摘要
目的研究八味中药的体外抑菌效果,筛选出外用治疗痤疮的最佳复方配比,制备复方微乳凝胶用于小鼠痤疮模型的体内药效学研究。方法选取痤疮丙酸杆菌和金黄色葡萄球菌为目标菌,采用滤纸片法测定八味中药的抑菌圈直径和最小抑菌浓度(MIC);采用正交试验法优化复方配比;以伪三元相图为评价指标制备微乳,对微乳及微乳凝胶理化性质进行综合评价;采用背部注射痤疮丙酸杆菌建立小鼠痤疮模型,设置空白组、模型组、阳性药组、微乳凝胶组和复方微乳凝胶组,通过HE和Masson染色观察组织病理改变,酶联免疫吸附试验(ELISA)检测血清中炎症因子白介素1β(IL-1β)、白介素10(IL-10)和肿瘤坏死因子-α(TNF-α)水平。结果结果表明八味中药均有不同程度的抑菌效果,其中黄连对痤疮丙酸杆菌的抑菌效果最好,甘草对金黄色葡萄球菌的抑菌效果最好,MIC均为31.25 mg/mL。正交试验结果表明,大黄、黄连、黄芩、甘草以250、250、125、62.5 mg/mL,即配比为4∶4∶2∶1时杀菌率最佳。根据伪三元相图得到最终微乳处方为吐温-803 g,聚乙二醇4006 g,肉豆蔻酸异丙酯1 g,蒸馏水12 g,按质量比加入1%卡波姆940,充分溶胀后得到微乳凝胶,将水相用中药复方提取液代替得到复方微乳凝胶。药效学实验表明,与空白组相比,模型组小鼠皮肤组织出现明显的病理改变,血清中IL-1β、IL-10、TNF-α含量显著增高(P<0.01);与模型组相比,给药组病理性改变有不同程度的缓解,血清炎症因子含量降低,其中复方微乳凝胶组改善效果最显著(P<0.01)。结论以大黄、黄连、黄芩、甘草配伍的复方具有明显的抑菌作用,制备复方微乳凝胶对小鼠痤疮模型有良好的治疗作用,可以有效缓解痤疮丙酸杆菌引发的痤疮炎症反应。
Objective To study the antibacterial effect of eight kinds of Chinese drugs in vitro,screen the best compound ratio for external use to treat acne and prepare compound microemulsion gel for in vivo pharmacodynamic study of acne model in mice.Methods Propionibacterium acnes and Staphylococcus aureus were selected as target bacteria,and the diameter of inhibition zone and minimum inhibitory concentration(MIC)of eight Chinese drugs were determined by filter paper method.Orthogonal test was used to optimize the compound ratio.The physical and chemical properties of microemulsion and microemulsion gel were compre-hensively evaluated by preparing microemulsion with pseudo ternary phase diagram as evaluation index.A mouse acne model was established by injecting Propionibacterium acnes into the back,and the blank group,model group,positive drug group,microemul-sion gel group and compound microemulsion gel group were set up.Histopathological changes were observed by HE and Masson staining,and the levels of inflammatory factors[interleukin-1β(IL-1β),interleukin-10(IL-10)and tumor necrosis factor-(TNF-α)]in serum were detected by enzyme-linked immunosorbent assay(ELISA).Results The results showed that all the eight Chinese drugs had different antibacterial effects,among which Huanglian(Coptidis Rhizoma)had the best antibacterial effect on Propionibacterium acnes and Gancao(Glycyrrhizae Radix et Rhizoma)had the best antibacterial effect on Staphylococ-cus aureus,with MIC of 31.25 mg mL-1.The orthogonal test results showed that the sterilization rate of Dahuang(Rhei Radix et 85第43卷第3期2025年3月中华中医药学刊CHINESE ARCHIVES OF TRADITIONAL CHINESE MEDICINE Vol.43 No.3 Mar.2025 Rhizoma)(250 mg mL-1),Huanglian(Coptidis Rhizoma)(250 mg mL-1),Huangqin(Scutellariae Radix)(125 mg mL-1)and Gancao(Glycyrrhizae Radix et Rhizoma)(62.5 mg mL-1)was the best when the ratio was 4∶4∶2∶1.According to the pseudo-ternary phase diagram,the final microemulsion formula was Tween-80(3 g),PEG 400(6 g),isopropyl myristate(1 g)and distilled water(12 g),and 1%carbomer 940 was added according to the mass ratio,and the microemulsion gel was obtained after full swelling,and the compound microemulsion gel was obtained by replacing the water phase with the compound extract of the Chinese medicine compound.The pharmacodynamic experiment showed that compared with those of the blank group,the skin tissue of the model group showed obvious pathological changes,and the contents of IL-1β,IL-10 and TNF-αin serum in-creased significantly(P<0.01).Compared with those in the model group,the pathological changes in the drug group were re-lieved to varying degrees,and the contents of serum inflammatory factors were decreased,especially those in the compound micro-emulsion gel group(P<0.01).Conclusion The compound of Dahuang(Rhei Radix et Rhizoma),Huanglian(Coptidis Rhizo-ma),Huangqin(Scutellariae Radix)and Gancao(Glycyrrhizae Radix et Rhizoma)has obvious bacteriostatic effect.The prepara-tion of the compound microemulsion gel has a good therapeutic effect on acne model in mice,and can effectively alleviate acne in-flammation caused by Propionibacterium acnes.
作者
梁进京
杨新雪
董蕊
LIANG Jinjing;YANG Xinxue;DONG Rui(College of Chinese Materia Medica,Jilin Agricultural University,Changchun 130118,Jilin,China)
出处
《中华中医药学刊》
北大核心
2025年第3期85-91,I0014-I0019,共13页
Chinese Archives of Traditional Chinese Medicine
基金
国家中药标准化项目(ZYBZH-Y-JL-25)。
关键词
痤疮
加味三黄泻心汤
正交试验
抑菌实验
微乳凝胶
药效学
acne
Modified Sanhuang Xiexin Decoction(加味三黄泻心汤)
orthogonal test
bacteriostasis experiment
micro-emulsion gel
pharmacodynamics
