负压创面治疗技术对糖尿病大鼠创面血管生成的影响
부압창면치료기술대당뇨병대서창면혈관생성적영향
Influence of negative pressure wound therapy on the angiogenesis of wounds in diabetic rats
  • 万方数据
    中华烧伤杂志 중화소상잡지
    16
    2013年 5期 442-447 ,共6页

    李小强%胡大海%刘洋%王耀军%韩夫%胡晓龙%李娜%张月%白晓智

    리소강%호대해%류양%왕요군%한부%호효룡%리나%장월%백효지

    负压伤口疗法%糖尿病%血管生成素类%血管形成 負壓傷口療法%糖尿病%血管生成素類%血管形成
    부압상구요법%당뇨병%혈관생성소류%혈관형성
    Negative-pressure wound therapy%Diabetes mellitus%Angiopoietins%Angiogenesis
    目的 观察负压创面治疗技术对糖尿病大鼠创面血管生成的影响. 方法 取40只SD大鼠,按65 mg/kg腹腔注射20 g/L链脲佐菌素诱导糖尿病模型.2周后按随机数字表法将大鼠分为对照组和负压组,每组20只,在大鼠背部正中切除2 cm×2 cm全层皮肤制备创面.伤后即刻,对照组创面常规换药；负压组创面予以每天4h持续负压(-16.0 kPa)治疗,连续7d.(1)治疗前及治疗后1、2周,分别采用血糖仪及电子秤检测2组大鼠血糖及体质量.(2)治疗前及治疗后1、3、7d,每组取5只大鼠,采用激光多普勒血流成像仪检测创面血流量.(3)治疗后3、7d,每组取5只大鼠处死后切取创面组织并分成两部分,取左侧组织行免疫组织化学染色观察血管形成情况,计算微血管密度.(4)取治疗前制备创面时切取的全层皮肤及治疗后3、7d冻存的右侧组织,治疗后1、14 d同前切取创面组织,采用实时荧光定量PCR法检测组织中血管内皮生长因子(VEGF)、血管内皮生长因子受体1(Fit-1)、血管生成素1(Ang-1)、Ang-2以及酪氨酸激酶受体2(Tie-2)mRNA的表达.对数据行双因素方差分析或LSD-t检验. 结果 (1)2组大鼠血糖及体质量水平总体或各时相点比较无明显差异(F值分别为0.667、0.176,t值为0.311 ～0.707,P值均大于0.05).(2)2组大鼠创面血流量总体比较有明显差异(F =24.66,P＜0.05).治疗后1、3、7d,负压组创面血流量分别为(179±24)、(219±12)、(192±30)灌注单位,显著高于对照组的(127±16)、(179±8)、(144±17)灌注单位(t值分别为3.71、5.57、2.77,P＜0.05或P＜0.01).(3)2组大鼠创面微血管密度总体比较有明显差异(F =33.25,P＜0.05).治疗后3d,负压组创面每100倍视野下微血管密度为(80±12)个,明显高于对照组的(38±4)个(t=9.257,P＜0.05).治疗后7d,2组大鼠创面微血管密度相近(t=1.159,P＞0.05),此时负压组血管排列规律、管腔宽畅,而对照组血管排列紊乱、管腔狭窄.(4)治疗后1、3d,负压组VEGF、Fit-1及Ang-1 mRNA表达水平均明显高于对照组(t值为1.28 ～11.60,P值均小于0.01)；治疗后7d,负压组Ang-1 mRNA表达水平(27.59±3.55)明显高于对照组(19.87±1.86,t=7.23,P＜0.001),其拮抗剂Ang-2 mRNA表达水平(5.79±0.61)明显低于对照组(17.62±0.85,t=19.88,P＜0.001).治疗后3～14 d,负压组Tie-2 mRNA表达水平均低于对照组(t值为8.92～15.60,P值均小于0.01). 结论 负压创面治疗技术可能通过增强创面愈合后期Ang-1表达以及降低Ang-2表达,促进糖尿病大鼠创面血管形成.
    목적 관찰부압창면치료기술대당뇨병대서창면혈관생성적영향. 방법 취40지SD대서,안65 mg/kg복강주사20 g/L련뇨좌균소유도당뇨병모형.2주후안수궤수자표법장대서분위대조조화부압조,매조20지,재대서배부정중절제2 cm×2 cm전층피부제비창면.상후즉각,대조조창면상규환약；부압조창면여이매천4h지속부압(-16.0 kPa)치료,련속7d.(1)치료전급치료후1、2주,분별채용혈당의급전자칭검측2조대서혈당급체질량.(2)치료전급치료후1、3、7d,매조취5지대서,채용격광다보륵혈류성상의검측창면혈류량.(3)치료후3、7d,매조취5지대서처사후절취창면조직병분성량부분,취좌측조직행면역조직화학염색관찰혈관형성정황,계산미혈관밀도.(4)취치료전제비창면시절취적전층피부급치료후3、7d동존적우측조직,치료후1、14 d동전절취창면조직,채용실시형광정량PCR법검측조직중혈관내피생장인자(VEGF)、혈관내피생장인자수체1(Fit-1)、혈관생성소1(Ang-1)、Ang-2이급락안산격매수체2(Tie-2)mRNA적표체.대수거행쌍인소방차분석혹LSD-t검험. 결과 (1)2조대서혈당급체질량수평총체혹각시상점비교무명현차이(F치분별위0.667、0.176,t치위0.311 ～0.707,P치균대우0.05).(2)2조대서창면혈류량총체비교유명현차이(F =24.66,P＜0.05).치료후1、3、7d,부압조창면혈류량분별위(179±24)、(219±12)、(192±30)관주단위,현저고우대조조적(127±16)、(179±8)、(144±17)관주단위(t치분별위3.71、5.57、2.77,P＜0.05혹P＜0.01).(3)2조대서창면미혈관밀도총체비교유명현차이(F =33.25,P＜0.05).치료후3d,부압조창면매100배시야하미혈관밀도위(80±12)개,명현고우대조조적(38±4)개(t=9.257,P＜0.05).치료후7d,2조대서창면미혈관밀도상근(t=1.159,P＞0.05),차시부압조혈관배렬규률、관강관창,이대조조혈관배렬문란、관강협착.(4)치료후1、3d,부압조VEGF、Fit-1급Ang-1 mRNA표체수평균명현고우대조조(t치위1.28 ～11.60,P치균소우0.01)；치료후7d,부압조Ang-1 mRNA표체수평(27.59±3.55)명현고우대조조(19.87±1.86,t=7.23,P＜0.001),기길항제Ang-2 mRNA표체수평(5.79±0.61)명현저우대조조(17.62±0.85,t=19.88,P＜0.001).치료후3～14 d,부압조Tie-2 mRNA표체수평균저우대조조(t치위8.92～15.60,P치균소우0.01). 결론 부압창면치료기술가능통과증강창면유합후기Ang-1표체이급강저Ang-2표체,촉진당뇨병대서창면혈관형성.
    Objective To observe the influence of negative pressure wound therapy on the angiogenesis of wounds in diabetic rats.Methods Diabetes model was reproduced by intraperitoneal injection of 20 g/L streptozotocin in the dosage of 65 mg/kg in 40 SD rats.Two weeks later,rats were divided into control group(C)and negative pressure group(NP)according to the random number table,with 20 rats in each group.A piece of full-thickness skin in the center of the back of each rat in the size of 2 cm × 2 cm was excised to produce a wound.Immediately after injury,wounds in group C were given conventional dressing change; wounds in group NP were treated with continuous negative pressure(-16.0 kPa)therapy for four hours a day,which lasted for seven days.(1)Blood glucose and body weight of rats in two groups were respectively measured by glucose meter and electronic scale before treatment,and 1 and 2 week(s)after.(2)Wound blood flow was detected by laser Doppler perfusion imager before treatment and on post treatment day(PTD)1,3,7,with 5 rats at each time point.(3)On PTD 3 and 7,respectively,five rats from each group were sacrificed.The wound tissue was excised and divided into two parts.The angiogenesis in the left part tissue was observed with immunohistochemical staining.The microvessel density was calculated.(4)The full-thickness skin excised before treatment and the right part tissue freezed on PTD 3 and 7 were collected.On PTD 1 and 14,wound tissue was excised in the above-mentioned method.The mRNA levels of the vascular endothelial growth factor(VEGF),vascular endothelial growth factor receptor 1(Fit-1),angiopoietin 1(Ang-1),Ang-2,and tyrosine kinase receptor 2(Tie-2)were determined with real-time fluorescence quantification PCR.Data were processed with two-way analysis of variance or LSD-t test.Results (1)No significant difference was observed between two groups in blood glucose level and body weight as a whole or at each time point(with F values respectively 0.667,0.176,t values from 0.311 to 0.707,P values all above 0.05).(2)The difference in the overall wound blood flow of rats between two groups was significant(F =24.66,P ＜0.05).On PTD 1,3,7,values of wound blood flow of rats in group NPwere (179 ±24),(219 ± 12),(192 ± 30)perfusion unit,significantly higher than those of rats in group C [(127 ±16),(179±8),(144±17)perfusion unit,with t values respectively 3.71,5.57,2.77,P ＜ 0.05 orP ＜0.01].(3)The difference in the overall microvessel density in the wound of rats between two groups was significant(F =33.25,P ＜ 0.05).On PTD 3,the microvessel density in the wound of rats in group NP was(80 ± 12)per 100-time visual field,which was significantly higher than that of group C [(38 ±4)per 100-time visual field,t =9.257,P ＜0.05].On PTD 7,the microvessel density in the wound of rats in two groups were close(t =1.159,P ＞0.05),but the vessels in group NP were regularly arranged with spacious lumen,while the vessels in group C were disorderly arranged with narrow lumen.(4)On PTD 1,3,mRNA expression levels of VEGF,Fit-1,and Ang-1 in group NP were obviously higher than those in group C(withtvalues from 1.28 to 11.60,Pvalues all below0.01).On PTD 7,the mRNA expression level of Ang-1(27.59 ± 3.55)in group NP was obviously higher than that in group C(19.87 ±1.86,t =7.23,P ＜ 0.001),while the mRNA level of its antagonist Ang-2(5.79 ± 0.61)in group NP was obviously lower than that in group C(17.62±0.85,t =19.88,P ＜0.001).On PTD 3,7,14,mRNA levels of Tie-2 in group NP were obviously lower than those in group C(with t values from 8.92 to 15.60,P values all below 0.01).Conclusions Negative pressure wound therapy may promote wound angiogenesis by enhancing the expression of Ang-1 and lowering the expression of Ang-2 in diabetic rats.
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