硼酸消毒液促进糖尿病难愈性创面愈合的护理效用及相关机制初探
发布时间：2018-09-14 11:39
【摘要】：背景和目的： 糖尿病难愈性创面是包括糖尿病足和其他糖尿病创面在内的，由多种原因引起的经久不愈的创面，是最严重、花费最高的糖尿病慢性并发症之一，严重的糖尿病足可导致截肢、甚至死亡。调查显示，我国约有30%的非创伤性截肢是由于糖尿病足引发。糖尿病难愈性创面发生发展机制及护理是临床研究的难点和热点。 TGF-β1/Smads信号转导通路是糖尿病难愈性创面修复中重要的信号转导通路之一。TGF-β1是介导组织硬化和间质纤维化的关键细胞因子，参与了细胞分化、增殖、粘附、血管生成、炎症反应和创面愈合等过程，还能促成成纤维细胞的迁移和增殖，趋化、刺激成纤维细胞生成细胞外基质。研究显示，糖尿病难愈性创面的形成与创面TGF-β1表达的绝对或相对不足有关。近年，弱酸环境对难愈性创面的愈合有促进作用的临床个案已有少量报道。我们认为其机制除了弱酸环境对创面微生物有一定的抑制作用外，还可能与其能刺激创面Tgf-β1mRNA的表达有关，这可能是其促进创面愈合的重要机制之一，值得深入研究。 4%硼酸溶液作为临床常见的消毒剂，具有弱酸性和较低的细胞毒性，多用于皮肤、粘膜及伤口的消毒处置，，有部分临床研究也曾运用硼酸对下肢浅表溃疡、褥疮等创面进行处理并取得了较好效果。本研究因此进行了硼酸消毒液促进糖尿病难愈性创面愈合的护理效用及其相关机制的实验研究，以期为临床糖尿病难愈性创面护理措施的细化和修订提供更科学的参考依据。 材料与方法： 1.动物分组与模型建立：8周龄雄性C57BL/6小鼠30只，购买并饲养于第三军医大学实验动物中心SPF级动物实验室，随机分为糖尿病组与正常对照组，各15只。每组中又分为3天、7天、14天三个时相点（每个时相点5只）。糖尿病组小鼠以链脲佐菌素诱导建模，角膜环钻制作小鼠背部创面。 2.每日1次，在同一时段分别以0.9%生理盐水（A处理）、4%硼酸溶液（B处理）、0.5%碘伏+3%过氧化氢溶液（C处理）对不同组别小鼠创面进行清洗换药，观察记录创面结痂情况、愈合情况及创面大小，并拍照。 3.在不同时相点用Trizol提取小鼠创面组织的总RNA，在检测RNA浓度、RT-PCR反应后，获得cDNA。 4.采用SYBR Green Q-PCR法对上述创面组织的cDNA进行Tgf-β1、Smad4、Collagen I的mRNA相对表达量检测。 结果： 1.硼酸对创面愈合率的影响 糖尿病硼酸处理组各时相点创面愈合率较其余处理组的创面愈合率提高明显（P＜0.05）。 2.硼酸对创面Tgf-β1mRNA表达的影响 正常小鼠硼酸处理组3天时Tgf-β1mRNA的表达均较其余处理组明显增高（P＜0.05）；糖尿病小鼠硼酸处理组在伤后3天、14天时其Tgf-β1mRNA的表达较其余处理组有显著升高（P＜0.01）。 3.硼酸对创面Smad4mRNA表达的影响 正常小鼠硼酸处理组伤后14天时其Smad4mRNA表达低于其余处理组（P＜0.05）；糖尿病小鼠伤后14天生理盐水组Smad4mRNA的表达明显低于碘伏+双氧水处理组（P＜0.05）。 4.硼酸对创面Collagen I mRNA表达的影响 糖尿病小鼠和正常小鼠在伤后3天时均表现为硼酸处理组和碘伏+双氧水处理组的Collagen I mRNA的表达高于生理盐水组（P＜0.05）；14天时，糖尿病小鼠硼酸组和生理盐水组Collagen I mRNA的表达高于碘伏＋双氧水处理组（P＜0.05），正常小鼠则是碘伏+双氧水组Collagen I mRNA表达高于生理盐水和硼酸组。 结论： 1.不管在早期还是中晚期，硼酸处理对提高糖尿病小鼠伤口愈合率，促进创面愈合均达到了很好的效果； 2.硼酸促进糖尿病小鼠创面愈合的作用机制可能与其特殊的酸性条件能更好地刺激创面Tgf-β1、Collagen I mRNAs的表达有关； 3.正常小鼠早期使用硼酸和生理盐水都能较好地促进创面愈合，晚期使用碘伏+双氧水对促进创面愈合效果更好。
[Abstract]:Background and purpose:
Diabetic refractory wounds include diabetic foot and other diabetic wounds. Prolonged wounds caused by various causes are one of the most serious and costly chronic complications of diabetes. Severe diabetic foot can lead to amputation or even death. The pathogenesis and nursing of diabetic refractory wounds is a difficult and hot topic in clinical research.
TGF-beta 1/Smads signal transduction pathway is one of the important signal transduction pathways in repairing diabetic refractory wounds. Proliferation, chemotaxis, and stimulation of fibroblasts to produce extracellular matrix. Studies have shown that the formation of refractory diabetic wounds is related to the absolute or relative inadequacy of TGF-beta 1 expression in the wounds. In recent years, a few clinical cases have reported that weak acid environment can promote the healing of refractory wounds. In addition to biological inhibition, it may also be related to its possible stimulation of Tgf-beta 1 mRNA expression in wounds, which may be one of the important mechanisms of promoting wound healing and worthy of further study.
4% boric acid solution as a common clinical disinfectant, with weak acidity and low cytotoxicity, is often used for disinfection of skin, mucosa and wounds. Some clinical studies have also used boric acid on lower extremity ulcer, bedsore and other wounds and achieved good results. The experimental study on the nursing effect and related mechanism of refractory wound healing is expected to provide a more scientific reference for the refinement and revision of nursing measures for refractory wounds of diabetes mellitus.
Materials and methods:
1. Animal grouping and model establishment: 30 8-week-old male C57BL/6 mice were purchased and fed in SPF class Animal Laboratory of Experimental Animal Center of Third Military Medical University. They were randomly divided into diabetes group and normal control group, 15 mice in each group. Induced modeling, the cornea ring drill made mouse back wound.
2. The wounds of mice in different groups were cleaned and dressed once a day with 0.9% normal saline (A), 4% boric acid (B) and 0.5% iodophor + 3% hydrogen peroxide (C), respectively. The scab formation, healing and wound size were observed and recorded.
3. Trizol was used to extract the total RNA from the wound tissue of mice at different time points. After detecting the concentration of RNA and RT-PCR reaction, the cDNA was obtained.
4. The relative expression of Tgf-beta 1, Smad4 and Collagen I in the above wound tissues was detected by SYBR Green Q-PCR.
Result:
Effect of 1. boric acid on wound healing rate
The wound healing rate of diabetic boric acid treatment group was higher than that of other treatment groups at each time point (P < 0.05).
Effect of 2. boric acid on the expression of Tgf- beta 1mRNA in the wound
The expression of Tgf-beta 1 mRNA in the boric acid treated group was significantly higher than that in the other groups at 3 days after injury (P < 0.05), and the expression of Tgf-beta 1 mRNA in the boric acid treated group was significantly higher than that in the other groups at 3 days and 14 days after injury (P < 0.01).
Effect of 3. boric acid on Smad4mRNA expression in wound surface
Smad4 mRNA expression in normal mice treated with boric acid at 14 days after injury was lower than that in other treatment groups (P < 0.05); Smad4 mRNA expression in normal saline group at 14 days after injury was lower than that in iodophor + hydrogen peroxide treatment group (P < 0.05).
Effect of 4. boric acid on the expression of Collagen I mRNA in wound
The expression of Collagen I mRNA in boric acid treatment group and iodophor + hydrogen peroxide treatment group was higher than that in normal saline group at 3 days after injury (P < 0.05); at 14 days, the expression of Collagen I mRNA in boric acid group and normal saline group was higher than that in iodophor + hydrogen peroxide treatment group (P < 0.05). The expression of Collagen I mRNA in iodophor + H2O2 group was higher than that in normal saline and boric acid group.
Conclusion:
1. Boric acid treatment can improve the wound healing rate and promote the wound healing of diabetic mice in the early and middle and late stages.
2. The mechanism of the effect of boric acid on wound healing in diabetic mice may be related to the special acidic condition which can better stimulate the expression of Tgf-beta 1 and Collagen I mRNAs.
3. Early use of boric acid and normal saline can promote wound healing in normal mice, and late use of iodophor and hydrogen peroxide can promote wound healing better.
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R473.5
本文编号：2242607
[Abstract]:Background and purpose:
Diabetic refractory wounds include diabetic foot and other diabetic wounds. Prolonged wounds caused by various causes are one of the most serious and costly chronic complications of diabetes. Severe diabetic foot can lead to amputation or even death. The pathogenesis and nursing of diabetic refractory wounds is a difficult and hot topic in clinical research.
TGF-beta 1/Smads signal transduction pathway is one of the important signal transduction pathways in repairing diabetic refractory wounds. Proliferation, chemotaxis, and stimulation of fibroblasts to produce extracellular matrix. Studies have shown that the formation of refractory diabetic wounds is related to the absolute or relative inadequacy of TGF-beta 1 expression in the wounds. In recent years, a few clinical cases have reported that weak acid environment can promote the healing of refractory wounds. In addition to biological inhibition, it may also be related to its possible stimulation of Tgf-beta 1 mRNA expression in wounds, which may be one of the important mechanisms of promoting wound healing and worthy of further study.
4% boric acid solution as a common clinical disinfectant, with weak acidity and low cytotoxicity, is often used for disinfection of skin, mucosa and wounds. Some clinical studies have also used boric acid on lower extremity ulcer, bedsore and other wounds and achieved good results. The experimental study on the nursing effect and related mechanism of refractory wound healing is expected to provide a more scientific reference for the refinement and revision of nursing measures for refractory wounds of diabetes mellitus.
Materials and methods:
1. Animal grouping and model establishment: 30 8-week-old male C57BL/6 mice were purchased and fed in SPF class Animal Laboratory of Experimental Animal Center of Third Military Medical University. They were randomly divided into diabetes group and normal control group, 15 mice in each group. Induced modeling, the cornea ring drill made mouse back wound.
2. The wounds of mice in different groups were cleaned and dressed once a day with 0.9% normal saline (A), 4% boric acid (B) and 0.5% iodophor + 3% hydrogen peroxide (C), respectively. The scab formation, healing and wound size were observed and recorded.
3. Trizol was used to extract the total RNA from the wound tissue of mice at different time points. After detecting the concentration of RNA and RT-PCR reaction, the cDNA was obtained.
4. The relative expression of Tgf-beta 1, Smad4 and Collagen I in the above wound tissues was detected by SYBR Green Q-PCR.
Result:
Effect of 1. boric acid on wound healing rate
The wound healing rate of diabetic boric acid treatment group was higher than that of other treatment groups at each time point (P < 0.05).
Effect of 2. boric acid on the expression of Tgf- beta 1mRNA in the wound
The expression of Tgf-beta 1 mRNA in the boric acid treated group was significantly higher than that in the other groups at 3 days after injury (P < 0.05), and the expression of Tgf-beta 1 mRNA in the boric acid treated group was significantly higher than that in the other groups at 3 days and 14 days after injury (P < 0.01).
Effect of 3. boric acid on Smad4mRNA expression in wound surface
Smad4 mRNA expression in normal mice treated with boric acid at 14 days after injury was lower than that in other treatment groups (P < 0.05); Smad4 mRNA expression in normal saline group at 14 days after injury was lower than that in iodophor + hydrogen peroxide treatment group (P < 0.05).
Effect of 4. boric acid on the expression of Collagen I mRNA in wound
The expression of Collagen I mRNA in boric acid treatment group and iodophor + hydrogen peroxide treatment group was higher than that in normal saline group at 3 days after injury (P < 0.05); at 14 days, the expression of Collagen I mRNA in boric acid group and normal saline group was higher than that in iodophor + hydrogen peroxide treatment group (P < 0.05). The expression of Collagen I mRNA in iodophor + H2O2 group was higher than that in normal saline and boric acid group.
Conclusion:
1. Boric acid treatment can improve the wound healing rate and promote the wound healing of diabetic mice in the early and middle and late stages.
2. The mechanism of the effect of boric acid on wound healing in diabetic mice may be related to the special acidic condition which can better stimulate the expression of Tgf-beta 1 and Collagen I mRNAs.
3. Early use of boric acid and normal saline can promote wound healing in normal mice, and late use of iodophor and hydrogen peroxide can promote wound healing better.
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R473.5
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