风热清口服液对免疫低下小鼠免疫调节作用的研究

目的:探究风热清口服液对免疫低下小鼠免疫调节的作用。方法:通过给小鼠腹腔注射免疫抑制剂环磷酰胺50 mg/kg,1次/d,连续3 d,建立免疫功能低下的动物模型。应用碳廓清实验、溶血素生成实验和迟发型变态反应,评价风热清口服液对免疫低下小鼠非特异性免疫、体液免疫和细胞免疫的调节作用。结果:风热清口服液低、中剂量组可不同程度提高环磷酰胺致免疫低下小鼠的廓清指数K值(P<0.05)和吞噬指数α值(P<0.05)。风热清口服液低、中、高剂量组可不同程度提高免疫低下小鼠血清溶血素水平(P<0.05)。风热清口服液低、中剂量组可不同程度促进免疫低下小鼠迟发型变态反应(P<0.05)。风热清口服液低剂量组可提高免疫低下小鼠脾脏中CD4^(+)细胞数(P<0.01),并使CD4^(+)/CD8^(+)细胞比值上升(P<0.001),同时抑制免疫低下小鼠脾脏中CD8^(+)细胞数(P<0.05);风热清口服液中、高剂量组可提高免疫低下小鼠脾脏中CD4^(+)/CD8^(+)细胞比值(P<0.01),同时抑制免疫低下小鼠脾脏中CD8^(+)细胞数(P<0.05)。风热清口服液低、中剂量组可显著提高免疫低下小鼠脾指数(P<0.001),风热清口服液低、中、高剂量组均可提高免疫低下小鼠胸腺指数(P<0.05或P<0.01)。结论:风热清口服液通过提高免疫低下小鼠非特异性免疫、体液免疫和细胞免疫,进而增强免疫低下小鼠的免疫功能。

基于网络药理学研究三棱-莪术药对抗子宫内膜癌的作用机制及实验验证

目的:利用网络药理学、分子对接技术,探讨三棱-莪术药对抗子宫内膜癌的作用机制,并通过人子宫内膜癌细胞ISK进行体外试验验证重要信号通路。方法:在TCMSP平台筛选出三棱-莪术药对主要活性成分;通过TCMSP数据库和Swiss Target Prediction得到活性成分的靶点;在GeneCards等数据库检索疾病的作用靶点;利用Cytoscape3.7.2软件建立可视化药物-活性成分-关键靶点调控网络及PPI网络;用Metascape数据库和OmicShare tools对关键靶点进行GO富集与KEGG分析;并通过PDB数据库与Discovery Studio 2016软件对活性成分和核心靶点进行分子对接。最后通过ISK细胞对重要信号通路进行验证。结果:从三棱-莪术药对筛选出活性成分11个,包括去氢木香内酯、芒柄花黄素、豆甾醇、反式甘多酸、谷固醇、秦皮甲素、β-榄香烯、双去甲氧基姜黄素、莪术醇、异莪术烯醇、温金素,药物与疾病共同的关键靶点92个,核心靶点6个,包括RAC-α丝氨酸/苏氨酸蛋白激酶、原癌基因酪氨酸蛋白激酶Src、丝裂原激活的蛋白激酶1、雌激素受体、热休克蛋白HSP 90-alpha、丝裂原激活的蛋白激酶8。GO富集与KEGG分析三棱-莪术药对活性成分主要参与癌症的途径、癌症中的蛋白聚糖、PI3K-AKT信号通路等。分子对接提示芒柄花黄素、双去甲氧基姜黄素、温金素三个活性成分与核心靶蛋白结合能力较好。CCK8结果表明,与空白对照组比较,双去甲氧基姜黄素能显著抑制ISK细胞的增殖(P<0.01);在显微镜下可观察到,双去甲氧基姜黄素3、6、9μg/mL组ISK细胞数量明显减少,Western blot结果显示,与空白对照组比较,双去甲氧基姜黄素9μg/mL组能明显下调PI3K、AKT蛋白的表达(P<0.05)。结论:三棱-莪术药对可能主要通过芒柄花黄素、双去甲氧基姜黄素、温金素调节细胞代谢、细胞增殖与凋亡等途径发挥抗子宫内膜癌作用。

Anti-bacterial and anti-viral effects of Fengreqing oral liquid(风热清口服液) in vitro and in vivo

OBJECTIVE:To investigate the anti-bacterial and anti-viral effects of Fengreqing oral liquid(风热清口服液,FRQ)in vitro and in vivo.METHODS:The minimum inhibitory concentrations of Fengreqing Oral Liquid against six gram-positive bacteria(Staphylococcus aureus,Streptococcus mutans,Peptostreptococcus anaerobius,Hemolytic streptococcus,Streptococcus pneumoniae,Klebsiella pneumoniae),seven gram-negative bacteria(Escherichia coli,Actinobacillus actinomycetemcomitans,Porphyromonas gingivalis,Haemophilus influenzae,Helicobacter pylori,Pseudomonas aeruginosa,Gardnerella vaginalis)and Candida albicans were detected by the paper disc diffusion method.The inhibition rate of A/Puerto Rico/8/34(H1N1)(PR8)influenza virus in different concentrations of Fengreqing oral solution was detected by chicken embryo method.CCK8 method was used to detect the half-cell infection of RSV,VSV and CVB3.The effect of FRQ on the survival curve of mice was detected by using co-infection model of Streptococcus pneumoniae and influenza virus.RESULTS:In vitro,FRQ can inhibit Actinobacillus actinomycetemcomitans,Helicobacter pylori,Gardnerella vaginalis,Staphylococcus aureus,Streptococcus mutans and Streptococcus pneumoniae and has an antiviral effect on the envelope virus H1N1.In vivo,Fengreqing oral solution had therapeutic effect on influenza-Streptococcus pneumoniae co-infection in mice,significantly improving the survival rate of mice.The medium dose and low dose FRQ significantly prolonged the survival time of mice.CONCLUSION:FRQ has good anti-bacterial and anti-viral effects in vivo and in vitro.