摘要
目的 探讨胸腺素β4(thymosin β4,Tβ4)联合水胶体敷料对深Ⅱ度烧伤大鼠早期创面愈合及p38丝裂原活化蛋白激酶(mitogen activated protein kinase, MAPK)信号传导通路的影响。方法 选取常规适应性喂养的健康Wister雄性大鼠40只作为研究对象,随机抽取10只作为对照组,其余30只按照随机数字表法分为烧伤组、Tβ4+水胶体敷料组、p38MAPK激动剂组,每组10只。除对照组外,其余大鼠均制作深Ⅱ度烧伤模型,造模成功后进行后续试验,Tβ4+水胶体敷料组给予外涂6μg Tβ4+安普贴贴敷,p38MAPK激动剂组给予外涂6μg Tβ4+安普贴贴敷+外涂p79350(1μg/kg),烧伤组大鼠给予等量0.9%氯化钠溶液外涂。给药3、7、10、14 d对创面进行拍照,采用MoticMed6软件及Photoshop软件处理照片并计算各组大鼠早期创面愈合率;免疫组织化学法检测创面肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素10(interleukin-10,IL-10)等炎症因子水平;逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction, RT-PCR)检测创面转化生长因子β1蛋白(transforming growth factor β1 protein, TGF-β1)mRNA、血管内皮生长因子(vascular endothelial growth factor, VEGF)mRNA表达情况;蛋白免疫印迹法(Western blotting, WB)检测p38MAPK信号传导通路关键蛋白p38MAPK、核因子κB(nuclear factorκB,NF-κB)、半胱氨酸蛋白酶3(cysteine protease 3,caspase-3)表达情况。结果 给药3、7、10、14 d后,Tβ4+水胶体敷料组的创面愈合率均高于烧伤组和p38MAPK激动剂组(P<0.05)。与对照组比较,烧伤组、Tβ4+水胶体敷料组、p38MAPK激动剂组各时点TNF-α、IL-10水平均较高(P<0.05);给药后12、24、48 h,烧伤组、p38MAPK激动剂组TNF-α水平高于Tβ4+水胶体敷料料组(P<0.05),IL-10水平低于Tβ4+水胶体敷料组(P<0.05);而烧伤组、p38MAPK激动剂组各时点TNF-α、IL-10水平差异无统计学意义(P>0.05)。与对照组比较,烧伤组、Tβ4+水胶体敷料组、p38MAPK激动剂组各时间点TGF-β1 mRNA、VEGF mRNA表达均上调(P<0.05);与烧伤组比较,Tβ4+水胶体敷料组各时间点TGF-β1 mRNA表达均下调(P<0.05)、VEGF mRNA表达均上调(P<0.05);烧伤组、p38MAPK激动剂组各时间点TGF-β1 mRNA、VEGF mRNA表达差异均无统计学意义(P>0.05)。与对照组比较,烧伤组、Tβ4+水胶体敷料组、p38MAPK激动剂组p38MAPK、NF-κB、caspase-3蛋白表达量均较高(P<0.05);与烧伤组比较,Tβ4+水胶体敷料组p38MAPK、NF-κB、caspase-3蛋白表达量均较低(P<0.05);但烧伤组、p38MAPK激动剂组p38MAPK、NF-κB、caspase-3蛋白表达量差异无统计学意义(P>0.05)。结论 胸腺素β4联合水胶体敷料有助于促进深Ⅱ度烧伤创面愈合,其机制可能与抑制p38丝裂原活化蛋白激酶信号转导通路、降低炎性因子表达有关。
Objective To investigate the effect of thymosinβ4(Tβ4)combined with hydrocolloid dressing on early wound healing and p38 mitogen-activated protein kinase(MAPK)signal transduction pathway in rats with deepⅡdegree burn.Methods Forty healthy Wister male rats with conventional adaptive feeding were selected as the research subjects.Ten rats were randomly selected as the control group,and the remaining 30 animals were divided into burn group(n=10),Tβ4+hydrocolloid dressing group(n=10)and p38MAPK agonist group(n=10)according to the random number table method.Except the control group,the remaining rats were made into deepⅡdegree burn models for subsequent experiments.The Tβ4+hydrocolloid dressing group was given external application of 6μg Tβ4+Ampu patch,the p38MAPK agonist group was given external application of 6μg Tβ4+AMP patch+external application of p79350(1μg/kg),and the burn group was given an equal amount of 0.9%sodium chloride solution for external application.The wounds were photographed on the 3rd,7th,10th,and 14th days of administration,and the pictures were processed by MoticMed6 software and Photoshop software;the early wound healing rates of the rats in each group were calculated.The levels of inflammatory factors tumor necrosis factor-α(TNF-α),interleukin-10(IL-10)in the wound were detected by immunohistochemistry,and reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the expression of transforming growth factorβ1 protein(TGF-β1)mRNA and vascular endothelial growth factor(VEGF)mRNA expression in wounds.Western blot(WB)was used to detect the expression of key proteins of p38MAPK signaling pathway p38MAPK,nuclear factor kappa B(NF-κB),cysteine protease 3(caspase-3).Results At 3,7,10 and 14 d after administration,the wound healing rate of Tβ4+hydrocolloid dressing group was higher than that of burn group and p38MAPK agonist group(P<0.05).Compared with the control group,the levels of TNF-αand IL-10 at each time point in the burn group,Tβ4+hydrocolloid dressing group and p38MAPK agonist group were higher(P<0.05).At 12,24,and 48 h after administration,the levels of TNF-αin the burn group and p38MAPK agonist group were higher than those in the Tβ4+hydrocolloid dressing group(P<0.05),while the IL-10 level was lower than that in the Tβ4+hydrocolloid dressing group(P<0.05).However,there was no significant difference in the levels of TNF-αand IL-10 between the burn group and the p38MAPK agonist group at each time point(P>0.05).Compared with the control group,the expressions of TGF-β1 mRNA and VEGF mRNA were all up-regulated at each time point in the burn group,Tβ4+hydrocolloid dressing group and p38MAPK agonist group(P<0.05).Compared with the burn group,the expression of TGF-β1 mRNA was down-regulated at each time point in the Tβ4+hydrocolloid dressing group(P<0.05),while the expression of VEGF mRNA was up-regulated(P<0.05).There was no significant difference in the expressions of TGF-β1 mRNA and VEGF mRNA at each time point between the burn group and the p38MAPK agonist group(P>0.05).Compared with the control group,the expression levels of p38MAPK,NF-κB and caspase-3 in the burn group,Tβ4+hydrocolloid dressing group and p38MAPK agonist group were all higher(P<0.05);Compared with the burn group,the expression levels of p38MAPK,NF-κB and caspase-3 in the Tβ4+hydrocolloid dressing group were all lower(P<0.05).However,there was no significant difference in the expression of p38MAPK,NF-κB and caspase-3 between the burn group and the p38MAPK agonist group(P>0.05).Conclusion Adrenalineβ4 combined with hydrocolloid dressing helps to promote the healing of deepⅡdegree burn wounds,and the mechanism may be related to the inhibition of p38 mitogen-activated protein kinase signal transduction pathway and the reduction of inflammatory factor expression.
作者
李树松
马滢
吴晓明
赵晓玉
李丽
LI Shu-song;MA Ying;WU Xiao-ming;ZHAO Xiao-yu;LI Li(Department of Medical Cosmetology,the Affiliated Hospital of Chengde Medical College,Hebei Province,Chengde 067000,China;Hemodialysis Room,Department of Nephrology,the Affiliated Hospital of Chengde Medical College,Hebei Province,Chengde 067000,China;Department of Burn and Plastic Surgery,the Affiliated Hospital of Chengde Medical College,Hebei Province,Chengde 067000,China)
出处
《河北医科大学学报》
CAS
2024年第7期832-838,共7页
Journal of Hebei Medical University
基金
河北省医学科学研究课题计划(20220009)。
