmol,均明显高于非糖对照组相应组织[(2.9±1.0)、(3.6±1.4)nmol,t值分别为8.017、4.349,P ＜0.05或P＜0.01].与正常对照组[(34±5)％]比较,低浓度干预组、中浓度干预组、高浓度干预组中性粒细胞存活率均明显上升[(59±8)％、(77±5)％、( 67±6)％,t值分别为7.195、14.890、11.130,P＜0.05或P＜0.01].与正常对照组(0.58±0.06)比较,低浓度干预组、中浓度干预组、高浓度干预组中性粒细胞内活性氧水平均升高(1.67±0.14、2.13±0.17、3 48±0.48,t值分别为20.195、24.905、16.864,P＜0.05或P＜0.01). 结论 异常的氧化应激水平导致糖尿病皮肤具有异常的创伤修复起点,创面愈合过程中AGE-RAGE效应是影响糖尿病创面氧化应激水平的重要因素.
目的 觀察糖尿病患者皮膚和創麵中晚期糖基化終末產物(AGE)的蓄積和炎癥反應,併結閤體外榦預實驗推測兩者之間可能存在的關繫. 方法 採集10例2型糖尿病患者(糖尿病組)和12例非糖尿病患者(非糖對照組)的皮膚和創麵組織標本,部分于光學顯微鏡下觀察膠原排列、細胞分佈(HE染色),AGE及其受體(RAGE)錶達(免疫組織化學染色)；部分勻漿後採用ELISA法檢測丙二醛含量.體外採集、分離健康人外週血中性粒細胞,併分為正常對照組(添加RPMI-1640培養液培養)、低濃度榦預組、中濃度榦預組和高濃度榦預組,3箇榦預組分彆在RPMI-1640培養液中添加AGE脩飾的人血清白蛋白,其中AGE的濃度依次為0.315、0.625、1.250 mg/mL;噻唑藍法檢測細胞存活率,熒光探針二氫二氯熒光黃雙乙痠鈉測定細胞內活性氧水平.對數據進行t檢驗. 結果 與非糖對照組比較,糖尿病組皮膚組織中膠原萎縮,排列紊亂；創麵組織中炎性細胞瀰散分佈,膠原排列稀疏紊亂.糖尿病組皮膚組織中AGE和RAGE錶達均多于非糖對照組；2組創麵組織中RAGE呈暘性錶達的細胞數量均多于所在組皮膚組織,以糖尿病組更顯著,其創麵內可見大量RAGE呈暘性錶達的炎性細胞.糖尿病組患者皮膚和創麵組織中每毫剋蛋白丙二醛含量分彆為(6 3±1.0)、(7.1±2.4)nmol,均明顯高于非糖對照組相應組織[(2.9±1.0)、(3.6±1.4)nmol,t值分彆為8.017、4.349,P ＜0.05或P＜0.01].與正常對照組[(34±5)％]比較,低濃度榦預組、中濃度榦預組、高濃度榦預組中性粒細胞存活率均明顯上升[(59±8)％、(77±5)％、( 67±6)％,t值分彆為7.195、14.890、11.130,P＜0.05或P＜0.01].與正常對照組(0.58±0.06)比較,低濃度榦預組、中濃度榦預組、高濃度榦預組中性粒細胞內活性氧水平均升高(1.67±0.14、2.13±0.17、3 48±0.48,t值分彆為20.195、24.905、16.864,P＜0.05或P＜0.01). 結論 異常的氧化應激水平導緻糖尿病皮膚具有異常的創傷脩複起點,創麵愈閤過程中AGE-RAGE效應是影響糖尿病創麵氧化應激水平的重要因素.
목적 관찰당뇨병환자피부화창면중만기당기화종말산물(AGE)적축적화염증반응,병결합체외간예실험추측량자지간가능존재적관계. 방법 채집10례2형당뇨병환자(당뇨병조)화12례비당뇨병환자(비당대조조)적피부화창면조직표본,부분우광학현미경하관찰효원배렬、세포분포(HE염색),AGE급기수체(RAGE)표체(면역조직화학염색)；부분균장후채용ELISA법검측병이철함량.체외채집、분리건강인외주혈중성립세포,병분위정상대조조(첨가RPMI-1640배양액배양)、저농도간예조、중농도간예조화고농도간예조,3개간예조분별재RPMI-1640배양액중첨가AGE수식적인혈청백단백,기중AGE적농도의차위0.315、0.625、1.250 mg/mL;새서람법검측세포존활솔,형광탐침이경이록형광황쌍을산납측정세포내활성양수평.대수거진행t검험. 결과 여비당대조조비교,당뇨병조피부조직중효원위축,배렬문란；창면조직중염성세포미산분포,효원배렬희소문란.당뇨병조피부조직중AGE화RAGE표체균다우비당대조조；2조창면조직중RAGE정양성표체적세포수량균다우소재조피부조직,이당뇨병조경현저,기창면내가견대량RAGE정양성표체적염성세포.당뇨병조환자피부화창면조직중매호극단백병이철함량분별위(6 3±1.0)、(7.1±2.4)nmol,균명현고우비당대조조상응조직[(2.9±1.0)、(3.6±1.4)nmol,t치분별위8.017、4.349,P ＜0.05혹P＜0.01].여정상대조조[(34±5)％]비교,저농도간예조、중농도간예조、고농도간예조중성립세포존활솔균명현상승[(59±8)％、(77±5)％、( 67±6)％,t치분별위7.195、14.890、11.130,P＜0.05혹P＜0.01].여정상대조조(0.58±0.06)비교,저농도간예조、중농도간예조、고농도간예조중성립세포내활성양수평균승고(1.67±0.14、2.13±0.17、3 48±0.48,t치분별위20.195、24.905、16.864,P＜0.05혹P＜0.01). 결론 이상적양화응격수평도치당뇨병피부구유이상적창상수복기점,창면유합과정중AGE-RAGE효응시영향당뇨병창면양화응격수평적중요인소.
Objective To investigate the accumulation of advanced glycation end products(AGE)and the inflammatory response of skin and wound in diabetic patients,and to analyze their relationship in vitro. Methods Histological staining and immunohistochemical staining was respectively performed on skin and wound tissue specimens collected from 10 patients with Type Ⅱ diabetes mellitus(diabetes group)and 12 non-diabetic patients with skin injury(control group)to observe the arrangement of collagen and the distribution of inflammatory cells,and to determine the expression levels of AGE and its receptor( RAGE).Malondialdehyde(MDA)levels in skin and wound tissue homogenates were assayed by enzyme-linked immunosorbent assay.In vitro,human neutrophils were isolated and treated with RPMI-1640 culture medium or that containing AGE-human serum albumin in the concentration of 0.315,0.625,1.250 mg/mL,and they were identified as normal control(NC)group,low concentration(L)group,moderate concentration (M)group,and high concentration(H)group.Cell viability in each group was determined by MTT colorimetric assay,and the reactive oxygen species(ROS)in cell was measured with 2′,7′-dichlorfluorescein-diacetate.Data were processed with t test. Results Compared with those of skin in control group,collagens of skin tissues in diabetes group atrophied and disorderly arranged.Inflammatory cells in wounds in diabetes group were dispersed,in which collagens arranged loosely and irregularly,as compared with those of wounds in control group.Expression levels of AGE and RAGE of skin in diabetes group were higher than those in control group.In diabetes and control groups,especially in diabetes group,the numbers of RAGE-positive cells in wound tissue were more than those in skin tissue.Large amount of inflammatory cells with positive expression of RAGE were observed in diabetes group.MDA level of skin and wound tissue in diabetes group was respectively(6.3 ± 1.0),(7.1 ± 2.4)nmol per milligram protein,which were obviously higher than those in control group [(2.9 ± 1.0),( 3.6 ± 1.4)nmol per milligram protein,with t value respectively 8.017,4.349,P ＜0.05 or P ＜ 0.01 ].Cell viability and ROS levels in neutrophils were increased in L,M,and H groups [(59 ±8)％,(77 ±5)％,(67 ±6)％ and 1.67 ±0.14,2.13 ±0.17,3.48 ±0.48] as compared with those in NC group [(34 ± 5)％ and 0.58 ± 0.06,with t value respectively 7.195,14.890,11.130 and 20.195,24.905,16.864,P ＜ 0.05 or P ＜ 0.01 ]. Conclusions Abnormal oxidative stress in diabetic skin leads to an atypical origin of wound repair.AGE-RAGE effect is a critical mediator for oxidative stress in diabetic wound tissue during wound healing.
