为明确L-香芹酮的熏蒸抑菌活性和其作用机制,本研究利用菌丝生长速率法测定了L-香芹酮对灰葡萄孢、核盘菌、齐整小核菌、禾谷镰孢、胶孢炭疽菌5种植物病原菌的熏蒸抑制作用,采用孢子萌发法进一步探究了L-香芹酮对灰葡萄孢的作用,通过扫...为明确L-香芹酮的熏蒸抑菌活性和其作用机制,本研究利用菌丝生长速率法测定了L-香芹酮对灰葡萄孢、核盘菌、齐整小核菌、禾谷镰孢、胶孢炭疽菌5种植物病原菌的熏蒸抑制作用,采用孢子萌发法进一步探究了L-香芹酮对灰葡萄孢的作用,通过扫描电镜和透射电镜观察熏蒸处理后灰葡萄孢菌丝微观形态和亚细胞结构的变化,利用PI染色观察细胞膜的损伤情况,并测定L-香芹酮对灰葡萄孢麦角甾醇合成、细胞膜胞外电导率、蛋白质和核酸含量的影响,开展了L-香芹酮对草莓灰霉病的离体防效试验。结果显示,L-香芹酮对5种植物病原菌菌丝生长有明显的抑制作用,熏蒸72 h EC_(50)在5.29~18.69μL/L之间。对灰葡萄孢孢子萌发的EC_(50)为11.78μL/L;经L-香芹酮熏蒸处理后灰葡萄孢菌丝干瘪,扭曲,细胞器结构紊乱;PI染色结果表明,L-香芹酮熏蒸处理严重影响灰葡萄孢细胞膜的完整性,EC_(50)和EC_(90)剂量处理组麦角甾醇含量分别降低27.3%和43.2%,胞内物质外泄,菌丝液的相对电导率上升,蛋白质和核酸类物质外渗程度高于对照。EC_(90)剂量熏蒸处理对草莓灰霉病治疗和保护防控效果分别为85.43%和90.31%。综上,L-香芹酮可通过破坏灰葡萄孢细胞膜的完整性和通透性来发挥抑菌功效,对草莓灰霉病有较好的防治效果,该结果为新型植物源熏蒸杀菌剂的开发提供了新思路。展开更多
Objectives:Among cutaneous malignancies,melanoma stands out for its particularly aggressive nature,with therapeutic interventions becoming notably limited once the disease progresses.In this research,we investigate th...Objectives:Among cutaneous malignancies,melanoma stands out for its particularly aggressive nature,with therapeutic interventions becoming notably limited once the disease progresses.In this research,we investigate the tumor-suppressing capabilities of water-extracted Korean mistletoe(Viscum album var.coloratum)and its purified lectin component(V.album var.coloratum agglutinin,VCA)using two distinct mouse melanoma models:B16BL6 and B16F10 cell lines.Methods:The impact of water extract and VCA treatments on melanoma cells was assessed through multiple experimental approaches,examining cellular survival rates,programmed cell death pathways,multicaspase activity,and cell cycle distribution patterns.To elucidate the interconnections among various cellular responses,we employed a suite of statistical techniques encompassing correlation studies,principal component analysis(PCA)-based dimensionality reduction,and dendrogram-based clusteringmethodologies.Results:Thewater extract exhibited dosedependent cytotoxicity with IC50 values of 372.3±8.7μg/mL and 202.5±8.4μg/mL for B16BL6 and B16F10 cells,respectively.VCA showed more significant effects,with IC50 values of 0.1992±0.0041 and 0.1981±0.0098μg/mL for B16BL6 and B16F10 cells,respectively.Both agents induced substantial apoptosis with a significant progression from early to late apoptotic stages,reaching up to 59.4%total apoptotic cells for VCA treatment.This was confirmed by strong multicaspase activation,particularly in VCA-treated cells(up to 88.4%caspase-positive cells).The water extract showed modest effects on cell cycle distribution,with increases in G0/G1 phase(74.6%)in B16BL6 cells and S phase(19.2%)in B16F10 cells,while VCA treatment resulted in G2/M phase reduction(10.0%)in B16F10 cells.Correlation analysis revealed strong negative associations between cell viability and caspase activity(r=−0.843 to−0.878),while hierarchical clustering demonstrated distinct response patterns between low and high concentrations of both agents.Effect size analysis confirmed strong treatment impacts on cell viability(d=−5.89 to−6.12)and caspase activation(d=3.45 to 5.23).Conclusion:These findings suggest that Korean mistletoe water extract and its isolated lectin may affect both primary and metastatic melanoma cells through distinct mechanisms,demonstrating particular potency in caspase-dependent apoptosis induction.Our findings establish a robust foundation for developing novel therapeutic interventions derived from natural compounds to combat malignant melanoma with high metastatic potential.展开更多
文摘为明确L-香芹酮的熏蒸抑菌活性和其作用机制,本研究利用菌丝生长速率法测定了L-香芹酮对灰葡萄孢、核盘菌、齐整小核菌、禾谷镰孢、胶孢炭疽菌5种植物病原菌的熏蒸抑制作用,采用孢子萌发法进一步探究了L-香芹酮对灰葡萄孢的作用,通过扫描电镜和透射电镜观察熏蒸处理后灰葡萄孢菌丝微观形态和亚细胞结构的变化,利用PI染色观察细胞膜的损伤情况,并测定L-香芹酮对灰葡萄孢麦角甾醇合成、细胞膜胞外电导率、蛋白质和核酸含量的影响,开展了L-香芹酮对草莓灰霉病的离体防效试验。结果显示,L-香芹酮对5种植物病原菌菌丝生长有明显的抑制作用,熏蒸72 h EC_(50)在5.29~18.69μL/L之间。对灰葡萄孢孢子萌发的EC_(50)为11.78μL/L;经L-香芹酮熏蒸处理后灰葡萄孢菌丝干瘪,扭曲,细胞器结构紊乱;PI染色结果表明,L-香芹酮熏蒸处理严重影响灰葡萄孢细胞膜的完整性,EC_(50)和EC_(90)剂量处理组麦角甾醇含量分别降低27.3%和43.2%,胞内物质外泄,菌丝液的相对电导率上升,蛋白质和核酸类物质外渗程度高于对照。EC_(90)剂量熏蒸处理对草莓灰霉病治疗和保护防控效果分别为85.43%和90.31%。综上,L-香芹酮可通过破坏灰葡萄孢细胞膜的完整性和通透性来发挥抑菌功效,对草莓灰霉病有较好的防治效果,该结果为新型植物源熏蒸杀菌剂的开发提供了新思路。
文摘Objectives:Among cutaneous malignancies,melanoma stands out for its particularly aggressive nature,with therapeutic interventions becoming notably limited once the disease progresses.In this research,we investigate the tumor-suppressing capabilities of water-extracted Korean mistletoe(Viscum album var.coloratum)and its purified lectin component(V.album var.coloratum agglutinin,VCA)using two distinct mouse melanoma models:B16BL6 and B16F10 cell lines.Methods:The impact of water extract and VCA treatments on melanoma cells was assessed through multiple experimental approaches,examining cellular survival rates,programmed cell death pathways,multicaspase activity,and cell cycle distribution patterns.To elucidate the interconnections among various cellular responses,we employed a suite of statistical techniques encompassing correlation studies,principal component analysis(PCA)-based dimensionality reduction,and dendrogram-based clusteringmethodologies.Results:Thewater extract exhibited dosedependent cytotoxicity with IC50 values of 372.3±8.7μg/mL and 202.5±8.4μg/mL for B16BL6 and B16F10 cells,respectively.VCA showed more significant effects,with IC50 values of 0.1992±0.0041 and 0.1981±0.0098μg/mL for B16BL6 and B16F10 cells,respectively.Both agents induced substantial apoptosis with a significant progression from early to late apoptotic stages,reaching up to 59.4%total apoptotic cells for VCA treatment.This was confirmed by strong multicaspase activation,particularly in VCA-treated cells(up to 88.4%caspase-positive cells).The water extract showed modest effects on cell cycle distribution,with increases in G0/G1 phase(74.6%)in B16BL6 cells and S phase(19.2%)in B16F10 cells,while VCA treatment resulted in G2/M phase reduction(10.0%)in B16F10 cells.Correlation analysis revealed strong negative associations between cell viability and caspase activity(r=−0.843 to−0.878),while hierarchical clustering demonstrated distinct response patterns between low and high concentrations of both agents.Effect size analysis confirmed strong treatment impacts on cell viability(d=−5.89 to−6.12)and caspase activation(d=3.45 to 5.23).Conclusion:These findings suggest that Korean mistletoe water extract and its isolated lectin may affect both primary and metastatic melanoma cells through distinct mechanisms,demonstrating particular potency in caspase-dependent apoptosis induction.Our findings establish a robust foundation for developing novel therapeutic interventions derived from natural compounds to combat malignant melanoma with high metastatic potential.
