摘要
目的探讨皮粘散治疗慢性皮肤溃疡的可能作用机制及配伍意义。方法将24只SD大鼠随机分为模型组、皮粘散组、水牛皮组、硼砂组,每组6只。各组大鼠通过冰醋酸涂抹缺损性创面制作慢性皮肤溃疡大鼠模型。造模后皮粘散组、水牛皮组、硼砂组分别予以0.2 g药粉均匀地涂抹在溃疡部位,每天换药1次,连续干预21天;模型组不做处理。分别在给药前以及给药的第7、14、21天记录大鼠皮肤溃疡创面面积,计算创面愈合率,并进行溃疡色泽、溃疡分泌物、溃疡肉芽组织评分;末将给药次日采用HE染色观察皮肤组织全层的病理变化;免疫荧光检测大鼠局部溃疡组织中血小板-内皮细胞黏附分子(CD31)、Ⅲ型胶原蛋白(CollagenⅢ)的表达;RT-qPCR法检测大鼠皮肤溃疡创面组织Wnt/β-catenin信号通路相关基因[Wnt家族蛋白4(Wnt4)、连环蛋白β1(Ctnnb1)、G1/S-特异性周期蛋白-D1(Ccnd1)、神经钙黏素(Cdh2)]mRNA表达水平。结果与本组前一时间点比较,皮粘散组、水牛皮组和硼砂组给药第7、14、21天创面面积、皮肤溃疡主要症状积分均降低,给药第14、21天创面愈合率升高(P<0.05)。与模型组同时间点比较,皮粘散组给药第7、14、21天的创面面积及皮肤溃疡主要症状积分均明显降低、创面愈合率明显升高(P<0.05)。给药第21天,皮粘散组CD31、CollagenⅢ免疫荧光可见荧光信号强,且排列一致性较好;HE染色可见胶原纤维排列更加紧密,炎性细胞浸润显著减少,多量新生毛细血管形成;Wnt4、Ctnnb1、Ccnd1、Cdh2 mRNA表达显著增加(P<0.05)。与皮粘散组同时间点比较,水牛皮组及硼砂组给药第7、14、21天创面面积均增大、创面愈合率降低(P<0.05);硼砂组给药第7、14、21天溃疡色泽积分、溃疡肉芽组织积分均显著升高,给药第14、21天溃疡分泌物积分升高(P<0.05)。HE染色可见水牛皮组及硼砂组较皮粘散组炎性细胞浸润显著增加,且硼砂组较水牛皮组炎性细胞浸润更加严重。硼砂组CD31、CollagenⅢ荧光信号弱于水牛皮组;硼砂组Wnt4、Ctnnb1、Ccnd1、Cdh2 mRNA表达较皮粘散组降低(P<0.05)。结论皮粘散对大鼠慢性皮肤溃疡创面有较好的治疗作用,且水牛皮与硼砂配伍具有协同作用,其作用机制可能与调节Wnt/β-catenin信号通路,从而上调创面组织中CD31、CollagenⅢ的表达有关。
Objective To explore the possible mechanism of Pizhan Powder(皮粘散)in the treatment of chronic skin ulcer and the formula composition significance.Methods Twenty-four SD rats were randomly divided into model group,Pizhan Powder group,buffalo skin group,and borax group,with six rats in each group.The rat model of chronic skin ulcer was made by smear of glacial acetic acid on defective wounds.After modeling,0.2 g of corresponding medicinal powder was given to the wounds in the Pizhan Powder group,the buffalo skin group and the borax group,respectively,while the model group was not treated,and the dressing was changed once a day for 21 consecutive days.Before treatment and on the 7th,14th,and 21st day of medication,the skin ulcer wound area,ulcer color,ulcer secretion and granulation tissue of rats were recorded,and the healing rate was calculated.On the next day of last treatment,hematoxylin and eosin(HE)staining was used to observe the full-thickness pathological changes of skin tissue,and immunofluorescence was used to detect the expression of platelet-endothelial cell adhesion molecule(CD31)and typeⅢcollagen(CollagenⅢ)in local ulcer tissue of rats;the mRNA expression of Wnt/β-catenin signaling pathway related genes in skin ulcer wound tissue of rats,including Wnt family protein 4(Wnt4),cateninβ1(Ctnnb1),G1/S-specific cyclin-D1(cyclin-D1),and neural cadherin(Cdh2)were detected by real-time fluorescence quantitative(RT-qPCR).Results Compared to those measured at the previous timepoint in the same group,the ulcer wound area and the main symptom scores of skin ulcers in the Pizhan Powder group,buffalo skin group and borax group significantly decreased on the 7th,14th and 21st day,and the healing rate significantly increased on the 14th and 21st day(P<0.05).Compared to those in the model group measured at the same timepoint,the ulcer wound area and the main symptom scores of skin ulcers on the 7th,14th and 21st day in the Pizhan Powder group significantly decreased,while the healing rate increases(P<0.05).Immunofluorescence showed strong fluorescence signals of CD31 and CollagenⅢwith good alignment on the 21st day in the Pizhan Powder group;HE staining showed that collagen fibers were more closely arranged,and inflammatory cell infiltration was significantly reduced,with formulation of a large amount of new capillaries;the mRNA expressions of Wnt4,Ctnnb1,Ccnd1 and Cdh2 significantly increased(P<0.05).Compared to those in the Pizhan Powder group measured at the same timepoint,the ulcer area of buffalo skin group and borax group on the 7th,14th,and 21st day significantly increased,and the healing rate significantly decreased(P<0.05);the color score and granulation tissue score of ulcers in the borax group significantly increased on the 7th,14th,and 21st day,and the ulcer secretion score on the 14th and 21st day increased(P<0.05).HE staining showed that the infiltration of inflammatory cells in the buffalo skin group and the borax group significantly increased compared to that in the Pizhan Powder group,and more increases were seen in the borax group.Immunofluorescence showed weak fluorescence signals of CD31 and CollagenⅢin the borax group than the buffalo skin group;and mRNA expressions of Wnt4,Ctnnb1,Ccnd1 and Cdh2 significantly decreased in the borax group than the Pizhan Powder group(P<0.05).Conclusion Pizhan Powder has a good therapeutic effect on chronic skin ulcer wound of rats,and the combination of buffalo skin and borax has a coordination effect.The mechanism of action may be related to the up-regulation of the expression of CD31 and CollagenⅢin wound tissues by regulating the Wnt/β-catenin signaling pathway.
作者
宋玮
陈倩倩
唐宋琪
王楷
陈明岭
江英培
肖敏
SONG Wei;CHEN Qianqian;TANG Songqi;WANG Kai;CHEN Mingling;JIANG Yingpei;XIAO Min(Chengdu University of Traditional Chinese Medicine,Chengdu,6610075;Hainan Medical University)
出处
《中医杂志》
CSCD
北大核心
2023年第6期616-624,共9页
Journal of Traditional Chinese Medicine
基金
国家自然科学基金(82004380)
四川省科技厅应用基础研究项目(2019YJ0613)
成都中医药大学“杏林学者”学科人才科研提升计划(QJRC2021007)。
作者简介
通讯作者:肖敏,luckyamy2005@163.com。
