Zhang Lijuan, Siyiti Mukadaisi, Zhang Jiang, Yao Meiqi, Zhao Feicui
Department of Pharmacy, Xinjiang Medical University, Urumqi, Xinjiang 830011, P.R. China.
Department of Pharmacy, The Affiliated Chinese Medicine Hospital of Xinjiang Medical University, Urumqi, Xinjiang 830000, P.R. China.
Exp Ther Med. 2021 May;21(5):493. doi: 10.3892/etm.2021.9924. Epub 2021 Mar 17.
The aim of the present study was to investigate the cell proliferation-inhibiting and anti-rheumatic activities of chemical components from Chemical constituents of were separated and purified by silica gel and Sephadex LH-20 chromatography. Structure was identified by spectroscopic technique, and physical/chemical properties were analyzed. The following four compounds were identified: i) Aconitine, ii) songorine, iii) 16, 17-dihydro-12β, 16β-epoxynapelline, and iv) 12-epi-napelline. Cell Counting kit-8 assay was performed to assess cell proliferation. ELISA was conducted to determine the cytokine contents, and reverse transcription-quantitative polymerase chain reaction and Western blot analysis were performed to detect the mRNA and protein expression levels. Compared with the lipopolysaccharide (LPS) group, the contents of IL-6, IL-1β, TNF-α and PGE-2 in the culture supernatant were significantly declined in the leflunomide + LPS and intervention+LPS groups, as well as the mRNA expression levels of HIF-1α, VEGFA and TLR4. Treatments with songorine, benzoylaconine and aconitine (at different concentrations) significantly inhibited the proliferation of HFLS-RA cells. Compared with the LPS group, the contents of PGE-2, IL-6, IL-1β and TNF-α in the culture supernatant were significantly decreased in the intervention groups, and the mRNA expression levels of TLR4, HIF-1α and VEGFA in the cells in the intervention groups. Songorine, benzoylaconine and aconitine from have anti-rheumatic activities , which may inhibit the proliferation of HFLS-RA cells, and the underlying mechanisms may be associated with inhibiting the inflammatory cytokine production and downregulating the expression levels of HIF-1α, VEGF and TLR4.
本研究的目的是研究 的化学成分的细胞增殖抑制和抗风湿活性。 的化学成分通过硅胶和葡聚糖凝胶LH - 20柱色谱进行分离和纯化。通过光谱技术鉴定结构，并分析物理/化学性质。鉴定出以下四种化合物：i）乌头碱，ii）松果碱，iii）16,17 - 二氢 - 12β，16β - 环氧那可丁，和iv）12 - 表那可丁。进行细胞计数试剂盒 - 8检测以评估细胞增殖。进行酶联免疫吸附测定以确定细胞因子含量，并进行逆转录 - 定量聚合酶链反应和蛋白质印迹分析以检测mRNA和蛋白质表达水平。与脂多糖（LPS）组相比，来氟米特 + LPS组和干预 + LPS组培养上清液中IL - 6、IL - 1β、TNF - α和PGE - 2的含量以及HIF - 1α、VEGFA和TLR4的mRNA表达水平均显著下降。用松果碱、苯甲酰乌头原碱和乌头碱（不同浓度）处理显著抑制了类风湿关节炎成纤维样滑膜细胞（HFLS - RA细胞）的增殖。与LPS组相比，干预组培养上清液中PGE - 2、IL - 6、IL - 1β和TNF - α的含量以及干预组细胞中TLR4、HIF - 1α和VEGFA的mRNA表达水平均显著降低。 中的松果碱、苯甲酰乌头原碱和乌头碱具有抗风湿活性，可能抑制HFLS - RA细胞的增殖，其潜在机制可能与抑制炎性细胞因子产生和下调HIF - 1α、VEGF和TLR4的表达水平有关。
