基于PI3K/AKT信号通路探究红景天-鬼箭羽药对对人类风湿关节炎成纤维样滑膜细胞的作用机制及配伍规律
Analysis of mechanism of action and compatibility rules of Rhodiola rosea-Eupatorium alatus herb pair in human rheumatoid arthritis fibroblast-like synovial cells through PI3K/AKT signaling pathway
郑秋含 1杜连云 1张众 1朴松兰 2王颖航 3潘志1
作者信息
- 1. 长春中医药大学人参科学研究院,吉林长春 130117
- 2. 长春中医药大学临床医学院,吉林长春 130117
- 3. 长春中医药大学第一附属医院,吉林长春 130117
- 折叠
摘要
目的:基于高效液相色谱法和体外实验探究红景天-鬼箭羽药对(下称"天羽"药对)抗类风湿关节炎(rheumatoid arthritis,RA)的配伍规律及作用机制.方法:通过高效液相色谱法测定"天羽"药对的单煎液以及不同配伍比例的合煎液(2∶1,1∶1,1∶2)中红景天苷的含量.通过体外实验CCK-8、PCR和Western Blot法检测肿瘤坏死因子α诱导的人类风湿关节炎成纤维样滑膜细胞(human fibroblast-like synoviocyte rheumatoid arthritis,HFLS-RA)增殖、mRNA及蛋白表达.结果:"天羽"药对不同配伍比例(2∶1,1∶1,1∶2)的合煎液中红景天苷的含量均高于红景天单煎液,且配伍比例为1∶1时红景天苷的含量最高.CCK-8实验结果表明,"天羽"药对可对HFLS-RA细胞的增殖起到抑制作用.PCR和Western blot实验结果表明,"天羽"药对可显著降低HFLS-RA细胞中p-PI3K、p-AKT、MMP1、MMP3和MMP9 mRNA和蛋白的表达,增加TIMP1 mRNA和蛋白表达.结论:"天羽"药对可有效抑制HFLS-RA细胞增殖,其最佳的配伍比例为1∶1,作用机制可能与抑制PI3K/AKT信号通路,进而调节MMPs/TIMPs平衡有关.
Abstract
OBJECTIVE To explore the efficacy,compatibility rules and mechanism of Rhodiola rosea-Euonymus alatus drug pair("Tianyu"drug pair)for rheumatoid arthritis(RA)through high performance liquid chromatography(HPLC)and in vitro assays.METHODS The contents of salidroside in single decoction of"Tianyu"drug pair and combined decoction of differ-ent compatibility ratios(2∶1,1∶1,1∶2)were determined by HPLC.The proliferation and mRNA/protein expression of TNF-α-induced human fibroblast-like synoviocyte rheumatoid arthritis(HFLS-RA)were detected by CCK-8,polymerase chain reaction(PCR)and Western blot(WB).RESULTS The content of salidroside in combined decoction of"Tianyu"drug pair with differ-ent proportions(2∶1,1∶1,1∶2)was higher than that in single decoction of R.rosea.And the content of salidroside peaked at a compatibility ratio of 1∶1.The results of CCK-8 assay showed that"Tianyu"drug pair could suppress the proliferation of HFLS-RA cells.PCR and WB indicated that the protein/mRNA expressions of p-PI3K,p-AKT,MMP1,MMP3 and MMP9 declined markedly and the mRNA/protein expression of TIMP1 was up-regulated in HFLS-RA cells.CONCLUSION"Tianyu"drug pair can effectively suppress the proliferation of HFLS-RA cells and the optimal compatibility ratio is 1∶1.The mechanism may be related to an inhibition of PI3K/AKT signaling pathway,thereby regulating the balance of MMPs/TIMPs.
关键词
"天羽"药对/人类风湿关节炎成纤维样滑膜细胞/高效液相色谱法/配伍规律/作用机制Key words
"Tianyu"drug pair/human fibroblast-like synoviocyte rheumatoid arthritis/high performance liquid chromatogra-phy/mechanism of action/compatibility rules引用本文复制引用
基金项目
国家自然科学基金(82074324)
吉林省自然科学基金(YDZJ202201ZYTS185)
吉林省自然科学基金(20210101235JC)
吉林省自然科学基金(20230508065RC)
出版年
2024
NETL
NSTL
万方数据