负压伤口治疗对糖尿病足创面肉芽组织血小板源生长因子表达的影响

doi:10.3877/cma.j.issn.1673-9450.2016.02.006

中华损伤与修复杂志(电子版) ›› 2016, Vol. 11 ›› Issue (02) : 107 -112. doi: 10.3877/cma.j.issn.1673-9450.2016.02.006

所属专题：文献；

论著

负压伤口治疗对糖尿病足创面肉芽组织血小板源生长因子表达的影响

杨少玲¹, 孙蕾蕾², 胡丽叶¹, 李晓玲¹, 杨轶文¹, 李艳俊¹, 朱旅云¹^,^†(

)

^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

收稿日期:2015-11-08 出版日期:2016-04-01
通信作者: 朱旅云
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)

Effect of negative pressure wound therapy on the expression of platelet derived growth factor in granulation tissue of diabetic foot wounds

Shaoling Yang¹, Leilei Sun², Liye Hu¹, Xiaoling Li¹, Yiwen Yang¹, Yanjun Li¹, Lyuyun Zhu¹^,^†()

^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

Received:2015-11-08 Published:2016-04-01
Corresponding author: Lyuyun Zhu
About author:
Corresponding author: Zhu Lyuyun, Email: yshling9@126.com

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引用本文：

杨少玲, 孙蕾蕾, 胡丽叶, 李晓玲, 杨轶文, 李艳俊, 朱旅云. 负压伤口治疗对糖尿病足创面肉芽组织血小板源生长因子表达的影响[J/OL]. 中华损伤与修复杂志(电子版), 2016, 11(02): 107-112.

Shaoling Yang, Leilei Sun, Liye Hu, Xiaoling Li, Yiwen Yang, Yanjun Li, Lyuyun Zhu. Effect of negative pressure wound therapy on the expression of platelet derived growth factor in granulation tissue of diabetic foot wounds[J/OL]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2016, 11(02): 107-112.

目的

探讨负压伤口治疗技术(NPWT)的应用对糖尿病足肉芽组织血小板源生长因子(PDGF)表达的影响。

方法

选取解放军白求恩国际和平医院2010年1月至2014年12月收治的40例糖尿病足患者，采用单纯随机抽样法将患者随机分为NPWT组(20例)和对照组(20例)，NPWT组创面采用负压伤口治疗技术治疗，对照组采用传统纱布换药治疗。两组分别于治疗前及治疗第12天取创面肉芽组织2份，一份置于4%多聚甲醛溶液中保存，行苏木精－伊红(HE)染色和免疫组织化学染色，HE染色观察肉芽组织形成情况，免疫组织化学染色观察肉芽组织PDGF的表达情况，通过Motic Medical 6.0数码医学图像分析系统检测吸光度值对PDGF蛋白行定量分析。同时，另一份肉芽组织标本－80℃冰箱中保存，采用Western blot法检测PDGF蛋白表达量。数据应用SPSS 13.0软件进行分析，PDGF吸光度值及PDGF蛋白表达量比较采用两独立样本t检验。

结果

两组治疗前HE染色可见炎症细胞多，成纤维细胞、新生血管形成较少；两组治疗第12天肉芽组织HE染色均可见新鲜肉芽组织的形成，表现为新生血管、成纤维细胞增多，但与对照组相比，NPWT组新生肉芽组织增多较为明显。免疫组织化学分析显示，两组治疗后均可见PDGF蛋白表达上调；NPWT组治疗第12天PDGF吸光度值(0.41±0.05)与治疗前(0.29±0.04)比较，差异有统计学意义(t＝8.63，P<0.05)；对照组治疗第12天PDGF吸光度值(0.33±0.04)与治疗前(0.28±0.04)比较，差异也有统计学意义(t＝3.62，P<0.05)；但是，NPWT组治疗前后PDGF吸光度变化值为0.13±0.05，与对照组治疗前后PDGF吸光度变化值(0.05±0.03)比较，差异有统计学意义(t＝5.89，P<0.05)。Western blot分析显示，NPWT组治疗第12天PDGF蛋白表达量为0.24 ±0.04，与治疗前PDGF蛋白表达量(0.11±0.03)比较，差异有统计学意义(t＝11.01，P<0.05)；对照组治疗第12天PDGF蛋白表达量为0.17±0.03，与治疗前PDGF蛋白表达量(0.12 ±0.03)比较，差异亦有统计学意义(t＝6.06，P<0.05)；但是，NPWT组治疗前后PDGF蛋白表达量的变化值为0.12 ±0.03，与对照组治疗前后PDGF蛋白表达量的变化值(0.05 ±0.02)比较，差异有统计学意义(t＝9.06，P<0.05)。

结论

应用NPWT可明显促进创面肉芽组织PDGF表达上调，从而促进创面的愈合。

关键词: 负压伤口疗法, 糖尿病足, 受体，血小板源生长因子

Objective

To assess the endogenous regulation of platelet derived growth factor (PDGF) in the foot wound healing response induced by negative pressure wound therapy (NPWT) in human diabetic patients in vivo.

Methods

From January 2010 to December 2014, 40 patients with diabetic foot wounds fitting the inclusion criteria were found and enrolled in the clinical trial. The subjects were randomly assigned for NPWT group or control group with simple random sampling method. Twenty patients were treated with NPWT, while the same number of patients in the control group received conventional gauze therapy. Granulated tissue biopsies were collected before(0 day) and after(12 days) treatment in both groups. All biopsies were subdivided into two parts. One part was preserved in 4% paraformaldehyde for hematoxylin eosin staining and immunocytochemical staining of PDGF, the other part was stored at -80℃ for Western blot analysis of PDGF expression. Experimental data were analyzed using SPSS 13.0 software and differences of absorbance value and protein expression of PDGF between the two groups were analyzed using two-sample t test.

Results

The HE staining showed that both NPWT group and control group wounds developed characteristic granulated tissue after treatment, which presented intense fibroblasts and neoformation of vessels. However, NPWT group wounds developed much more characteristic granulated tissue than control group wounds. The immunohistochemical data revealed that PDGF expression was up-regulated in NPWT group and control group after treatment. The absorbance value of PDGF was higher after treatment than that before treatment in NPWT group, the difference was statistically significant (0.41±0.05 vs 0.29±0.04, t=8.63, P<0.05). Also, the absorbance value of PDGF was higher after treatment than that before treatment in control group, the difference was statistically significant (0.33±0.04 vs 0.28±0.04, t=3.62, P<0.05). However, the change value (difference between before treatment and after treatment) of absorbance value of PDGF was higher in NPWT group than that in control group, the difference was statistically significant (0.13±0.05 vs 0.05±0.03, t=5.89, P<0.05). The Western blot analysis of PDGF was consistent with that of immunohistochemical analysis. The protein levels of PDGF was higher after treatment than that before treatment in NPWT group, the difference was statistically significant (0.24 ±0.04 vs 0.11±0.03, t=11.01, P<0.05). Also, the protein level of PDGF was higher after treatment than that before treatment in control group, the difference was statistically significant (0.17±0.03 vs 0.12±0.03, t=6.06, P<0.05). But, the change value ( difference between before treatment and after treatment) of protein level of PDGF was higher in NPWT group than that in control group, the difference was statistically significant (0.12 ±0.03 vs 0.05 ±0.02, t=9.06, P<0.05).

Conclusion

NPWT obviously enhances PDGF expression of granulation tissue in diabetic foot wound and subsequently promotes wound healing.

Key words: Negative-pressure wound therapy, Diabetic foot, Receptors, platelet derived growth factor

表1 糖尿病患者人口统计学数据和实验前各观察指标(n＝20)

患者一般情况	对照组	NPWT组	检验值	P值
年龄(岁)	59.20±6.44	59.05±6.77	0.07	0.94
性别(男/女)	15/5	10/10	2.67^a	0.10
ABI	0.63±0.09	0.64±0.08	0.49	0.63
TcPO₂(mmHg)	28.80±5.76	29.30±4.67	0.30	0.77
FBG(mmol/L)	8.13±3.84	10.29±3.53	1.86	0.07
HbA1c (%)	10.24±1.96	9.80±2.05	0.69	0.50
溃疡时间(d)	54.25±18.22	58.30±4.57	0.66	0.51
肌酐(μmol/L)	55.50(45.75，71.50)	65(42.25，95.00)	0.87^b	0.40
转氨酶(U/L)	11.95(9.00，19.25)	14(10.00，21.88)	0.66^b	0.51
合并神经病变例数	18	19	0.28^a	0.60

表1 糖尿病患者人口统计学数据和实验前各观察指标(n＝20)

患者一般情况	对照组	NPWT组	检验值	P值
年龄(岁)	59.20±6.44	59.05±6.77	0.07	0.94
性别(男/女)	15/5	10/10	2.67^a	0.10
ABI	0.63±0.09	0.64±0.08	0.49	0.63
TcPO₂(mmHg)	28.80±5.76	29.30±4.67	0.30	0.77
FBG(mmol/L)	8.13±3.84	10.29±3.53	1.86	0.07
HbA1c (%)	10.24±1.96	9.80±2.05	0.69	0.50
溃疡时间(d)	54.25±18.22	58.30±4.57	0.66	0.51
肌酐(μmol/L)	55.50(45.75，71.50)	65(42.25，95.00)	0.87^b	0.40
转氨酶(U/L)	11.95(9.00，19.25)	14(10.00，21.88)	0.66^b	0.51
合并神经病变例数	18	19	0.28^a	0.60

图1 NPWT组与对照组患者治疗前和治疗第12天伤口肉芽组织染色结果分析(苏木精－伊红染色×400)

图2 NPWT组与对照组治疗前和治疗第12天糖尿病足创面肉芽组织PDGF蛋白表达情况(免疫组织化学染色×400)

图3 NPWT组与对照组糖尿病足创面肉芽组织血小板源生长因子吸光度值检测结果

图4 Western blot法检测NPWT组和对照组治疗前和治疗第12天糖尿病足创面肉芽组织PDGF蛋白表达情况

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