负压伤口疗法对糖尿病足不同程度缺血创面肉芽组织Ⅰ型胶原蛋白表达的影响

doi:10.3877/cma.j.issn.1673-9450.2018.01.006

中华损伤与修复杂志(电子版) ›› 2018, Vol. 13 ›› Issue (01) : 24 -29. doi: 10.3877/cma.j.issn.1673-9450.2018.01.006

所属专题：文献；

论著

负压伤口疗法对糖尿病足不同程度缺血创面肉芽组织Ⅰ型胶原蛋白表达的影响

杨少玲¹^,^†(

), 朱旅云¹, 胡丽叶¹, 刘洋², 郭炳彦³, 窦京涛⁴

^1. 050082　石家庄，解放军白求恩国际和平医院内分泌科
^2. 050017　石家庄，河北医科大学研究生学院
^3. 050017　石家庄，河北医科大学第二医院心血管内科
^4. 100853　北京，解放军总医院内分泌科

收稿日期:2017-12-25 出版日期:2018-02-01
通信作者: 杨少玲
基金资助:
河北省科技支撑计划项目(12277725); 河北省自然科学基金项目(H2013505067)

Effects of negative pressure wound therapy on the expression of typeⅠcollagen in granulation tissue of different ischemic wound of diabetic foot

Shaoling Yang¹^,^†(), Lyuyun Zhu¹, Liye Hu¹, Yang Liu², Bingyan Guo³, Jingtao Dou⁴

^1. Department of Endocrinology, Bethune International Peace Hospital of People′s Liberation Army, Shijiazhuang 050082, China
^2. Graduate School of Hebei Medical University, Shijiazhuang 050017, China
^3. Department of Cardiovascular Medicine, Second Hospital of Hebei Medical University, Shijiazhuang 050017, China
^4. Department of Endocrinology, People′s Liberation Army General Hospital, Beijing 100853, China

Received:2017-12-25 Published:2018-02-01
Corresponding author: Shaoling Yang
About author:
Corresponding author: Yang Shaoling, Email: yshling9@126.com

全文 ( ) 下载PDF ( ) 导出引用

引用本文：

杨少玲, 朱旅云, 胡丽叶, 刘洋, 郭炳彦, 窦京涛. 负压伤口疗法对糖尿病足不同程度缺血创面肉芽组织Ⅰ型胶原蛋白表达的影响[J/OL]. 中华损伤与修复杂志(电子版), 2018, 13(01): 24-29.

Shaoling Yang, Lyuyun Zhu, Liye Hu, Yang Liu, Bingyan Guo, Jingtao Dou. Effects of negative pressure wound therapy on the expression of typeⅠcollagen in granulation tissue of different ischemic wound of diabetic foot[J/OL]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2018, 13(01): 24-29.

目的

探讨负压伤口疗法(NPWT)对糖尿病足不同程度缺血创面肉芽组织Ⅰ型胶原蛋白表达的影响。

方法

选取2014年1月至2016年1月在解放军白求恩国际和平医院住院期间符合入组标准的糖尿病足患者30例，依据踝肱指数(ABI)把入组患者分为轻度下肢缺血组(0.7≤ABI≤0.9)和中度下肢缺血组(0.5≤ABI<0.7)，每组各15例。两组患者均给予NPWT治疗，疗程14 d。分别于治疗前及治疗第14天取患者创面肉芽组织，将标本分成2份，1份固定在4%多聚甲醛中用于Ⅰ型胶原蛋白免疫组织化学染色，观察Ⅰ型胶原蛋白沉积情况；另1份于－80 ℃保存，用Western blotting法和Real-time PCR法定量测定Ⅰ型胶原蛋白及其mRNA表达情况。数据比较采用独立样本t检验。

结果

免疫组织化学染色分析显示，治疗第14天，两组Ⅰ型胶原蛋白平均面密度值较治疗前均明显升高，差异均有统计学意义(P值均小于0.05)；但与中度下肢缺血组(0.25±0.06)相比，轻度下肢缺血组胶原蛋白平均面密度值(0.35±0.07)升高更为明显，差异有统计学意义(t＝4.71，P<0.05)。Western blotting法分析显示，治疗第14天，两组Ⅰ型胶原蛋白相对表达水平较治疗前均明显升高，差异均有统计学意义(P值均小于0.05);但与中度下肢缺血组(0.16±0.09)相比，轻度下肢缺血组Ⅰ型胶原蛋白相对表达水平(0.28±0.08)升高更为明显，差异有统计学意义(t＝3.51，P<0.05)。Real-time PCR法分析显示，治疗第14天，两组Ⅰ型胶原蛋白mRNA相对表达水平较治疗前均明显升高，差异均有统计学意义(P值均小于0.05);与中度下肢缺血组(2.63±0.62)相比，轻度下肢缺血组Ⅰ型胶原蛋白mRNA相对表达水平(3.83±0.60)升高更为明显，差异有统计学意义(t＝5.33，P<0.05)。

结论

NPWT对糖尿病足患者不同程度缺血创面Ⅰ型胶原蛋白表达的影响不同，对于轻度缺血创面肉芽组织的Ⅰ型胶原蛋白的表达增加效果较好，随着缺血程度的加重，NPWT对创面肉芽组织的Ⅰ型胶原蛋白的表达促进效果可能会下降。

关键词: 负压伤口疗法, 糖尿病足, 缺血, 肉芽组织, Ⅰ型胶原蛋白

Objective

To investigate the effect of negative pressure wound therapy (NPWT) on the expression of type Ⅰcollagen in granulation tissue of different ischemic wound of diabetic foot.

Methods

From January 2014 to January 2016, 30 patients at Bethune International Peace Hospital of People′s Liberation Army with diabetic foot wounds fitting the inclusion criteria were found and enrolled in the clinical trial. The patients were divided into mild lower extremity ischemia group and moderate lower extremity ischemia group according to the ankle-brachial index (ABI), 0.7≤ABI≤0.9 were mild lower extremity ischemia group and 0.5≤ABI<0.7 were moderate lower extremity ischemia group, each group had 15 cases. Both groups received NPWT for 14 days. Granulated tissue of wounds were collected before treatment and 14 days after treatment in both groups′ patients. All specimens were subdivided into two parts. One part was preserved in 4% paraformaldehyde for immunocytochemical staining of typeⅠcollagen to observe the deposition of type Ⅰ collagen. The other part was stored at -80 ℃ for measuring the expression of type Ⅰ collagen and its gene with Western blotting and Real-time PCR analysis. The data were analyzed by independent sample t test.

Results

The immunohistochemical analysis revealed that the mean area density of typeⅠcollagen enhanced in both two groups [(0.25±0.06) vs (0.35±0.07)]after they received NPWT, the differences were statistically significant (with P values below 0.05). But compared with the moderate lower extremity ischemia group, the degree of the increase in mild lower extremity ischemia group was more obvious, the difference was statistically significant (t=4.71, P<0.05). The Western blotting analysis showed that the relative protein expression value of typeⅠcollagen increased in both groups [(0.16±0.09) vs (0.28±0.08)] after they received NPWT, the differences were statistically significant (with P values below 0.05). However, the relative protein expression value of typeⅠcollagen increased obviously in mild lower extremity ischemia group compared with moderate lower extremity ischemia group, the difference was statistically significant (t=3.51, P<0.05). The Real time PCR analysis demonstrated that although the relative mRNA levels of typeⅠcollagen increased in both groups [(2.63±0.62) vs (3.83±0.60)] after they received NPWT, the differences were statistically significant (with P values below 0.05), the increase of relative expression value of mRNA in mild lower extremity ischemia group was more obvious than that in moderate lower extremity ischemia group, the difference was statistically significant (t=5.33, P<0.05).

Conclusions

NPWT had different effects on type Ⅰ collagen in different ischemic wounds of patients with diabetic foot. For the type I collagen of granulation tissue in mild ischemic wound, the expression increased better. With the aggravation of lower extremity ischemia, the effect of NPWT on the expression of type Ⅰ collagen in granulation tissue of diabetic foot wound may decrease.

Key words: Negative pressure wound therapy, Diabetic foot, Ischemia, Granulation tissue, TypeⅠcollagen

表1 两组糖尿病足患者一般资料比较

组别	例数	年龄(岁，±s)	性别(例)		空腹血糖(mmol/L，±s)	糖化血红蛋白(%，±s)
组别	例数	年龄(岁，±s)	男	女	空腹血糖(mmol/L，±s)	糖化血红蛋白(%，±s)
中度下肢缺血组	15	59.8±6.0	9	6	10.34±3.62	9.95±1.68
轻度下肢缺血组	15	60.1±6.9	6	9	9.75±2.09	10.13±1.63
检验值	?	0.06	?	?	0.56	0.33
P值	?	0.95^a	0.46^b		0.58^a	0.75^a
组别	例数	溃疡面积(cm²，±s)	溃疡时间(d，±s)	血肌酐(μmol/L，±s)	谷丙转氨酶(U/L，±s)
中度下肢缺血组	15	14.90±8.03	56.80±18.40	61.20±20.34	17.53±7.30
轻度下肢缺血组	15	18.60±11.10	55.01±17.60	62.00±16.49	19.66±7.44
检验值	?	1.06	0.28	0.11	0.80
P值	?	0.30^a	0.78^a	0.91^a	0.43^a

表1 两组糖尿病足患者一般资料比较

组别	例数	年龄(岁，±s)	性别(例)		空腹血糖(mmol/L，±s)	糖化血红蛋白(%，±s)
组别	例数	年龄(岁，±s)	男	女	空腹血糖(mmol/L，±s)	糖化血红蛋白(%，±s)
中度下肢缺血组	15	59.8±6.0	9	6	10.34±3.62	9.95±1.68
轻度下肢缺血组	15	60.1±6.9	6	9	9.75±2.09	10.13±1.63
检验值	?	0.06	?	?	0.56	0.33
P值	?	0.95^a	0.46^b		0.58^a	0.75^a
组别	例数	溃疡面积(cm²，±s)	溃疡时间(d，±s)	血肌酐(μmol/L，±s)	谷丙转氨酶(U/L，±s)
中度下肢缺血组	15	14.90±8.03	56.80±18.40	61.20±20.34	17.53±7.30
轻度下肢缺血组	15	18.60±11.10	55.01±17.60	62.00±16.49	19.66±7.44
检验值	?	1.06	0.28	0.11	0.80
P值	?	0.30^a	0.78^a	0.91^a	0.43^a

图1 两组糖尿病足患者NPWT治疗前后创面肉芽组织Ⅰ型胶原蛋白免疫组织化学染色情况(×400)。A示中度下肢缺血组治疗前Ⅰ型胶原蛋白表达；B示治疗第14天，中度下肢缺血组Ⅰ型胶原蛋白表达明显增多；C示轻度下肢缺血组治疗前Ⅰ型胶原蛋白表达；D示治疗第14天，轻度下肢缺血组Ⅰ型胶原蛋白表达明显增多；箭头示Ⅰ型胶原蛋白免疫组织化学染色阳性区域

表2 两组糖尿病足患者NPWT治疗前后创面肉芽组织Ⅰ型胶原蛋白平均面密度值统计结果(±s)

组别	例数	治疗前	治疗第14天	治疗前后变化值	t₁值	P₁值
中度下肢缺血组	15	0.12±0.02	0.36±0.06	0.25±0.06	14.31	<0.05
轻度下肢缺血组	15	0.10±0.02	0.46±0.06	0.35±0.07	21.11	<0.05
t₂值	?	1.52	4.29	4.71	?	?
P₂值	?	0.14	<0.05	<0.05	?	?

表2 两组糖尿病足患者NPWT治疗前后创面肉芽组织Ⅰ型胶原蛋白平均面密度值统计结果(±s)

组别	例数	治疗前	治疗第14天	治疗前后变化值	t₁值	P₁值
中度下肢缺血组	15	0.12±0.02	0.36±0.06	0.25±0.06	14.31	<0.05
轻度下肢缺血组	15	0.10±0.02	0.46±0.06	0.35±0.07	21.11	<0.05
t₂值	?	1.52	4.29	4.71	?	?
P₂值	?	0.14	<0.05	<0.05	?	?

图2 两组糖尿病足患者NPWT治疗前后创面肉芽组织Ⅰ型胶原蛋白Western blotting分析结果；NPWT：负压伤口疗法

表3 两组糖尿病足患者NPWT治疗前后创面肉芽组织Ⅰ型胶原蛋白相对表达水平统计结果(±s)

组别	例数	治疗前	治疗第14天	治疗前后变化值	t₁值	P₁值
中度下肢缺血组	15	0.38±0.05	0.53±0.08	0.16±0.09	6.39	<0.05
轻度下肢缺血组	15	0.33±0.07	0.62± 0.08	0.28±0.08	10.64	<0.05
t₂值	?	1.61	2.92	3.51	?	?
P₂值	?	0.12	0.01	<0.05	?	?

表3 两组糖尿病足患者NPWT治疗前后创面肉芽组织Ⅰ型胶原蛋白相对表达水平统计结果(±s)

组别	例数	治疗前	治疗第14天	治疗前后变化值	t₁值	P₁值
中度下肢缺血组	15	0.38±0.05	0.53±0.08	0.16±0.09	6.39	<0.05
轻度下肢缺血组	15	0.33±0.07	0.62± 0.08	0.28±0.08	10.64	<0.05
t₂值	?	1.61	2.92	3.51	?	?
P₂值	?	0.12	0.01	<0.05	?	?

图3 两组糖尿病足患者NPWT治疗前后创面肉芽组织Ⅰ型胶原蛋白mRNA相对表达水平

表4 两组糖尿病足患者接受NPWT治疗前后创面肉芽组织Ⅰ型胶原蛋白mRNA相对表达水平统计结果(±s)

组别	例数	治疗前	治疗第14天	治疗前后变化值	t₁值	P₁值
中度下肢缺血组	15	1.09±0.15	3.73±0.60	2.63±0.62	16.46	<0.05
轻度下肢缺血组	15	1.15±0.18	4.99±0.66	3.83±0.60	21.63	<0.05
t₂值	?	0.95	5.43	5.33	?	?
P₂值	?	0.35	<0.05	<0.05	?	?

表4 两组糖尿病足患者接受NPWT治疗前后创面肉芽组织Ⅰ型胶原蛋白mRNA相对表达水平统计结果(±s)

组别	例数	治疗前	治疗第14天	治疗前后变化值	t₁值	P₁值
中度下肢缺血组	15	1.09±0.15	3.73±0.60	2.63±0.62	16.46	<0.05
轻度下肢缺血组	15	1.15±0.18	4.99±0.66	3.83±0.60	21.63	<0.05
t₂值	?	0.95	5.43	5.33	?	?
P₂值	?	0.35	<0.05	<0.05	?	?

[1]	Ahmad J. The diabetic foot[J]. Diabetes Metab Syndr, 2016, 10(1):48-60.
[2]	Brownrigg JR, Griffin M, Hughes CO, et al. Influence of foot ulceration on cause-specific mortality in patients with diabetes mellitus[J]. J Vasc Surg, 2014, 60(4):982-986.
[3]	许樟荣. 糖尿病足病的诊治与预防[J/CD]. 中华损伤与修复杂志(电子版), 2014, 9(2):118-122.
[4]	关小宏. 关于我国糖尿病足防治策略的探讨[J/CD]. 中华损伤与修复杂志，2016, 11(2):84-89.
[5]	Blume PA, Walters J, Payne W, et al. Comparison of negative pressure wound therapy using vacuum-assisted closure with advanced moist wound therapy in the treatment of diabetic foot ulcers: a multicenter randomized controlled trial[J]. Diabetes Care, 2008, 31(4):631-636.
[6]	Armstrong DG, Lavery LA, Diabetic Foot Study Consortium. Negative pressure wound therapy after partial diabetic foot amputation: a multicentre, randomised controlled trial[J]. Lancet, 2005, 366(9498):1704-1710.
[7]	Sajid MT, Mustafa Qu, Shaheen N, et al. Comparison of Negative Pressure Wound Therapy Using Vacuum-Assisted Closure with Advanced MoistWound Therapy in the Treatment of Diabetic Foot Ulcers[J]. J Coll Physicians Surg Pak, 2015, 25(11):789-793.
[8]	Dumville JC, Hinchliffe RJ, Cullum N, et al. Negative pressure wound therapy for treating foot wounds in people with diabetes mellitus[J]. Cochrane Database Syst Rev, 2013, 17(10):CD010318.
[9]	Zhang J, Hu ZC, Chen D, et al. Effectiveness and safety of negative-pressure wound therapy for diabetic foot ulcers: a meta-analysis[J]. Plast Reconstr Surg, 2014, 134(1):141-151.
[10]	Jung JA, Yoo KH, Han SK, et al. Influence of Negative-Pressure Wound Therapy on Tissue Oxygenation in Diabetic Feet[J]. Adv Skin Wound Care, 2016, 29(8):364-370.
[11]	Shon YS, Lee YN, Jeong SH, et al. Influence of negative-pressure wound therapy on tissue oxygenation of the foot[J]. Arch Plast Surg, 2014, 41(6):668-672.
[12]	杨少玲，孙蕾蕾，胡丽叶，等. 创面负压治疗对糖尿病足肉芽组织碱性成纤维细胞生长因子的影响[J]. 中华糖尿病杂志，2016, 8(2):103-107.
[13]	Falanga. Wound healing and its impairment in the diabetic foot[J]. Lancet, 2005, 366(9498):1736-1743.
[14]	Janis JE, Harrison B. Wound healing: part I. Basic science[J]. Plast Reconstr Surg, 2014, 133(2):199e-207e.
[15]	Reinke JM, Sorg H. Wound repair and regeneration[J]. Eur Surg Res, 2012, 49(1):35-43.
[16]	Gary Sibbald R, Woo KY. The biology of chronic foot ulcers in persons with diabetes[J]. Diabetes Metab Res Rev, 2008, 24(Suppl 1):S25-S30.
[17]	Demidova-Rice TN, Hamblin MR, Herman IM. Acute and Impaired Wound Healing: Pathophysiology and Current Methods for DrugDelivery, Part1: Normal and Chronic Wounds: Biology, Causes, and Approaches to Care[J]. Adv Skin Wound Care, 2012, 25(7):304-314.
[18]	万明才，冯峰，王新卫，等. 生肌液复合封闭负压引流技术对创面Ⅰ型和Ⅲ型胶原代谢的影响[J]. 中国组织工程研究与临床康复，2008, 12(33):6492-6495.

[1]	王杰, 袁泉, 王玥琦, 乔佳君, 谭春丽, 夏仲元, 刘守尧. 溃疡油在糖尿病足溃疡治疗中的应用效果及安全性观察[J/OL]. 中华损伤与修复杂志(电子版), 2024, 19(06): 480-484.
[2]	别瑶, 曹志斌, 辛静, 王健楠, 惠宗光. 应用基质血管成分细胞治疗糖尿病足溃疡的研究进展[J/OL]. 中华损伤与修复杂志(电子版), 2024, 19(05): 453-456.
[3]	姜珊, 李湘燕, 田硕涵, 温冰, 何睿, 齐心. 采用优化抗感染治疗模式改善糖尿病足感染预后的临床观察[J/OL]. 中华损伤与修复杂志(电子版), 2024, 19(05): 398-403.
[4]	谢芬, 陈洁, 张媛媛, 刘茜, 胡芬, 李恭驰, 李炳辉, 金环. 移动健康管理模式在糖尿病足管理中的应用效果观察[J/OL]. 中华损伤与修复杂志(电子版), 2024, 19(04): 335-340.
[5]	唐亦骁, 陈峻, 连正星, 胡海涛, 鲁迪, 徐骁, 卫强. 白果内酯对小鼠肝缺血再灌注损伤保护作用研究[J/OL]. 中华移植杂志(电子版), 2024, 18(05): 278-282.
[6]	王雪玲, 曹欢, 顾劲扬. 肠道菌群在器官缺血再灌注损伤中的作用及机制研究进展[J/OL]. 中华移植杂志(电子版), 2024, 18(04): 247-250.
[7]	孙璐, 蒋亚玲, 陈凌君. 布托啡诺对脑缺血再灌注损伤大鼠神经炎症和JAK2/STAT3信号通路的影响[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(06): 344-350.
[8]	中华医学会器官移植学分会. 肝移植术后缺血性胆道病变诊断与治疗中国实践指南[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(06): 739-748.
[9]	李京, 牛博, 刘晓蓓, 魏新雪, 黄荣. circ-SESN2 沉默靶向调控miRNA-23a-5p/ULK1 在神经细胞氧化应激损伤中的作用机制研究[J/OL]. 中华神经创伤外科电子杂志, 2024, 10(05): 263-272.
[10]	吉莉, 苏云楠, 王斌, 沈滔, 刘团结, 毛蕾, 徐玉萍, 张婷, 王博. 急性缺血性脑卒中患者脑白质微结构改变对长期认知功能损伤的预测价值研究[J/OL]. 中华脑科疾病与康复杂志(电子版), 2024, 14(04): 193-200.
[11]	张琛, 秦鸣, 董娟, 陈玉龙. 超声检查对儿童肠扭转缺血性改变的诊断价值[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(06): 565-568.
[12]	贾玲玲, 滕飞, 常键, 黄福, 刘剑萍. 心肺康复在各种疾病中应用的研究进展[J/OL]. 中华临床医师杂志(电子版), 2024, 18(09): 859-862.
[13]	梁铭垚, 袁健瑜, 关雨霏, 左思程, 阳新明, 陈巧燕. 多壳扩散磁共振成像在缺血性卒中模型的应用[J/OL]. 中华介入放射学电子杂志, 2024, 12(04): 362-368.
[14]	耿晓坤. 缺血性卒中后无效再灌注的时间窗、组织窗与神经保护[J/OL]. 中华脑血管病杂志(电子版), 2024, 18(06): 636-636.
[15]	唐欣, 翟文海, 王润婷, 周胜宇, 靳航. 补体在缺血性卒中疾病中的研究进展[J/OL]. 中华脑血管病杂志(电子版), 2024, 18(04): 382-392.

阅读次数

全文

摘要

选择文件类型/文献管理软件名称

选择包含的内容

Effects of negative pressure wound therapy on the expression of typeⅠcollagen in granulation tissue of different ischemic wound of diabetic foot

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

Effects of negative pressure wound therapy on the expression of typeⅠcollagen in granulation tissue of different ischemic wound of diabetic foot