A quantitative analysis method of multi-components with a single marker (QAMS) for simultaneous determination of six marker compounds (one from phenolic acids and five from phthalides) in Chuanxiong Rhizoma was established by applying HPLC and using butylidenephthalide as the internal reference substance. And also the feasibility and accuracy of the established method for quality evaluation and application of Chuanxiong Rhizoma were investigated and validated. The analysis was performed with the mobile phase consisting of acetonitrile - 0.2% aqueous formic acid. The flow rate was 1.0 mL . min-1 and the column temperature was maintained at 30 °C. The detection wavelengths were set at 252 nm (for ferulic acid, Z-ligustilide, and butylidenephthalide) and 266 nm (for senkyunolide I, senkyunolide A, and coniferyl ferulate), separately, and 20 µL was injected for analysis with gradient elution. The results showed that there were no significant differences observed between the HPLC-QAMS method and the external standard method (RSD <5%). The relative correction factors were credible (RSD < 5%) in changed chromatographic conditions. The established HPLC-QAMS method can be accurately used for simultaneously evaluating and controlling the quality of Chuanxiong Rhizoma with multi-components.

Objective: To investigate the protective effect of glucagon-like peptid-1 (GLP-1) against cardiac microvascular endothelial cell (CMECs) injured by high glucose. Methods: CMECs were isolated and cultured. Superoxide assay kit and dihydroethidine (DHE) staining were used to assess oxidative stress. TUNEL staining and caspase 3 expression were used to assess the apoptosis of CMECs. H89 was used to inhibit cAMP/PKA pathway; fasudil was used to inhibit Rho/ROCK pathway. The protein expressions of Rho, ROCK were examined by Western blot analysis. Results: High glucose increased the production of ROS, the activity of NADPH, the apoptosis rate and the expression level of Rho/ROCK in CMECs, while GLP-1 decreased high glucose-induced ROS production, the NADPH activity and the apoptosis rate and the expression level of Rho/ROCK in CMECs, the difference were statistically significant (. P<0.05). Conclusions: GLP-1 could protect the cardiac microvessels against oxidative stress and apoptosis. The protective effects of GLP-1 are dependent on downstream inhibition of Rho through a cAMP/PKA-dependent manner, resulting in a subsequent decrease in the expression of NADPH oxidase.

[摘 要] 目的：探讨谷胱甘肽过氧化物酶（GPX）家族在胶质瘤发生发展中的生物学功能和预后价值。方法：利用TCGA、GTEx和CGGA等多个数据库数据分析胶质瘤中GPX各亚型基因的表达及其相关性、蛋白之间的相互作用、基因突变、GPX表达与胶质瘤患者预后的关系以及GPX7、8的基因集富集分析；GPX8与胶质瘤中免疫细胞浸润以及免疫检查点分子表达的相关性分析，胶质瘤中GPX8表达与化疗药物IC50的相关性分析。采用qPCR法、WB法和免疫荧光技术检测国人胶质瘤组织和对照组织（标本收集自2022年10月20日至12月20日间在上海奉贤区中心医院神经外科手术切除的5例胶质瘤和3例严重脑外伤的病灶组织）中GPX mRNA、蛋白以及相关免疫检查点分子的表达进行验证。结果：数据库分析显示胶质瘤中GPX各亚型蛋白之间存在相互作用、基因表达水平存在相关性（P<0.05或P<0.01），胶质瘤组织中多个GPX亚型存在单核苷酸变异和拷贝数变异；不同类型胶质瘤组织的免疫细胞和肿瘤细胞中主要表达的GPX亚型有明显不同，在胶质瘤组织中GPX1、4、7、8均呈高表达（均P<0.05）且与胶质瘤患者预后不良相关（P<0.01）。qPCR法、WB法检测中国人胶质瘤组织中GPX7、8均呈高表达验证了数据库信息的正确性。在胶质瘤中GPX7、8表达具有独立预后预测价值；富集分析显示GPX7、8与胶质瘤细胞周期和免疫途径有关，在GBM和LGG中GPX8表达与免疫评分呈明显相关（P<0.01）、GPX8可能在胶质瘤中诱导抑制性免疫细胞浸润导致免疫抑制、GPX8表达与胶质瘤中多个免疫检查点分子表达正相关（均P<0.01）、GPX8表达与化疗药物IC50呈明显正相关（均P<0.01）且其高表达可导致胶质瘤对化疗药物的耐药。结论：GPX8在胶质瘤中呈显著高表达，GPX8高表达能诱导胶质瘤中抑制性免疫细胞的浸润其与多个免疫抑制点、与多个化疗药物IC50和患者预后密切相关，可作为胶质瘤免疫治疗的潜在靶点。