PDF(Pubmed)
Abstract：
BACKGROUND: The leaves of Kalanchoe pinnata (Lam.) Pers. (K. pinnata), a succulent plant native to tropical regions, are used as a medicinal alternative against cancer in several countries worldwide; however, its therapeutic potential to fight cancer has been little addressed. In this study, we analyzed the phytochemical content, antioxidant capacity, and selectivity of K. pinnata leaf ethanolic extract against different human cancer cell lines in vitro.
METHODS: This study subjected the ethanolic extract to enzymatic assays to quantify the phytochemical content (phenolics, flavonoids, and anthraquinones) and its radical scavenging and iron-reducing capacities. Also, the phytoconstituents and major phenolic compounds present in the extract\'s subfractions were identified by GC-MS, HPLC, and NMR. Human cancer (MCF-7, PC-3, HT-29) and normal colon (CoN) cell lines were treated with different concentrations of K. pinnata leaf ethanolic extract, and the changes in cell proliferation (sulforhodamine B assay), caspases activity (FITC-VAD-FMK reporter), mitochondrial membrane potential (MMP, rhodamine 123 assay), chromatin condensation/fragmentation (Hoechst 33342 stain), and ROS generation (DCFH2 probe assay) were assessed.
RESULTS: The results showed that the K. pinnata leaf ethanolic extract is rich in phytoconstituents with therapeutic potential, including phenols (quercetin and kaempferol), flavonoids, fatty acid esters (34.6% of the total composition), 1- triacontanol and sterols (ergosterol and stigmasterol, 15.4% of the total composition); however, it presents a poor content of antioxidant molecules (IC50 = 27.6 mg/mL for H2O2 scavenging activity vs. 2.86 mg/mL in the case of Trolox). Notably, the extract inhibited cell proliferation and reduced MMP in all human cell lines tested but showed selectivity for HT-29 colon cancer cells compared to CoN normal cells (SI = 8.4). Furthermore, ROS generation, caspase activity, and chromatin condensation/fragmentation were augmented significantly in cancer-derived cell lines, indicating a selective cytotoxic effect.
CONCLUSIONS: These findings reveal that the K. pinnata leaf ethanolic extract contains several bioactive molecules with therapeutic potential, capable of displaying selective cytotoxicity in different human cancer cell lines.
METHODS: This study subjected the ethanolic extract to enzymatic assays to quantify the phytochemical content (phenolics, flavonoids, and anthraquinones) and its radical scavenging and iron-reducing capacities. Also, the phytoconstituents and major phenolic compounds present in the extract\'s subfractions were identified by GC-MS, HPLC, and NMR. Human cancer (MCF-7, PC-3, HT-29) and normal colon (CoN) cell lines were treated with different concentrations of K. pinnata leaf ethanolic extract, and the changes in cell proliferation (sulforhodamine B assay), caspases activity (FITC-VAD-FMK reporter), mitochondrial membrane potential (MMP, rhodamine 123 assay), chromatin condensation/fragmentation (Hoechst 33342 stain), and ROS generation (DCFH2 probe assay) were assessed.
RESULTS: The results showed that the K. pinnata leaf ethanolic extract is rich in phytoconstituents with therapeutic potential, including phenols (quercetin and kaempferol), flavonoids, fatty acid esters (34.6% of the total composition), 1- triacontanol and sterols (ergosterol and stigmasterol, 15.4% of the total composition); however, it presents a poor content of antioxidant molecules (IC50 = 27.6 mg/mL for H2O2 scavenging activity vs. 2.86 mg/mL in the case of Trolox). Notably, the extract inhibited cell proliferation and reduced MMP in all human cell lines tested but showed selectivity for HT-29 colon cancer cells compared to CoN normal cells (SI = 8.4). Furthermore, ROS generation, caspase activity, and chromatin condensation/fragmentation were augmented significantly in cancer-derived cell lines, indicating a selective cytotoxic effect.
CONCLUSIONS: These findings reveal that the K. pinnata leaf ethanolic extract contains several bioactive molecules with therapeutic potential, capable of displaying selective cytotoxicity in different human cancer cell lines.
摘要：
背景：Kalanchoepinnata的叶子（Lam。)Pers。(K.pinnata),一种原产于热带地区的多汁植物，在世界几个国家被用作抗癌的药物替代品；然而，它对抗癌症的治疗潜力几乎没有得到解决。在这项研究中,我们分析了植物化学物质的含量，抗氧化能力，和选择性的K.pinnata叶乙醇提取物对不同的人癌细胞系的体外。
方法：本研究对乙醇提取物进行酶测定，以定量植物化学物质含量（酚类，黄酮类化合物，和蒽醌）及其自由基清除和铁还原能力。此外，通过GC-MS鉴定了提取物亚组分中存在的植物成分和主要酚类化合物，HPLC,和NMR。用不同浓度的K.pinnata叶乙醇提取物处理人类癌症（MCF-7，PC-3，HT-29）和正常结肠（CoN）细胞系，和细胞增殖的变化（磺罗丹明B测定），胱天蛋白酶活性（FITC-VAD-FMK报告基因），线粒体膜电位(MMP，罗丹明123测定)，染色质凝聚/碎片化（Hoechst33342染色），和ROS产生(DCFH2探针测定)被评估。
结果：结果表明，山葵叶乙醇提取物富含具有治疗潜力的植物成分，包括酚类（槲皮素和山奈酚），黄酮类化合物，脂肪酸酯（占总组成的34.6％），1-三十烷醇和甾醇（麦角甾醇和豆甾醇，占总成分的15.4%)；然而，它呈现的抗氧化剂分子含量低（IC50=27.6mg/mL的H2O2清除活性与在Trolox的情况下为2.86mg/mL）。值得注意的是,提取物在所有测试的人细胞系中抑制细胞增殖并降低MMP,但与CoN正常细胞相比显示对HT-29结肠癌细胞的选择性(SI=8.4)。此外，ROS生成，caspase活性，在癌症来源的细胞系中，染色质凝聚/片段显着增强，表明选择性细胞毒性作用。
结论：这些发现表明，山葵叶乙醇提取物含有几种具有治疗潜力的生物活性分子，能够在不同的人类癌细胞系中显示选择性细胞毒性。
方法：本研究对乙醇提取物进行酶测定，以定量植物化学物质含量（酚类，黄酮类化合物，和蒽醌）及其自由基清除和铁还原能力。此外，通过GC-MS鉴定了提取物亚组分中存在的植物成分和主要酚类化合物，HPLC,和NMR。用不同浓度的K.pinnata叶乙醇提取物处理人类癌症（MCF-7，PC-3，HT-29）和正常结肠（CoN）细胞系，和细胞增殖的变化（磺罗丹明B测定），胱天蛋白酶活性（FITC-VAD-FMK报告基因），线粒体膜电位(MMP，罗丹明123测定)，染色质凝聚/碎片化（Hoechst33342染色），和ROS产生(DCFH2探针测定)被评估。
结果：结果表明，山葵叶乙醇提取物富含具有治疗潜力的植物成分，包括酚类（槲皮素和山奈酚），黄酮类化合物，脂肪酸酯（占总组成的34.6％），1-三十烷醇和甾醇（麦角甾醇和豆甾醇，占总成分的15.4%)；然而，它呈现的抗氧化剂分子含量低（IC50=27.6mg/mL的H2O2清除活性与在Trolox的情况下为2.86mg/mL）。值得注意的是,提取物在所有测试的人细胞系中抑制细胞增殖并降低MMP,但与CoN正常细胞相比显示对HT-29结肠癌细胞的选择性(SI=8.4)。此外，ROS生成，caspase活性，在癌症来源的细胞系中，染色质凝聚/片段显着增强，表明选择性细胞毒性作用。
结论：这些发现表明，山葵叶乙醇提取物含有几种具有治疗潜力的生物活性分子，能够在不同的人类癌细胞系中显示选择性细胞毒性。
