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]. 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]. 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]	张健, 刘小龙, 查天建, 姚俊杰, 王傑. 富含血小板血浆联合异种脱细胞真皮基质修复糖尿病足缺血性创面的临床效果[J]. 中华损伤与修复杂志(电子版), 2023, 18(06): 503-506.
[2]	赵雅玫, 谢斌, 陈艳, 吴健. 抗生素骨水泥联合负压封闭引流对糖尿病足溃疡临床疗效的荟萃分析[J]. 中华损伤与修复杂志(电子版), 2023, 18(05): 427-433.
[3]	鲍亚慧, 曹志斌, 王健楠, 别瑶, 孙晓东, 惠宗光. 应用羧甲基纤维素钠银敷料联合封闭负压吸引治疗糖尿病足溃疡的疗效[J]. 中华损伤与修复杂志(电子版), 2023, 18(04): 326-330.
[4]	张升敏, 黄健斌, 陈亮, 马克强. Ⅰ、Ⅲ型胶原蛋白在成人腹股沟斜疝及直疝患者腹横筋膜和疝囊的表达[J]. 中华疝和腹壁外科杂志(电子版), 2023, 17(05): 516-521.
[5]	关明函, 薛志强. 右美托咪定改善大鼠脑缺血再灌注后脑损伤的研究[J]. 中华神经创伤外科电子杂志, 2023, 09(05): 270-276.
[6]	何彬, 王静. 彩色多普勒超声血流参数、血清尿酸、胱抑素C对短暂性脑缺血发作患者颈动脉狭窄的诊断价值[J]. 中华神经创伤外科电子杂志, 2023, 09(05): 289-294.
[7]	尚慧娟, 袁晓冬. 机械取栓术后应用依达拉奉右崁醇对急性缺血性脑卒中预后的改善[J]. 中华神经创伤外科电子杂志, 2023, 09(05): 295-301.
[8]	王静, 何彬. 经颅彩色多普勒超声联合血sCD40L、Fib、PAF对短暂性脑缺血发作后脑梗死的预测效果[J]. 中华神经创伤外科电子杂志, 2023, 09(04): 222-227.
[9]	李变, 王莉娜, 桑田, 李珊, 杜雪燕, 李春华, 张兴云, 管巧, 王颖, 冯琪, 蒙景雯. 亚低温技术治疗缺氧缺血性脑病新生儿的临床分析[J]. 中华临床医师杂志(电子版), 2023, 17(06): 639-643.
[10]	张敏洁, 张小杉, 段莎莎, 施依璐, 赵捷, 白天昊, 王雅晳. 氢气治疗心肌缺血再灌注损伤的作用机制及展望[J]. 中华临床医师杂志(电子版), 2023, 17(06): 744-748.
[11]	张敏洁, 王雅晳, 段莎莎, 施依璐, 付文艳, 赵海玥, 张小杉. 基于GEO数据库和生物信息学分析筛选大鼠心肌缺血再灌注损伤相关潜在通路和靶点[J]. 中华临床医师杂志(电子版), 2023, 17(04): 438-445.
[12]	赵守法, 王舒, Larissa Miare Vados, Alberto Cantidio Ferreira. 醒脑开窍针刺法结合康复疗法治疗缺血性中风病恢复期的疗效观察[J]. 中华针灸电子杂志, 2023, 12(04): 151-155.
[13]	张许平, 刘佳成, 张舸, 杜艳姣, 李韶, 商丹丹, 王浩, 李艳, 段智慧. CYP2C19基因多态性联合血栓弹力图指导大动脉粥样硬化型非致残性缺血性脑血管事件患者抗血小板治疗的效果[J]. 中华脑血管病杂志(电子版), 2023, 17(05): 477-481.
[14]	金泽平, 董晶, 柳云鹏, 汪阳. 菌群-肠道-脑轴与缺血性卒中危险因素关系的研究进展[J]. 中华脑血管病杂志(电子版), 2023, 17(05): 510-517.
[15]	尹琛俊, 张喆, 李晓明. 卵圆孔未闭相关血栓形成机制的研究进展[J]. 中华脑血管病杂志(电子版), 2023, 17(04): 307-311.

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