中药新药与临床药理2025，Vol.36Issue(6)：907-917,11.DOI:10.19378/j.issn.1003-9783.2025.06.008
苦豆子总碱凝胶调控巨噬细胞极化促进糖尿病大鼠伤口愈合的作用机制研究
Mechanism of Total Alkaloids of Sophora Alopecuroides Gel in Promoting Wound Healing in Diabetic Rats by Regulating Macrophage Polarization
摘要
Abstract
Objective To investigate the mechanism of total alkaloids of Sophora Alopecuroides(TAS)gel in promoting wound healing in diabetic rats.Methods(1)A diabetic rat model was established by intraperitoneal injection of Streptozotocin(STZ,60 mg·kg-1),and a full-thickness skin defect model was created by excising the dorsal skin.Forty rats were randomly divided into a control group,a model group,a low-dose TAS group(1%TAS gel),a high-dose TAS group(2%TAS gel),and a positive control group(recombinant bovine basic fibroblast growth factor gel),with eight rats in each group.The low-and high-dose TAS groups and the positive control group were treated with the corresponding gels,while the control and model groups were treated with blank gel,once daily for 12 days.HE staining was used to observe pathological changes in wound tissues;Masson staining and immunohistochemistry were used to detect collagen deposition and angiogenesis in wound tissues;TUNEL staining was used to detect apoptosis in wound tissues;immunofluorescence staining was used to measure the expressions of F4/80+,CD86+,and CD206+cells in wound tissues;ELISA was used to detect the levels of tumor necrosis factor-α(TNF-α),transforming growth factor-β1(TGF-β1),interleukin(IL)-1β,and IL-10 in wound tissues.(2)THP-1 cells were polarized into M1/M2 phenotypes using lipopolysaccharide(LPS)/interferon-γ(IFN-γ)and IL-4.The cells were divided into an M0 group,an M1 group,an M2 group,a low-dose TAS group(M1/M2+10 µg·mL-1 TAS),a high-dose TAS group(M1/M2+20 µg·mL-1 TAS),and a glycolysis inhibitor(2-DG)group(M1+2 mmol·L-1 2-DG).After 48 hours of treatment,flow cytometry was used to detect the percentages of CD86+and CD206+cells.A lactate assay kit was used to measure lactate levels in M1-polarized THP-1 cells;Western Blot was used to detect the protein expressions of hexokinase 2(HK2)and glucose transporter 1(Glut1).Results(1)Compared with the control group,the model group showed unclosed wound epithelium,disordered tissue structure,massive inflammatory cell infiltration,incomplete neovascularization,reduced collagen content,and decreased neovascularization(P＜0.01).Compared with the model group,the TAS-treated groups showed closed wound epithelium,reduced disordered tissue structure,inflammatory cell infiltration,and incomplete neovascularization,and significantly increased collagen content and neovascularization(P＜0.01).Compared with the control group,the model group showed increased wound non-healing rate,TUNEL-positive cell percentage,and CD86+/(F4/80+)percentage(P＜0.01),elevated TNF-α and IL-1β levels(P＜0.01),decreased CD206+/(F4/80+)percentage(P＜0.01),and reduced IL-10 and TGF-β1 levels(P＜0.05,P＜0.01).Compared with the model group,the TAS-treated groups showed reduced TUNEL-positive cell percentage and CD86+/(F4/80+)percentage(P＜0.01),decreased TNF-α and IL-1β levels(P＜0.05,P＜0.01),increased CD206+/(F4/80+)percentage(P＜0.01),and elevated IL-10 and TGF-β1 levels(P＜0.05,P＜0.01).(2)Compared with the M0 group,the M1 group showed increased CD86+percentage and lactate levels(P＜0.01),upregulated HK2 and Glut1 protein expression(P＜0.01),and the M2 group showed increased CD206+percentage(P＜0.01).Compared with the M2 group,the TAS-treated group showed reduced CD206+percentage(P＜0.01);compared with the M1 group,the TAS-treated and 2-DG groups showed reduced CD86+percentage(P＜0.01),and the TAS-treated group showed decreased lactate levels and HK2 and Glut1 protein expression(P＜0.05).Conclusion TAS gel can inhibit inflammatory responses,promote angiogenesis and collagen deposition in wound tissues,and accelerate diabetic wound healing.Its anti-inflammatory mechanism is related to the inhibition of glycolysis metabolism and regulation of macrophage M1/M2 phenotypic polarization.关键词
糖尿病/皮肤缺损/苦豆子总碱/伤口愈合/巨噬细胞/抗炎/糖酵解/大鼠Key words
diabetes mellitus/skin defect/total alkaloids of Sophora Alopecuroides/wound healing/macrophages/anti-inflammatory/glycolysis/rats分类
医药卫生引用本文复制引用
涂彩凤,古红婷,邝莹,陈垒..苦豆子总碱凝胶调控巨噬细胞极化促进糖尿病大鼠伤口愈合的作用机制研究[J].中药新药与临床药理,2025,36(6):907-917,11.基金项目
国家自然科学基金地区项目(82160777,82260803). （82160777,82260803）
