Beneficial effects of natural compounds are often attributed to modulation of NO production;however effects produced by plant extracts do not correlate with effects of purified components. The goal of our work was to ...Beneficial effects of natural compounds are often attributed to modulation of NO production;however effects produced by plant extracts do not correlate with effects of purified components. The goal of our work was to study ability of flavonoids and indole-3-carbinol, as well as their combinations to modify NO production, iNOS gene and protein expression in rat tissues. Baicalein and luteolin decreased NO concentration in both intact and LPS-treated animals. Baicalein decreased iNOS gene expression. Luteolin decreased NO production in the liver and heart and number of iNOS-positive cells in the liver of LPS-treated animals. Combination of the two substances did not decrease the NO synthesis triggered by LPS, although it decreased iNOS gene expression. Quercetin decreased NO production in the heart, kidneys and blood of intact rats, but enhanced the LPS effect in testes, spleen and blood on NO production and iNOS protein expression in the liver. Indole-3-carbinol decreased NO concentration in the cerebellum, blood, lungs and skeletal muscles. The drug enhanced the LPS-triggered increase of NO production in all rat organs. It increased iNOS protein expression in intact liver;however it decreased the LPS-triggered outburst of the enzyme biosynthesis. Combination of indole3-carbinol with quercetin decreased NO production in LPS-treated animals however it slightly increased iNOS gene expression. Taken together our results suggest that modifications of NO level in tissues by a natural compound cannot be predicted from data about its effects on NOS expression or activity. Combination of substances can produce an effect differing from that of individual substances. This stresses importance of direct measurements of NO in the tissues.展开更多
Indole-3-carbinol(I3C),an important anticancer compound found in broccoli,has attracted considerable attention.The rapid extraction and accurate analysis of I3C in the pharmaceutical industry in broccoli is challengin...Indole-3-carbinol(I3C),an important anticancer compound found in broccoli,has attracted considerable attention.The rapid extraction and accurate analysis of I3C in the pharmaceutical industry in broccoli is challenging as I3C is unstable at low pH and high temperature.In this study,a rapid,accurate,and lowcost ultrasound-assisted dispersive-filter extraction(UADFE)technique based on poly(deep eutectic solvent)-graphene oxide(PDES-GO)adsorbent was developed for the isolation and analysis of I3C in broccoli for the first time.PDES-GO with multiple adsorption interactions and a fast mass transfer rate was synthesized to accelerate adsorption and desorption.UADFE was developed by combining dispersive solid-phase extraction(DSPE)and filter solid-phase extraction(FSPE)to realize rapid extraction and separation.Based on the above two strategies,the proposed PDES-GO-UADFE method coupled with high-performance liquid chromatography(HPLC)allowed the rapid(15-16 min),accurate(84.3%-96.4%),and low-cost(adsorbent:3.00 mg)analysis of I3C in broccoli and was superior to solid-phase extraction,DSPE,and FSPE methods.The proposed method showed remarkable linearity(r=0.9998;range:0.0840-48.0 mg/g),low limit of quantification(0.0840 mg/g),and high precision(relative standard deviation<5.6%).Therefore,the PDES-GO-UADFE-HPLC method shows significant potential in the field of pharmaceutical analysis for the separation and analysis of anti-cancer compounds in complex plant samples.展开更多
【目的】探讨吲哚-3-原醇(indol-3-carbinol,I3C)对人低分化鼻咽癌细胞株CNE-2的作用及其机制。【方法】将对数生长期的CNE-2细胞分成空白对照组和I3C组。空白对照组细胞常规培养,I3C组细胞培养体系中加入I3C(50μmol/L)。各组细胞培养4...【目的】探讨吲哚-3-原醇(indol-3-carbinol,I3C)对人低分化鼻咽癌细胞株CNE-2的作用及其机制。【方法】将对数生长期的CNE-2细胞分成空白对照组和I3C组。空白对照组细胞常规培养,I3C组细胞培养体系中加入I3C(50μmol/L)。各组细胞培养48 h后,采用四甲基偶氮唑盐(MTT)法检测细胞增殖;Annexin V-碘化丙啶/异硫氰酸荧光素(PI/FITC)检测细胞凋亡,Real-time PCR检测细胞中胞外信号调节激酶(ERK)和Bax、Bcl-2基因表达;Western blot法检测细胞中ERK、p-ERK、Bax、Bcl-2蛋白表达,Caspase3活性试剂盒检测Caspase3活性。【结果】I3C组与空白对照组比较,CNE-2细胞增殖率显著下降(P<0.05),凋亡率显著升高(P<0.05),Real-time PCR检测显示I3C组与空白对照组比较,ERK m RNA表达无显著性差异,但Bax m RNA表达水平显著增高(P<0.001),Bcl-2 m RNA表达水平显著下降(P<0.05)。Western blot法检测显示,I3C组与空白对照组比较,ERK总蛋白无明显变化(P>0.05),但p-ERK和Bcl-2蛋白表达显著减少(P<0.05),Bax蛋白表达和Caspase3活性显著增加(P<0.05)。【结论】吲哚-3-原醇能通过抑制ERK信号传导通路激活而抑制CNE-2细胞增值和促进CNE-2细胞凋亡。展开更多
文摘Beneficial effects of natural compounds are often attributed to modulation of NO production;however effects produced by plant extracts do not correlate with effects of purified components. The goal of our work was to study ability of flavonoids and indole-3-carbinol, as well as their combinations to modify NO production, iNOS gene and protein expression in rat tissues. Baicalein and luteolin decreased NO concentration in both intact and LPS-treated animals. Baicalein decreased iNOS gene expression. Luteolin decreased NO production in the liver and heart and number of iNOS-positive cells in the liver of LPS-treated animals. Combination of the two substances did not decrease the NO synthesis triggered by LPS, although it decreased iNOS gene expression. Quercetin decreased NO production in the heart, kidneys and blood of intact rats, but enhanced the LPS effect in testes, spleen and blood on NO production and iNOS protein expression in the liver. Indole-3-carbinol decreased NO concentration in the cerebellum, blood, lungs and skeletal muscles. The drug enhanced the LPS-triggered increase of NO production in all rat organs. It increased iNOS protein expression in intact liver;however it decreased the LPS-triggered outburst of the enzyme biosynthesis. Combination of indole3-carbinol with quercetin decreased NO production in LPS-treated animals however it slightly increased iNOS gene expression. Taken together our results suggest that modifications of NO level in tissues by a natural compound cannot be predicted from data about its effects on NOS expression or activity. Combination of substances can produce an effect differing from that of individual substances. This stresses importance of direct measurements of NO in the tissues.
基金supported by the National Natural Science Foundation of China(Grant Nos.:82073605 and 81803287)the Natural Science Foundation of Hebei Province(Grant Nos.:B2018201270 and H2021201002)+3 种基金the Talent Engineering Training Foundation of Hebei Province(Grant No.:A201802002)the Research Project of Hebei Provincial Administration of Traditional Chinese Medicine(Grant No.:2021174)Post-graduate's Innovation Fund Project of Hebei University(Grant No.:HBU2021ss006)the Outstanding Doctoral Cultivation Project of Hebei University(Grant No.:YB201703).
文摘Indole-3-carbinol(I3C),an important anticancer compound found in broccoli,has attracted considerable attention.The rapid extraction and accurate analysis of I3C in the pharmaceutical industry in broccoli is challenging as I3C is unstable at low pH and high temperature.In this study,a rapid,accurate,and lowcost ultrasound-assisted dispersive-filter extraction(UADFE)technique based on poly(deep eutectic solvent)-graphene oxide(PDES-GO)adsorbent was developed for the isolation and analysis of I3C in broccoli for the first time.PDES-GO with multiple adsorption interactions and a fast mass transfer rate was synthesized to accelerate adsorption and desorption.UADFE was developed by combining dispersive solid-phase extraction(DSPE)and filter solid-phase extraction(FSPE)to realize rapid extraction and separation.Based on the above two strategies,the proposed PDES-GO-UADFE method coupled with high-performance liquid chromatography(HPLC)allowed the rapid(15-16 min),accurate(84.3%-96.4%),and low-cost(adsorbent:3.00 mg)analysis of I3C in broccoli and was superior to solid-phase extraction,DSPE,and FSPE methods.The proposed method showed remarkable linearity(r=0.9998;range:0.0840-48.0 mg/g),low limit of quantification(0.0840 mg/g),and high precision(relative standard deviation<5.6%).Therefore,the PDES-GO-UADFE-HPLC method shows significant potential in the field of pharmaceutical analysis for the separation and analysis of anti-cancer compounds in complex plant samples.
文摘【目的】探讨吲哚-3-原醇(indol-3-carbinol,I3C)对人低分化鼻咽癌细胞株CNE-2的作用及其机制。【方法】将对数生长期的CNE-2细胞分成空白对照组和I3C组。空白对照组细胞常规培养,I3C组细胞培养体系中加入I3C(50μmol/L)。各组细胞培养48 h后,采用四甲基偶氮唑盐(MTT)法检测细胞增殖;Annexin V-碘化丙啶/异硫氰酸荧光素(PI/FITC)检测细胞凋亡,Real-time PCR检测细胞中胞外信号调节激酶(ERK)和Bax、Bcl-2基因表达;Western blot法检测细胞中ERK、p-ERK、Bax、Bcl-2蛋白表达,Caspase3活性试剂盒检测Caspase3活性。【结果】I3C组与空白对照组比较,CNE-2细胞增殖率显著下降(P<0.05),凋亡率显著升高(P<0.05),Real-time PCR检测显示I3C组与空白对照组比较,ERK m RNA表达无显著性差异,但Bax m RNA表达水平显著增高(P<0.001),Bcl-2 m RNA表达水平显著下降(P<0.05)。Western blot法检测显示,I3C组与空白对照组比较,ERK总蛋白无明显变化(P>0.05),但p-ERK和Bcl-2蛋白表达显著减少(P<0.05),Bax蛋白表达和Caspase3活性显著增加(P<0.05)。【结论】吲哚-3-原醇能通过抑制ERK信号传导通路激活而抑制CNE-2细胞增值和促进CNE-2细胞凋亡。
