Darzi Salimeh Ebrahimnezhad, Khazraei Seyedeh Parvin, Amirghofran Zahra
Department of Immunology, Shiraz University of Medical Sciences, Shiraz, I.R. Iran.
Medicinal and Natural Products Chemistry Research Center and Autoimmune Disease Research Center, Shiraz University of Medical Sciences, Shiraz, I.R. Iran.
Res Pharm Sci. 2018 Apr;13(2):103-110. doi: 10.4103/1735-5362.223792.
( Apiaceae) is an important plant currently used in folk medicine to treat various diseases including infections and inflammatory conditions. In this study we have investigated the immunomodulatory and apoptosis-inducing activities of the fractions from this plant. We prepared dichloromethane, hexane, butanol, and water fractions from the aerial parts of the plant. We examined the growth inhibitory effects of the fractions on proliferative lymphocytes by the BrdU incorporation assay. Cell viability was determined by the propidium iodide (PI) assay. AnnexinV/PI staining and cell cycle analysis by flow cytometry was performed to investigate the apoptosis-inducing effects of the plant fractions. Cytokine levels were measured by enzyme-linked immunosorbent assay. All fractions reduced cell proliferation of the activated lymphocytes. The dichloromethane fraction with the highest inhibitory effect (IC, 19.8 μg/mL) significantly reduced cell viability. Although the butanol fraction inhibited cell growth (IC, 88.2 μg/mL) it did not affect cell viability. Annexin V/PI and cell cycle analysis showed the maximum apoptosis-inducing effect of the dichloromethane fraction at 200 μg/mL. Cytokines, that included interleukin (IL)-4 (22.8 ± 0.9 pg/mL, < 0.05) and interferon (IFN)-γ (651 ± 37.5 pg/mL, < 0.01) reduced the activated lymphocyte levels in cultures in the presence of 100 μg/mL concentration of butanol fraction compared to the untreated control. In conclusion, the dichloromethane fraction of had the capability of inducing apoptosis in the activated lymphocytes, whereas the butanol fraction reduced cell activation and cytokine secretion. These data suggested the potential of these examined fractions for more studies in terms of their beneficial effects on immune-mediated diseases.
伞形科植物是目前民间医学中用于治疗包括感染和炎症性疾病在内的各种疾病的重要植物。在本研究中，我们调查了该植物各馏分的免疫调节和诱导凋亡活性。我们从该植物地上部分制备了二氯甲烷、己烷、丁醇和水馏分。我们通过BrdU掺入试验检测了各馏分对增殖淋巴细胞的生长抑制作用。通过碘化丙啶（PI）试验测定细胞活力。进行膜联蛋白V/PI染色和流式细胞术细胞周期分析以研究植物馏分的诱导凋亡作用。通过酶联免疫吸附测定法测量细胞因子水平。所有馏分均降低了活化淋巴细胞的细胞增殖。抑制作用最强的二氯甲烷馏分（IC，19.8μg/mL）显著降低了细胞活力。虽然丁醇馏分抑制细胞生长（IC，88.2μg/mL），但它不影响细胞活力。膜联蛋白V/PI和细胞周期分析显示，二氯甲烷馏分在200μg/mL时诱导凋亡的作用最强。与未处理的对照相比，在存在100μg/mL浓度丁醇馏分的培养物中，包括白细胞介素（IL）-4（22.8±0.9pg/mL，<0.05）和干扰素（IFN）-γ（651±37.5pg/mL，<0.01）在内的细胞因子降低了活化淋巴细胞水平。总之，该植物的二氯甲烷馏分具有诱导活化淋巴细胞凋亡的能力，而丁醇馏分降低了细胞活化和细胞因子分泌。这些数据表明，这些检测馏分在对免疫介导疾病的有益作用方面具有进一步研究的潜力。
