| 生物活性 |
Bromuconazole is a triazole fungicide with oral efficacy and blood-brain barrier permeability. Bromuconazole protects crops from various fungal contaminations. Bromuconazole exhibits cytotoxicity against a variety of cancer cells, induces G0/G1 cell cycle arrest and inhibits DNA synthesis in cancer cells, and triggers cytoskeletal structural disorder, genotoxic damage, apoptotic (apoptosis) cell death, and mitochondrial membrane depolarization. Bromuconazole activates caspase-3, induces excessive production of ROS, p53 and Bax, lipid peroxidation, increased activities of SOD and CAT, and downregulates Bcl-2. By upregulating p-ERK1/2 and p-JNK, Bromuconazole disrupts the MAPK signaling pathway, impairs the cellular stress response of human trophoblast cells and endometrial cells, and damages the implantation process. Bromuconazole is applicable to research related to glioma, colon cancer, reproductive injury (implantation dysfunction), and cardiac dysfunction[1][2][3][4][5][6][7].
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体外研究
(In Vitro) |
Bromuconazole (0-500 μM; 24 h) 可 F98、HCT116 细胞中诱导浓度依赖性细胞毒性，IC50 分别为 60、180 μM[1][2]。
Bromuconazole (0-180 μM; 6-24 h) 可 F98、HCT116 细胞中诱导 G0/G1 期细胞周期阻滞并抑制 DNA 合成，诱导细胞骨架结构紊乱、遗传毒性损伤 (DNA 片段化和细胞核固缩)、凋亡性细胞死亡、线粒体膜去极化[1]。
Bromuconazole (15-60 μM; 24 h) 可在大鼠胶质瘤 F98 细胞中诱导 p53 和 Bax mRNA 呈浓度依赖性上调、Bcl-2 mRNA 下调，并使 Bax/Bcl-2 比值升高，诱导 caspase-3 呈浓度依赖性激活[1]。
Bromuconazole (45-180 μM; 24 h) 诱导 HCT116 细胞中 caspase-3 的浓度依赖性激活、ROS 过量生成、脂质过氧化、SOD 和 CAT 的活性升高[2]。
Bromuconazole (0-50 mg/L; 48 h) 会损伤 HTR-8/SVneo 和 T HESCs 的细胞活力，暴露 48 h 后其 LC50 值分别为 28.05 mg/L 和 33.41 mg/L[4]。
Bromuconazole (30 mg/L; 24-48 h) 可破坏 HTR-8/SVneo 和 T HESCs 的 3D 球体形成与自组装，通过调控凋亡相关基因表达异常并升高 BAX/BCL-XL 蛋白比值，诱导细胞发生凋亡，通过下调细胞周期相关基因的表达，诱导 G2/M 期细胞周期阻滞，通过降低线粒体膜电位、下调线粒体功能相关基因的表达，破坏线粒体功能[4]。
Bromuconazole (30 mg/L; 30 min to 24 h) 可通过下调 SOD1 并上调内质网应激相关因子，在 HTR-8/SVneo 和 THESCs 细胞中诱导活性氧 (ROS) 积累与内质网 (ER) 应激[4]。
Bromuconazole (30 mg/L; 15 min) 会通过上调 HTR-8/SVneo、T HESCs 细胞中的 p-ERK1/2 和 p-JNK，来扰乱 MAPK 信号通路[4]。
Bromuconazole (30 mg/L; 24 h) 可通过失调促炎和抗炎细胞因子 mRNA 的表达，在 HTR-8/SVneo 和 THESCs 细胞中诱导炎症反应[4]。
Bromuconazole (30 mg/L; 24-48 h) 可抑制 HTR-8/SVneo 和 T HESCs 的迁移[4]。
Bromuconazole (10-250 μM; 24 h) 可诱导大鼠 H9C2 心肌细胞产生细胞毒性、破坏能量代谢、失调离子平衡及肌球蛋白合成相关基因的表达，并上调 LEF1 蛋白表达[5]。
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
Cell Viability Assay[1]
| Cell Line: |
rat glioma F98 cells |
| Concentration: |
0, 10, 20, 40, 60, 80, 100, 120, 140, 150 μM |
| Incubation Time: |
24 h |
| Result: |
Caused concentration-dependent F98 cell death, with an IC50 of 60 μM after 24 h of treatment. |
Cell Cycle Analysis[1]
| Cell Line: |
rat glioma F98 cells |
| Concentration: |
15, 30, and
60 μM |
| Incubation Time: |
24 h |
| Result: |
Caused dose-dependent accumulation of F98 cells in the G0/G1 phase: at 60 μM, G0/G1 phase cell distribution increased from 64.78% (untreated) to 89%.
Corresponding dose-dependent reduction in S phase cell distribution was observed: at 60 μM, S phase cell distribution decreased from 17.52% (untreated) to 1.5%. |
Apoptosis Analysis[1]
| Cell Line: |
rat glioma F98 cells |
| Concentration: |
15, 30, and
60 μM |
| Incubation Time: |
24 h |
| Result: |
Induced concentration-dependent apoptosis: apoptosis levels were 23.61% at 15 μM, 33.86% at 30 μM, and 55.94% at 60 μM, compared to 1.55% in untreated cells. |
Real Time qPCR[1]
| Cell Line: |
rat glioma F98 cells |
| Concentration: |
15, 30, and
60 μM |
| Incubation Time: |
24 h |
| Result: |
Caused concentration-dependent increases in p53 and Bax mRNA expression, and concentration-dependent decreases in Bcl-2 mRNA expression.
At 60 μM, p53 mRNA levels increased to 2.49-fold, Bax mRNA levels increased to 2.3-fold, Bcl-2 mRNA levels decreased to 0.3-fold, and the Bax/Bcl-2 ratio increased to 7.67-fold. |
Cell Cytotoxicity Assay[2]
| Cell Line: |
human colon carcinoma HCT116 cells |
| Concentration: |
0, 50, 100, 150, 200, 250, 300, 350, 400 μM |
| Incubation Time: |
24 h |
| Result: |
Caused a concentration-dependent increase in HCT116 cell mortality, with an IC50 of 180 μM. |
Cell Cycle Analysis[2]
| Cell Line: |
human colon carcinoma HCT116 cells |
| Concentration: |
45, 90 and 180 μM |
| Incubation Time: |
24 h |
| Result: |
Caused a concentration-dependent accumulation of cells in the G0/G1 phase (from 59.89% in untreated cells to 67.84%, 71.02%, and 84% at 45, 90, and 180 μM, respectively).
Caused a concentration-dependent reduction in cells in the S phase (from 18.9% in untreated cells to 11.03%, 9.26%, and 5.65% at 45, 90, and 180 μM, respectively). |
Apoptosis Analysis[2]
| Cell Line: |
human colon carcinoma HCT116 cells |
| Concentration: |
45, 90 and 180 μM |
| Incubation Time: |
24 h |
| Result: |
Caused a concentration-dependent increase in apoptosis ratio (from 11.15% in untreated cells to 22.45%, 30.92%, and 39.75% at 45, 90, and 180 μM, respectively).
Induced light green staining and crescent-shaped nuclei in treated cells, consistent with apoptotic morphology.\nCaused a concentration-dependent increase in total apoptosis (early + late apoptosis) from 12.29% in untreated cells to 24.5%, 34.02%, and 43.85% at 45, 90, and 180 μM, respectively. |
Cell Viability Assay[4]
| Cell Line: |
HTR-8/SVneo, T HESCs |
| Concentration: |
0, 5, 10, 15, 20, 30, 40, and 50 mg/L |
| Incubation Time: |
48 h |
| Result: |
Reduced cell viability in both cell lines.
Reached an LC50 value of 28.05 mg/L for HTR-8/SVneo cells.
Reached an LC50 value of 33.41 mg/L for T HESCs. |
Apoptosis Analysis[4]
| Cell Line: |
HTR-8/SVneo, T HESCs |
| Concentration: |
0, 10, 20,
30 mg/L (annexin V/PI staining); 30 mg/L (mRNA and protein analysis) |
| Incubation Time: |
48 h (annexin V/PI staining); 24 h (mRNA and protein analysis) |
| Result: |
Increased HTR-8/SVneo early apoptotic cells by 171.58% and late apoptotic cells by 146.96%.
Upregulated HTR-8/SVneo mRNA expression of BAK and CASP1.
Downregulated HTR-8/SVneo mRNA expression of BCL2 and BCL2L1.
Increased the HTR-8/SVneo BAX/BCL-XL protein ratio by 208.17%.
Increased T HESCs early apoptotic cells by 171.58% and late apoptotic cells by 176.12%.
Upregulated T HESCs mRNA expression of BAX and CASP1.
Downregulated T HESCs mRNA expression of BCL2.
Increased the T HESCs BAX/BCL-XL protein ratio by 152.46%. |
Cell Cycle Analysis[4]
| Cell Line: |
HTR-8/SVneo, T HESCs |
| Concentration: |
0, 10, 20, and
30 mg/L (cell cycle analysis); 30 mg/L (mRNA analysis) |
| Incubation Time: |
48 h (cell cycle analysis); 24 h (mRNA analysis) |
| Result: |
Reduced HTR-8/SVneo G0/G1 phase cells to 66.01%.
Increased HTR-8/SVneo S phase cells to 139.83%.
Increased HTR-8/SVneo G2/M phase cells to 118.42%.
Increased HTR-8/SVneo sub G1 phase cells to 234.14%.
Downregulated HTR-8/SVneo mRNA expression of CCNA2, CCNB1, CCND1, CCNE2, CDK1, and CDK4.
Reduced T HESCs G0/G1 phase cells to 85.95%.
Increased T HESCs G2/M phase cells to 138.99%.
Increased T HESCs sub G1 phase cells to 269.69%.
Downregulated T HESCs mRNA expression of CCNA2, CCNB1, CCND1, CCNE2, and CDK1 (no significant change in CDK4). |
Real Time qPCR[4]
| Cell Line: |
HTR-8/SVneo, T HESCs |
| Concentration: |
30 mg/L |
| Incubation Time: |
24 h |
| Result: |
Upregulated HTR-8/SVneo mRNA expression of proinflammatory cytokines IL21, IL21R, and CXCR2.
Downregulated HTR-8/SVneo mRNA expression of anti-inflammatory cytokines IL4R and CCL2.
Upregulated T HESCs mRNA expression of proinflammatory cytokines IL1β, IL18R1, and CXCR2.
Downregulated T HESCs mRNA expression of anti-inflammatory cytokines IL4R and CCL2. |
Cell Migration Assay[4]
| Cell Line: |
HTR-8/SVneo, T HESCs |
| Concentration: |
30 mg/L |
| Incubation Time: |
3-48 h (wound healing assay); 24 h (transwell assay) |
| Result: |
Delayed HTR-8/SVneo wound healing, with 15.78% of initial wound area remaining at 48 h (vs.
1.43% in control).
Reduced HTR-8/SVneo transwell-migrated cells to 52.50%.
Delayed T HESCs wound healing, with 7.61% of initial wound area remaining at 48 h (vs.
1.84% in control). |
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体内研究
(In Vivo) |
Bromuconazole (13.8-32.8 mg/kg/天；口服；每日；90 天；84-200 mg/kg/天；局部给药；每日；90 天) 可在雄性白化大鼠中诱导剂量依赖性肝毒性，且口服途径的毒性强于局部给药；其特征包括肝酶水平升高、肝脏氧化应激、肝脏重量增加、组织病理学上的坏死性病变、肝脏 PXR 与 CYP3A1 表达及 CYP3A1 活性上调，以及肝脏 CAR 与 CYP2B1 表达下调[3]。
Bromuconazole (50 ng/L-7.5 mg/L；i.g.；每日一次；连续 7 天 ) 暴露会通过扰乱心脏能量代谢、改变离子平衡与肌球蛋白合成基因表达，以及异常激活 LEF1，导致成年雄性 AB 品系斑马鱼产生心脏毒性[5]。
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
| Animal Model: |
albino rats (male, adult, 160-200 g)[3] |
| Dosage: |
13.8 mg/kg/day (oral); 32.8 mg/kg/day (oral); 84 mg/kg/day (topical); 200 mg/kg/day (topical) |
| Administration: |
p.o.; daily; 90 days; topical; daily; 90 days |
| Result: |
Increased serum ALT, AST , ALP , ACP, total bilirubin, direct bilirubin, indirect bilirubin, and liver MDA.
Decreased serum total protein, albumin, and liver SOD.
Increased liver weigh.
Induced 5.3-fold increase in hepatic CYP3A1 mRNA expression, 3.4-fold increase in PXR mRNA expression, 4.3-fold decrease in CYP2B1 mRNA expression, 2.7-fold decrease in CAR mRNA expression, and ~7-fold increase in hepatic CYP3A1 enzyme activity.
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| Animal Model: |
AB-strain (adult male, 4-6 months old, weight 450 mg, body length 38 mm)[5] |
| Dosage: |
50 ng/L; 7.5 mg/L |
| Administration: |
i.g.; daily; 7 days |
| Result: |
Increased fatness index significantly.
Reduced body length.
Caused looser heart structure, increased ventricular and arteriole volume, thinner heart walls, more vacuoles in the ventricular cavity, and increased numbers of hypertrophic cardiomyocytes with reduced cardiomyocyte density in both dose groups.
Identified 684 differentially expressed genes, with significant enrichment in cardiac energy metabolism pathways.
Increased heart tissue triglycerides; reduced ATP content in a concentration-dependent manner.
Slightly increased TG, reduced TC and low-density lipoprotein (LDL), and increased high-density lipoprotein (HDL).
Reduced mRNA expression of glycolysis genes GK and HK1, lipid transport gene abca1b, and UCP2.
Downregulated ion balance-related genes ryr2b, atp2a2a, and Pln in a concentration-dependent manner.
Upregulated myosin synthesis-related gene myl4, while downregulated myh7l, tpm4b, and tnnt2a.
Activated LEF1 gene expression abnormally in heart tissue compared to controls.
Increased cardiac ejection fraction significantly compared to controls. |
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