文献类型：期刊文献

中文题名：敦煌芮草膏对Ⅱ度冻伤大鼠的治疗作用及抗炎作用机制

英文题名：Preliminary Study on the Therapeutic Effect and Anti-inflammatory Mechanism of Dunhuang Ruicao Plaster on Rats with Second Degree Frostbite

作者：张莉[1];尚芸[1];王强[1];杜籼芹[1];李爽[1];李亮亮[1];高榕妮[1];蔺兴遥[1,2]

第一作者：张莉

机构：[1]甘肃中医药大学基础医学院,兰州730000;[2]敦煌医学与转化教育部重点实验室,兰州730000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2024

卷号：44

期号：6

起止页码：709

中文期刊名：中国中西医结合杂志

外文期刊名：Chinese Journal of Integrated Traditional and Western Medicine

收录：CSTPCD;;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；PubMed;

基金：敦煌医学开发转化人才培养与膏摩方芮草膏的开发研究(No.2021RCXM106);敦煌医学膏摩方芮草膏的开发研究(No.DHYX19-14)。

语种：中文

中文关键词：敦煌芮草膏;冻伤;药效;机制;中药

外文关键词：Dunhuang Ruicao Plaster;frostbite;efficacy;mechanism;Chinese herbal medicine

摘要：目的探究敦煌芮草膏对冻伤的治疗作用及机制。方法去将96只SPF级SD大鼠用随机数字表法分为6组,每组16只。除空白组之外,剩余80只大鼠采用液氮冷冻法造模,分为5组,即模型(M)组,敦煌芮草膏低(DRP-L)中(DRP-M)高剂量(DRP-H)组,磺胺嘧啶银乳膏(SSC)组。除空白组外,其余组每日擦涂给药。于造模后第3、7天取材,每组8只。检测冻伤大鼠皮损大体情况、创口愈合率、肿胀率、病理检查;ELISA法检测大鼠血清中促炎因子肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-1β、血管内皮生长因子(VEGF)表达;采用实时荧光定量PCR、WesternBlot法检测局部创面组织中Toll样受体4(TLR4),髓样分化因子88(MyD88),核转录因子κBp65(NF-κBp65)mRNA及蛋白表达。结果果与空白组比较,第3、7天,M组大鼠血清中IL-1β、IL-6、TNF-α的含量及VEGF表达量均明显升高(P<0.05),创面组织中TLR4、MyD88、NF-κBp65 mRNA及蛋白表达增高(P<0.05),M组大鼠炎症反应明显。与M组比较,在造模第3天,DRP-L组NF-κB mRNA水平及MyD88蛋白水平降低(P<0.05);DRP-M、DRP-H、SSC组TNF-α、IL-6含量及TLR4、MyD88mRNA和蛋白水平(P<0.05,P<0.01)及NF-κB、p-NF-κB蛋白水平降低(P<0.05,P<0.01);且SSC组可IL-1β含量降低(P<0.05),VEGF水平升高(P<0.05),NF-κB mRNA水平降低(P<0.05)。造模第7天,DRP-L组可TNF-α、IL-6含量降低(P<0.05),VEGF水平升高(P<0.01),NF-κB mRNA表达水平降低(P<0.05),MyD88、p-NF-κB蛋白水平降低(P<0.01);DRP-M、H组TNF-α、IL-1β、IL-6水平(P<0.05,P<0.01),升高VEGF水平(P<0.01);降低TLR4、MyD88、NF-κB mRNA及蛋白水平降低(P<0.05,P<0.01);SSC组IL-1β降低(P<0.05),VEGF水平升高(P<0.01),NF-κB mRNA水平降低,MyD88、NF-κB、p-NF-κB蛋白水平降低(P<0.05,P<0.01)。结论敦煌芮草膏可能通过抑制TLR4/MyD88/NF-κB信号通路的激活,减少炎症因子的释放和表达,来发挥冻伤的治疗作用。
Objective e To study the therapeutic effect and mechanism of Dunhuang Ruicao Plaster(DRP)on frostbite.Methods The 96 SPF SD rats were divided into 6 groups,16 in each group.The remaining 80 rats(except those in the blank group)were modeled using liquid nitrogen freeze,and further divided into five groups,i.e.,the model group(M),low dose DRP group(DRP-L),medium dose DRP group(DRP-M),high dose DRP group(DRP-H),sulfonate silver cream(SSC)group.Except those in the blank group,mice in the rest groups were rubbed daily.The rats were sampled on the 3rd and 7th day after modeling,8 rats in each group.The general condition of skin lesion,wound healing rate,sweling rate,pathological examinations,and the expressions of tumor necrosis factor(TNF)-αa,IL-6,IL-1β,and vascular endothelial growth factor(VEGF)in serum of rats were detected by enzyme-linked immunosorbent assay(ELiISA).mRNA and protein expressions of Toll-like receptors 4(TLR4),myeloid differentiation primary response gene 88(MyD88),nuclear factor kappa-B(NF-κB)p65 in local wound tissue were detected by Real-time fluorescence quantitative PCR and Western Blot respectively.Results Compared with the blank group,the serum contents of IL-1β,IL-6,and TNF-αand the expression of VEGF significantly increased(P<0.05),and mRNA and protein expressions of TLR4,MyD88,NF-κB p65 in the wound tissue significantly increased in the M group on the 3rd and 7th day(P<0.05).The inflammatory response in the M group was obvious.Compared with the M group,the expression of NF-κB mRNA and MyD88 protein significantly decreased in the DRP-L group on the 3rd day of modeling(P<0.05).The contents of TNF-αand IL-6,mRNA and protein expression levels of TLR4 and MyD88,protein expression levels of NF-κB and p-NF-κB were reduced in DRP-M,DRP-H,and SSC groups(P<0.05,P<0.01).IL-1βcontent decreased(P<0.05)and VEGF level increased(P<0.05),and NF-κB mRNA level decreased(P<0.05)in SSC group.On the 7th day of modeling,the levels of TNF-αand IL-6 decreased(P<0.05),the level of VEGF increased(P<0.01),the level of NF-κB mRNA reduced,and the levels of MyD88 and p-NF-κB protein lowered in DRP-L group(P<0.01).The levels of TNF-αa,IL-1β,and IL-6 decreased(P<0.05,P<0.01),the level of VEGF increased(P<0.01),mRNA and protein levels of TLR4,MyD88,and NF-κB decreased(P<0.05,P<0.01)in DRP-M and DRP-H groups.IL-1βsignificantly decreased(P<0.05),VEGF level increased(P<0.01),NF-κB mRNA level reduced,and protein levels of MyD88,NF-κB,and p-NF-κB lowered(P<0.05,P<0.01)in SSC group.Conclusion DRP played a role in the treatment of frostbite by inhibiting the activation of TLR4/MyD88/NF-κB signaling pathway,and attenuating the release and expression of inflammatory factors.