A strategy of screening and binding analysis of bioactive components from traditional Chinese medicine based on surface plasmon resonance biosensor / 药物分析学报

Elucidating the active components of traditional Chinese medicine(TCM)is essential for understanding the mechanisms of TCM and promote its rational use as well as TCM-derived drug development.Recent studies have shown that surface plasmon resonance(SPR)technology is promising in this field.In the present study,we propose an SPR-based integrated strategy to screen and analyze the major active components of TCM.We used Radix Paeoniae Alba(RPA)as an example to identify the compounds that can account for its anti-inflammatory mechanism via tumor necrosis factor receptor type 1(TNF-R1).First,RPA extraction was analyzed using an SPR-based screening system,and the potential active in-gredients were collected,enriched,and identified as paeoniflorin and paeonol.Next,the affinity con-stants of paeoniflorin and paeonol were determined as 4.9 and 11.8 μM,respectively.Then,SPR-based competition assays and molecular docking were performed to show that the two compounds could compete with tumor necrosis factor-α(TNF-α)while binding to the subdomain 1 site of TNF-R1.Finally,in biological assays,the two compounds suppressed cytotoxicity and apoptosis induced by TNF-α in the L929 cell line.These findings prove that SPR technology is a useful tool for determining the active in-gredients of TCM at the molecular level and can be used in various aspects of drug development.The SPR-based integrated strategy is reliable and feasible in TCM studies and will shed light on the eluci-dation of the pharmacological mechanism of TCM and facilitate its modernization.

miR-17-5p regulates the proliferation and apoptosis of myelodysplastic syndrome SKM-1 cells by targeting SETD2 / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨miR-17-5p和含SET结构域蛋白2（SETD2）对骨髓增生异常综合征（MDS）SKM-1细胞增殖与凋亡的影响及其作用机制。方法：收集2019年3月至2021年5月衡水市人民医院就诊的35例MDS患者的骨髓标本（MDS组）、35例健康体检者的骨髓标本（对照组），以及MDS细胞系SKM-1。用qPCR法检测MDS骨髓和SKM-1细胞中miR-17-5p、SETD2 mRNA的表达水平。双荧光素酶报告基因实验验证miR-17-5p与SETD2的靶向关系。利用脂质体转染技术，分别将si-miR-NC、si-miR-17-5p、miR-NC、miR-17-5p mimic、pcDNA、pcDNA-SETD2、si-miR-17-5p+si-NC、si-miR-17-5p+si-SETD2等转染至SKM-1细胞，CCK-8法、流式细胞术检测细胞的增殖和凋亡水平，WB法检测细胞中SETD2、C-caspase-3、C-caspase-9的表达。结果：与对照组相比，MDS组骨髓中miR-17-5p表达水平显著升高、SETD2的mRNA和蛋白表达水平均显著降低（均P<0.01）。与si-miR-NC组相比，si-miR-17-5p组SKM-1细胞增殖能力显著降低、凋亡率显著升高，细胞中C-caspase-3和C-caspase-9表达显著升高（均P<0.01）。miR-17-5p明显抑制野生型SETD2细胞的荧光素酶活性（P<0.01），并负向调控SETD2的表达。过表达SETD2可显著抑制SKM-1细胞的增殖并促进细胞凋亡，同时干扰SETD2表达则可部分逆转干扰miR-17-5p对SKM-1细胞的增殖抑制和凋亡促进作用。结论：MDS骨髓中miR-17-5p呈高表达，干扰miR-17-5p可抑制SKM-1细胞增殖并促进细胞凋亡，其机制与miR-17-5p靶向负调控SETD2的表达有关。