摘要
目的探讨1,25(OH)_(2)D_(3)参与皮肤全层切割伤小鼠创面愈合的作用及机制。方法将40只5周龄C57BL/6雄性小鼠随机分为5组,包括皮肤创伤组、皮肤创伤+维生素D受体(VDR)抑制剂组、皮肤创伤+维生素D受体(VDR)激动剂组、皮肤创伤+维生素D药粉(VD)组、正常对照组,每组8只。观察创面愈合情况,拍照并记录创面大小,计算创面愈合率。在创面处理第9天时,ELISA检测各组小鼠血清中1,25(OH)_(2)D_(3)的浓度;取创缘组织,HE染色检测创面组织病理变化,免疫荧光染色检测组织中VDR、CD31的表达情况,分别用qRT-PCR、Western Blot检测小鼠组织中VDR、Notch1、TNF-α、IL-17A、IL-17F和IL-22的mRNA和蛋白的表达。结果在小鼠伤口愈合第9天时,与正常对照组比较,皮肤创伤组血清1,25(OH)_(2)D_(3)浓度、创面组织VDR、CD31表达降低(P<0.05),TNF-α、IL-17A、IL-17F和IL-22表达升高,Notch1表达差异无统计学意义(P>0.05);与皮肤创伤组比较,皮肤创伤+VDR激动剂组、皮肤创伤+VD组小鼠皮肤创面愈合率、血清1,25(OH)_(2)D_(3)浓度、创面组织VDR、CD31、Notch1表达升高(P<0.05),创面组织TNF-α、IL-17A、IL-17F、和IL-22表达降低(P<0.05),皮肤创伤+VDR抑制剂组小鼠皮肤创面组织TNF-α、IL-22的蛋白表达升高(均P<0.01),创面愈合率、血清1,25(OH)_(2)D_(3)浓度、创面组织VDR、CD31、Notch1、TNF-α、IL-17A、IL-17F、IL-22的mRNA和Notch1、IL-17A、IL-17F的蛋白表达差异无统计学意义(P>0.05)。HE染色结果显示,皮肤创伤+VDR激动剂组、皮肤创伤+VD组小鼠的伤口组织炎性细胞浸润减轻且血管生成增加、真皮结缔组织较致密,表皮基本完成再上皮化。结论1,25(OH)_(2)D_(3)能够促进皮肤全层切割伤小鼠创面愈合,可能是通过减轻创面炎症因子TNF-α、IL-17A、IL-17F和IL-22的表达,促进Notch1受体和VDR的表达和血管生成实现的。
Objective To study the role of 1,25(OH)_(2)D_(3)in cutaneous wound healing and the underlying mechanisms in mice.Methods Forty 5-week-old male C57BL/6 mice were randomly divided into 5 groups,i.e.,wound,wound+vitamin D receptor(VDR)inhibitor,wound+vitamin D receptor(VDR)agonist,wound+vitamin D(VD)powder and normal controls.The wound healing rates were calculated according the photographs of wound size taken at different time points.At the 9th day of wound,ELISA was used to measure serum levels of 1,25(OH)_(2)D_(3).In addition,wound edge tissue was taken for HE staining and immunohistochemical staining of VDR and CD31.Finally,expression levels of both mRNA and proteins for VDR,Notch1,TNF-α,IL-17A,IL-17F and IL-22 were assessed using real-time quantitative PCR and Western Blot,respectively.Results At the 9th day of wound,the levels of serum 1,25(OH)_(2)D_(3)and cutaneous VDR and CD31 expression were decreased,while expression levels of TNF-α,IL-17A,IL-17F and IL-22 were increased in the wounded skin compared to the normal controls(P<0.05).Expression levels of Notch1 did not differ significantly between the normal and wounded skin(P>0.05).Compared with the untreated wounded skin,treatments with either VDR agonist or VD powder significantly increased wound healing rate,serum 1,25(OH)_(2)D_(3)levels and expression levels of cutaneous VDR,CD31 and Notch1,while decreasing cutaneous TNF-α,IL-17A,IL-17F and IL-22 expression(P<0.05).VDR inhibitor increased expression levels of cutaneous TNF-αand IL-22 protein(P<0.01 vs.untreated wound),without significant changes in wound healing rate,serum 1,25(OH)_(2)D_(3)levels,mRNA for cutaneous VDR,CD31,Notch1,TNF-α,IL-17A,IL-17F and IL-22,and protein levels of Notch1,IL-17A and IL-17F.Moreover,the results of HE staining showed that treatments with either VDR agonist or VD powder significantly decreased inflammatory infiltration in the wounded tissue,while increasing angiogenesis along with dense connective tissue in the dermis and almost complete re-epithelialization in the epid
作者
母兴语
何小解
黄诒洁
杭倩如
翁于格
丁艳
MU Xingyu;HE Xiaojie;HUANG Yijie;HANG Qianru;WENG Yuge;DING Yan(First Clinical College of Hainan Medical University,Haikou 571199,China;Children′s Medical Center,Second Xiangya Hospital,Central South University,Changsha 410011,China;Clinical College of Hainan Medical University,Haikou 571199,China;Affiliated Dermatology Hospital of Hainan Medical University,Fifth People′s Hospital of Hainan Province,Haikou 570102,China)
出处
《皮肤性病诊疗学杂志》
2024年第3期147-156,共10页
Journal of Diagnosis and Therapy on Dermato-venereology
基金
海南省重点研发计划项目(ZDYF2020147)。
