Kikowska Małgorzata, Piotrowska-Kempisty Hanna, Kucińska Małgorzata, Murias Marek, Budzianowski Jaromir, Budzianowska Anna, Kaczmarek Mariusz, Kowalczyk Mariusz, Stochmal Anna, Thiem Barbara
Laboratory of Pharmaceutical Biology and Biotechnology, Department and Division of Practical Cosmetology and Skin Diseases Prophylaxis, Poznan University of Medical Sciences, Collegium Pharmaceuticum, 3 Rokietnicka St., 60-806 Poznan, Poland.
Department of Toxicology, Poznan University of Medical Sciences, Dojazd 30, 60-631 Poznań, Poland.
Plants (Basel). 2023 Jun 29;12(13):2485. doi: 10.3390/plants12132485.
(1) The cytotoxicity and antioxidant activity of different fractions as well as the pro-apoptotic activity of saponin fractions from L. in SKOV-3 was investigated. (2) In screening studies, the cytotoxicity of six fractions on SKOV-3 was examined by LDH and SRB assays. The most active fractions-triterpenoid saponins-were selected for further investigation. To determine the mechanism of saponin fractions' cytotoxicity, their ability to induce apoptosis was examined via Annexin V assay. The effect of the saponin fractions on caspase 3 activity was measured using a Caspase 3 Assay Kit. The expression of 84 apoptosis-related genes was investigated in cancer cells exposed to saponin fractions from the roots. The radical scavenging capacity of different fractions was determined via DPPH assay. (3) The pronounced cytotoxic effects in SKOV-3 were demonstrated by saponin fractions from the leaves and roots. Those saponin fractions were chosen for further investigation. The treatment of cancer cell lines with saponins obtained from the roots provoked a significant increase in apoptotic cells. In the SKOV-3 cells, saponins caused upregulation of pro-apoptotic genes and a decrease in anti-apoptotic genes. The activation of caspase 3 was correlated with an increased DFFA expression level in the treated SKOV-3 cells. The most active fractions were phenolic acids from the shoots and roots. (4) To the best of our knowledge, the current study is the first to demonstrate that the barrigenol-type triterpenoid saponin fraction from the roots of inhibits SKOV-3 cell proliferation and induces apoptosis, which may be regulated by the expression of genes mostly specific to a mitochondria-related pathway.
(1) 研究了不同部位提取物的细胞毒性、抗氧化活性以及地锦草皂苷部位在SKOV-3细胞中的促凋亡活性。(2) 在筛选研究中，通过乳酸脱氢酶（LDH）和磺酰罗丹明B（SRB）测定法检测了六个部位提取物对SKOV-3细胞的细胞毒性。选择活性最强的部位——三萜皂苷——进行进一步研究。为了确定皂苷部位细胞毒性的机制，通过膜联蛋白V测定法检测其诱导凋亡的能力。使用半胱天冬酶3检测试剂盒测定皂苷部位对半胱天冬酶3活性的影响。研究了暴露于根部皂苷部位的癌细胞中84个凋亡相关基因的表达。通过二苯基苦味酰基自由基（DPPH）测定法确定不同部位提取物的自由基清除能力。(3) 叶和根部位的皂苷提取物在SKOV-3细胞中表现出显著的细胞毒性作用。选择这些皂苷部位进行进一步研究。用根部获得的皂苷处理癌细胞系导致凋亡细胞显著增加。在SKOV-3细胞中，皂苷导致促凋亡基因上调和抗凋亡基因下调。半胱天冬酶3的激活与处理后的SKOV-3细胞中DNA片段化因子α（DFFA）表达水平的增加相关。活性最强的部位是茎和根中的酚酸。(4) 据我们所知，本研究首次证明地锦草根中的巴里金醇型三萜皂苷部位可抑制SKOV-3细胞增殖并诱导凋亡，这可能受主要与线粒体相关途径特异的基因表达调控。
