寒喘祖帕单煎配方与配伍合煎对16HBE细胞抗炎活性及非靶向代谢组学差异的研究  

Study on difference ofanti-inflammatory activity and non-targeted metabolomics of 16HBE cells between a single decoction formulation of Hanchuan Zupa and its combination

作　　者：曹万萍 李新霞[1,2] 李乔乔 马雪红 闫冬[1,4] CAO Wanping;LI Xinxia;LI Qiaoqiao;MA Xuehong;YAN Dong(College of Pharmacy,Xinjiang Medical University,Urumqi 830000,China;Xinjiang Key Laboratory of Active Components and Drug Release Technology of Natural Medicines,Urumqi 830011,China;Pharmacy Department,Xi′an High-Tech Hospital,Xi′an 710075,China;Laboratory and Equipment Management Department,Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆医科大学药学院 [2]新疆天然药物活性组分与释药技术重点实验室,乌鲁木齐830017 [3]西安高新医院药剂科,西安710075 [4]新疆医科大学实验室与设备管理处,乌鲁木齐830017

出　　处：《新疆医科大学学报》2024年第4期575-583,共9页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区科技计划项目(2021A03002-2);中央引导地方科技发展专项项目(ZYYD2023B11)。

摘　　要：目的考察寒喘祖帕按照传统配伍合煎及单煎配方的抗炎活性,并采用UPLC-Q-TOF/MS研究非靶向代谢组学差异。方法通过体外脂多糖诱导16HBE炎症细胞,使用CCK-8法筛选出单煎配方组和配伍合煎组的剂量;采用逆转录酶聚合酶链反应(Reverse Transcription-Polymerase Chain Reaction,RT-PCR)技术测定细胞中炎症因子IL-6、IL-8和TNF-α的mRNA表达水平,并采用UPLC-TOF/MS技术对细胞进行非靶向代谢组学分析:分为正常组(16HBE细胞+培养基)、模型组(16HBE细胞+培养基+0.1μg/mL LPS)、寒喘祖帕单煎配方组(16HBE细胞+培养基+0.1μg/mL LPS+50μg/mL单煎配方)、寒喘祖帕配伍合煎组(16HBE细胞+培养基+0.1μg/mL LPS+50μg/mL配伍合煎)。以此探究寒喘祖帕颗粒发挥抗炎活性的相关代谢通路,寻找寒喘祖帕颗粒抗炎活性相关的差异代谢物。结果与正常组比较,模型组在给药浓度为0.1μg/mL时细胞生长存活率未受到限制,差异无统计学意义(P>0.05);配伍合煎组在给药浓度为0~400μg/mL范围内时,细胞存活率未受影响,浓度>800μg/mL时,细胞生长存活率下降,差异具有统计学意义(P<0.01);而单煎配方组在浓度<50μg/mL时,细胞生长存活率未受到限制,浓度≥100μg/mL时,细胞生长存活率下降,差异具有统计学意义(P<0.01);RT-PCR实验结果显示:与正常组比较,模型组的IL-6、IL-8和TNF-αmRNA的表达上升,差异具有统计学意义(P<0.001),50μg/mL单煎配方较100μg/mL,IL-6、IL-8和TNF-αmRNA的表达下降,差异具有统计学意义(P<0.001);在相同药物浓度干预下,与正常组比较,模型组的IL-6、IL-8和TNF-αmRNA的表达上升,差异具有统计学意义(P<0.001),与模型组比较,单煎配方组和配伍合煎组可同时降低IL-6、IL-8和TNF-αmRNA的表达,差异具有统计学意义(P<0.001);配伍合煎组与单煎配方组比较,配伍合煎组IL-6、IL-8 mRNA的表达较单煎配方低,差异具有统计学意义(P<0.05)。代谢组学分析结果筛选出模�Objective To examined the anti-inflammatory activity of Hanchuan Zupa according to the traditional combination of decoction and single decoction formulations,and to studied the non-targeted metabolomics differences by UPLC-Q-TOF/MS.Methods 16HBE inflammatory cells were induced by lipopolysaccharide in vitro,and the dosages of single decoction formula group and the combined decoction group were screened using CCK-8 method;the mRNA expression levels of inflammatory factors IL-6,IL-8 and TNF-αin the cells were determined using Reverse(reverse transcription-polymerase chain reaction,RT-PCR)was used to determine the mRNA expression levels of the inflammatory factors IL-6,IL-8 and TNF-αin the cells,and UPLC-TOF/MS was used to analyze the cells for non-targeted metabolomics.They were divided into the normal group(16HBE cells+medium),the model group(16HBE cells+medium)and the model group(16HBE cells+medium).The cells were divided into normal group(16HBE cells+culture medium),model group(16HBE cells+culture medium+0.1μg/mL LPS),Hanchuan Zupa single decoction formula group(16HBE cells+culture medium+0.1μg/mL LPS+50μg/mL single decoction formula),and Hanchuan Zupa combined decoction group(16HBE cells+culture medium+0.1μg/mL LPS+50μg/mL combined decoction).To investigate the metabolic pathways involved in the anti-inflammatory activity of Hanchuan Zupa granules,and to search for the differential metabolites associated with the anti-inflammatory activity of Hanchuan Zupa granules.Results Compared with the normal group,the cell growth and survival rate of the model group was not restricted at a concentration of 0.1μg/mL,and the difference was not statistically significant(P>0.05).In the combined decoction group,the cell growth and survival rate was not affected at a concentration of 0~400μg/mL,and the cell growth and survival rate wasdecreased at a concentration of≥800μg/mL,and the difference was statistically significant(P<0.01);while in the single decoction group,the cell growth and survival rate was not restrict

关 键 词：寒喘颗粒 单煎配方 配伍合煎 炎症 非靶向代谢组学 

分 类 号：R965[医药卫生—药理学]