高压氧治疗对2型糖尿病足大鼠模型皮肤组织中血红素加氧酶-1表达的影响

Effects of hyperbaric oxygen on the expression of HO-1 in the dermal tissue of rats with type Ⅱ diabetic foot

目的 探讨高压氧治疗对血红素加氧酶-1（HO-1）在糖尿病足大鼠皮肤组织中表达的影响以及HO-1水平与溃疡愈合的相关性.方法 健康成年雄性Wistar大鼠45只,体质量250～350 g,按数字表法随机分为糖尿病足部溃疡组（溃疡组）、糖尿病足部溃疡高压氧治疗组（高压氧组）、正常对照组（对照组）,每组15只.3组均以高脂饲料（88.8％基础饲料、1％胆固醇、10％猪油、0.2％胆盐）饲养1个月,制作成2型糖尿病模型,溃疡组和高压氧组大鼠再制作成2型糖尿病足溃疡模型,高压氧组用高压氧治疗（治疗压力0.2 MPa,1次/d,连续治疗15 d）.然后取3组大鼠足部皮肤,采用逆转录聚合酶链反应（PT-PCR）检测皮肤组织中HO-1 mRNA的表达;采用免疫组织化学方法检测HO-1蛋白的表达.结果 溃疡组、高压氧组、对照组HO-1 mRNA表达量分别为1.063±0.006、1.082±0.007、0.950±0.013,溃疡组、高压氧组与对照组比较,溃疡组与高压氧组比较,差异均有统计学意义（P＜0.01）.免疫组织化学结果显示：对照组HO-1表达主要集中在表皮组织;溃疡组和高压氧组表皮和真皮组织中的HO-1表达较正常对照组明显增加,且高压氧组HO-1表达都多于溃疡组.结论 糖尿病足部溃疡与其皮肤组织中的HO-1表达呈相关性,糖尿病足部溃疡皮肤组织的抗氧化能力降低.

Objective To explore the expression of HO-1 in the dermal tissue of rats with type Ⅱ diabetic foot ulcer and the association between HO-1 level and ulcer healing.Methods Forty-five Wistar male healthy rats with the weight ranging from 250 to 350 g were randomly divided into 3 groups：the diabetic foot ulcer group（the DF group）,the diabetic foot ulcer hyperbaric oxygen therapy group （or the DFHBO group or simply the HBO group） and the normal control group （or the NC group）,each consisting of 15 animals.The mRNA transcription of HO-1 was detected by RT-PCR.The animals of the 3 groups were all raised with high lipid feed （88.8％ basic feed,1％ cholesterol,10％ lard and 0.2％ bile salt） for a period of one month.Then,the animal model of type Ⅱ diabetes was developed,and the model of type Ⅱ diabetes with diabetic food ulcer was further developed in the animals of the DF group and the DFHBO group.The animals in the DFHBO group were treated with HBO,at a pressure ofO.2 MPa,one session a day,for a duration of 15 days.Then,skin samples were collected from the foot of the animals in the 3 groups.The expression of HO-1 mRNA was detected by PT-PCR and the expression of HO-1 was determined by immunohistochemistry.Results The mRNA expression levels in the DF group,the HBO group and the NC group were respectively 1.063 ± 0.006,1.082±0.007 and 0.950 ± 0.013.Statistical significance could be noted,when comparisons were made between the 3 groups （P ＜0.01）,and statistical difference could also be seen,when comparisons were made between the DF group and the HBO group （P ＜ 0.01）.Immunohistochemistry detection indicated that HO-1 expression was more significant in epidermal tissue for the NC group.The expression levels of HO-1 both in the epidermal and dermal tissues for the animals of the DF group and the HBO group were more significant,when comparisons were made with that of the NC group,and the HO-1 expression level for the animals in the HBO group was higher than that of the DF group.Conclusions Diabetic foot ulcer was positively correlated with the expression of HO-1 in the dermal tissue.It was possible that antioxidation decreased in the dermal tissue of rats with diabetic foot ulcers.