苦参碱通过Hippo信号通路促进大鼠皮肤创面愈合及血管生成  

Matrine promotes skin wound healing and angiogenesis through Hippo signaling pathway in rats

作　　者：曾华[1] 郑勇[1] 周林[2] 冯上新 唐俊[1] 孙妍婧 罗玮 ZENG Hua;ZHENG Yong;ZHOU Lin;FENG Shangxin;TANG Jun;SUN Yanjing;LUO Wei(Department of Anorectal Medicine,Wuhan Hospital of Traditional Chinese Medicine,Wuhan 430014,Hubei,China;Department of Pathology,Wuhan Hospital of Traditional Chinese Medicine,Wuhan 430014,Hubei,China;Department of General Medicine,The Third People's Hospital of Hubei Province,Wuhan 430030,Hubei,China)

机构地区：[1]武汉市中医医院肛肠科,武汉430014 [2]武汉市中医医院病理科,武汉430014 [3]湖北省第三人民医院全科医学科,武汉430030

出　　处：《医学研究与战创伤救治》2024年第1期23-31,共9页Journal of Medical Research & Combat Trauma Care

基　　金：武汉市卫生健康委科研项目(WZ21Q24)。

摘　　要：目的探讨苦参碱(MT)对皮肤创面愈合的影响及可能机制。方法在48只大鼠中成功建立全层皮肤切除创伤模型,将模型大鼠随机分为阴性对照组(模型组,涂抹100μL等渗盐水)、阳性药物组(PD组,涂抹100μL的京万红软膏)、苦参碱组(MT组,涂抹浓度为200 ng/mL的苦参碱混合液100μL)、苦参碱+维替泊芬组(MT+VP组,涂抹苦参碱混合液100μL后,腹腔注射100 mg/kg维替泊芬),每组12只。各组每日给药1次,持续7 d。比较各组大鼠的创面愈合率、病理组织损伤情况、CD31阳性染色细胞比例,以及血清白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)水平。蛋白免疫印迹检测(Western blot)Hippo通路相关蛋白表达。结果造模后3、7、14 d时,与模型组创面愈合率[(14.02±1.87)%、(36.23±3.62)%、(65.53±3.85)%]相比,PD组[(21.36±2.51)%、(61.86±3.13)%、(94.26±0.26)%]和MT组[(21.57±2.24)%、(54.35±3.66)%、(94.18±3.13)%]均明显升高(P<0.05);与MT组相比,MT+VP组创面愈合率[(18.35±2.03)%、(43.68±3.54)%、(74.26±3.55)%]均明显降低(P<0.05)。造模后7 d,与模型组(186.75±18.53、85.73±10.15、57.96±5.85)相比,PD组(89.75±9.75、43.12±5.75、8.76±1.13)和MT组(92.57±23.62、46.62±6.54、12.36±1.06)的IL-1β、IL-6、TNF-α水平均明显降低(P<0.05),CD31阳性染色细胞百分比明显升高(P<0.05),MST1、LAST1和YAP蛋白磷酸化水平明显升高(P<0.05),而YAP核移位明显降低(P<0.05)。与MT组相比,MT+VP组IL-1β、IL-6、TNF-α水平均明显升高,CD31阳性染色细胞百分比明显降低,MST1、LAST1和YAP蛋白磷酸化水平明显降低,而YAP核移位明显增加(P<0.05)。与模型组皮肤创面完全愈合时间[(26.33±0.94)d]和瘢痕厚度[(0.58±0.11)mm]相比,PD组[(19.50±0.82)d、(0.24±0.04)mm]和MT组[(19.00±0.50)d、(0.25±0.04)mm]明显减少(P<0.05);与MT组相比,MT+VP组创面完全愈合时间[(22.33±0.94)d]和瘢痕厚度[(0.41±0.04)mm]均明显增加(P<0.05)。结论苦参碱Objective To investigate the effect of matrine(MT)on skin wound healing and its possible mechanism.Methods A total skin excision trauma model was successfully constructed in 48 rats.The model rats were randomly divided into a negative control group(Model group,smeared with 100μL isotonic saline),a positive drug group(PD group,smeared with 100μL Jing Wan Hong ointment),a matrine group(MT group,100μL matrine mixed solution with a concentration of 200 ng/mL),and a matrine+verteporfin group(MT+VP group,intraperitoneal injection of 100 mg/kg verteporfin after applying 100μL matrine mixture),with 12 rats in each group.Each group was administered once daily for 7 d.The wound healing rate,pathological tissue damage,proportion of CD31 positively stained cells,and serum IL-6,TNF-α,and IL-1βlevels were compared among the groups.Western blot was performed to detect the expression of Hippo pathway-related proteins.Results On the 3rd,7th,and 14th day after modeling,compared with the wound healing rates of the model group[(14.02±1.87)%,(36.23±3.62)%,(65.53±3.85)%],PD group[(21.36±2.51)%,(61.86±3.13)%,(94.26±0.26)%]and MT group[(21.57±2.24)%,(54.35±3.66)%,(94.18±3.13)%]were significantly increased(P<0.05);Compared with the MT group,the wound healing rate of the MT+VP group[(18.35±2.03)%,(43.68±3.54)%,(74.26±3.55)%]was significantly decreased(P<0.05).7 days after modeling,compared with the model group(186.75±18.53,85.73±10.15,57.96±5.85),the levels of IL-1β,IL-6 and TNF-αin PD group(89.75±9.75,43.12±5.75,8.76±1.13)and MT group(92.57±23.62,46.62±6.54,12.36±1.06)were significantly decreased(P<0.05),the percentage of CD31 positive staining cells,the phosphorylation levels of MST1,LAST1 and YAP proteins were significantly increased(P<0.05),while the nuclear translocation of YAP was significantly decreased(P<0.05).Compared with the MT group,the MT+VP group had significant increases in the levels of IL-1β,IL-6,and TNF-α,a significant reduction in the percentage of CD31 positive staining cells,a significan

关 键 词：皮肤创面愈合 苦参碱 炎症 血管生成 Hippo通路 

分 类 号：R282[医药卫生—中药学]