【摘要】 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。更多还原

【Abstract】 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 had15 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 type Ⅰ collagen 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 typeⅠ collagen 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 type Ⅰ collagen 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.更多还原