MEBT/MEBO对糖尿病大鼠创面组织中ULK1表达的影响

目的观察皮肤再生医疗技术(moist exposed burn therapy/moist exposed burn ointment,MEBT/MEBO)对糖尿病大鼠创面组织中丝氨酸/苏氨酸蛋白激酶-失调51样激酶1(uncoordinated 51 like kinase-1,ULK1)表达的影响,探讨MEBT/MEBO促进糖尿病创面愈合的可能机制。方法选取SPF级Wistar雄性大鼠90只,随机分为对照组、模型组和MEBO组,每组30只,其中对照组大鼠建立常规创面模型,模型组和MEBO组大鼠建立糖尿病创面模型。模型建立后,对照组及模型组大鼠创面予以生理盐水纱布外敷治疗,MEBO组大鼠创面予以湿润烧伤膏(moist exposed burn ointment,MEBO)药纱外敷治疗。分别于治疗第1、11、18天采用Western blotting法检测ULK1蛋白及qRT-PCR技术检测ULK1 mRNA的表达水平,并对比观察3组大鼠创面愈合情况。结果(1)治疗第1天,3组大鼠创面均淤血水肿,无肉芽组织形成;治疗第11天,对照组大鼠创面干燥,可见大片肉芽组织形成,模型组大鼠创面高度水肿,无肉芽组织形成,MEBO组大鼠创面轻微水肿,可见散在肉芽组织形成;治疗第18天,对照组大鼠创面完全愈合,模型组大鼠创面轻微水肿,可见散在肉芽组织形成,MEBO组大鼠创面干燥,可见大片肉芽组织形成。(2)治疗第11、18天,3组大鼠创面愈合率对比,对照组>MEBO组>模型组,组间两两对比,P均<0.05,差异具有统计学意义。(3)治疗第1天,3组大鼠创面组织中ULK1蛋白及ULK1 mRNA表达水平对比,对照组>MEBO组=模型组,对照组与MEBO组及对照组与模型组对比,P均<0.05,差异具有统计学意义;治疗第11天,3组大鼠创面组织中ULK1蛋白及ULK1 mRNA表达水平对比,对照组>MEBO组>模型组,组间两两对比,P均<0.05,差异具有统计学意义;治疗第18天,3组大鼠创面组织中ULK1蛋白及ULK1 mRNA表达水平对比,对照组=MEBO组<模型组,对照组与模型组及MEBO组与模型组对比,P均<0.05,差异具有统计学意义。结论MEBT/MEBO可有效促进糖尿病大鼠创面愈合,提高创面组织中ULK1表达水平,诱导细胞自噬可能是其作用机制之一。

Effect of MEBT/MEBO on the Expression of ULK1 in Wound Tissues of Rats with Diabetes

Objective To observe the effect of Moist Exposed Burn Therapy/Moist Exposed Burn Ointment(MEBT/MEBO)on the expression of serine/threonine protein kinase?uncoordinated 51 like kinase?1 in wound tissues of rats with diabetes,and explore the possible mechanism of action of MEBT/MEBO in promoting the healing of diabetic wounds.Methods Ninety SPF Wistar male rats were randomly divided into a control group,a model group and a MEBO group with 30 rats in each group.Conventional wound models were established in the control group while diabetic wound models were respectively established in the model group and MEBO group.After the models were established,the wounds in the control group and model group were treated with the dressing change of normal saline?impregnated gauze,while the wounds in the MEBO group were treated with the dressing change of MEBO?impregnated gauze.The expression levels of ULK1 protein and ULK1 mRNA were respectively tested by Western blotting and qRT?PCR on day 1,11 and 18 of treat?ment,and the wound healing condition was observed in the three groups.Results(1)On day 1 of treatment,congestion and edema were observable in the wounds of the three groups,without the formation of granulation tissues.On day 11 of treatment,the wounds in the control group were dry with large pieces of granulation tissues visible;the wounds in the model group were highly edematous without the formation of granulation tissues;while the wounds in the MEBO group were mildly edematous with scattered granulation tissues observable.On day 18 of treatment,the wounds in the control group were com?pletely healed,the wounds in the model group were mildly edematous with scattered granulation tissues visible,while the wounds in the MEBO group were dry with large pieces of granulation tissue formation.(2)On day 11 and 18 of treatment,the wound healing rate was compared among the three groups and the results showed as control group>MEBO group>model group.The pairwise comparisons of the wound healing rate among the three groups all showed statistically significant differ?ences(P<0??05).(3)The expression levels of ULK1 protein and ULK1 mRNA in the wound tissues of the three groups were compared respectively on day 1,11 and 18 of treatment and the results were respectively control group>MEBO group=model group,control group>MEBO group>model group,and control group=MEBO group