[Objectives]To optimize the determination method of oleandrin and adynerin in blood. [Methods]High performance liquid chromatography-mass spectrometry( HPLC-MS/MS) was applied to determine oleandrin and adynerin in bl...[Objectives]To optimize the determination method of oleandrin and adynerin in blood. [Methods]High performance liquid chromatography-mass spectrometry( HPLC-MS/MS) was applied to determine oleandrin and adynerin in blood. The blood sample was dispersed and fixed on a solid phase supported liquid-liquid extraction column and eluted with ethyl acetate. The resulting eluent was used for chromatographic separation with Kinetex C_(18) column as the separation column and gradient elution was performed using 10 mmol/L ammonium formate solution containing 0. 1%( volume fraction) formic acid and acetonitrile as the mobile phase. In the tandem mass spectrometry analysis,the detection was carried out using the electrospray positive ion source multiple reaction monitoring mode. [Results] The mass concentration of oleandrin and adynerin showed linear relationship in the range of 2-100 μg/L. The limit of detection( 3 S/N) of the method was 0. 5 μg/L.A blank sample was used as the substrate for the spike recovery test. The recovery rate was in the range of 90. 0%-98. 0%,and the relative standard deviation( RSD) of the measured values( n = 6) was in the range of 2. 1%-7. 3%. [Conclusions]The method established in this experiment has the benefits of simple pretreatment,good recovery,high sensitivity and strong specificity,and is expected to provide an ideal method for the determination of such drugs in blood.展开更多
Inflammation can initiate osteolysis,which is the breakdown of bone by fully developed osteoclasts.The com-pound Oleandrin is recognized for its effects against inflammation and tumors.Our objective was to examine the...Inflammation can initiate osteolysis,which is the breakdown of bone by fully developed osteoclasts.The com-pound Oleandrin is recognized for its effects against inflammation and tumors.Our objective was to examine the effects of Oleandrin on osteoclastogenesis and osteolysis,both in vitro and in vivo.In vitro,the impact of Oleandrin on osteoclastogenesis was assessed using CCK-8 assays,TRAP staining,and bone resorption assays.Ad-ditionally,a mouse model of osteolysis caused by LPS injection into the calvaria was used to conduct an in vivo investigation,examining bone histomorphology,histology,and immunohistochemistry.In vitro,concentrations of 5 nM and 10 nM of Oleandrin were found to be non-cytotoxic based on the results obtained.In vitro,Olean-drin hindered the osteoclastogenesis and bone resorption induced by RANKL.Oleandrin successfully inhibited the phosphorylation of NF-κB p65 and PI3K p85 in osteolytic tissue,thereby suppressing LPS-induced inflammatory osteolysis in mice calvaria during the in vivo study.Furthermore,the Oleandrin-treated group exhibited a note-worthy decrease in the expression level of NFATc1,which is a crucial controller of osteoclastogenesis.To sum up,our discoveries indicate that Oleandrin could hinder osteoclastogenesis and bone resorption,thereby having the ability to suppress inflammation-induced osteolysis.The underlying mechanism involves the NF-κB/PI3K pathway and inhibition of NFATc1 activation.Therefore,the findings suggest that Oleandrin holds potential as a therapeutic remedy for osteolytic ailments.展开更多
基金Supported by Project of National Natural Science Foundation(81273346)
文摘[Objectives]To optimize the determination method of oleandrin and adynerin in blood. [Methods]High performance liquid chromatography-mass spectrometry( HPLC-MS/MS) was applied to determine oleandrin and adynerin in blood. The blood sample was dispersed and fixed on a solid phase supported liquid-liquid extraction column and eluted with ethyl acetate. The resulting eluent was used for chromatographic separation with Kinetex C_(18) column as the separation column and gradient elution was performed using 10 mmol/L ammonium formate solution containing 0. 1%( volume fraction) formic acid and acetonitrile as the mobile phase. In the tandem mass spectrometry analysis,the detection was carried out using the electrospray positive ion source multiple reaction monitoring mode. [Results] The mass concentration of oleandrin and adynerin showed linear relationship in the range of 2-100 μg/L. The limit of detection( 3 S/N) of the method was 0. 5 μg/L.A blank sample was used as the substrate for the spike recovery test. The recovery rate was in the range of 90. 0%-98. 0%,and the relative standard deviation( RSD) of the measured values( n = 6) was in the range of 2. 1%-7. 3%. [Conclusions]The method established in this experiment has the benefits of simple pretreatment,good recovery,high sensitivity and strong specificity,and is expected to provide an ideal method for the determination of such drugs in blood.
基金National Natural Science Foundation of China(82272165)Shanghai Municipal Commission of Health and Family Planning’s Science and Research Fund(202040141)+2 种基金Shanghai Municipality Science and Technology Commission(20ZR1451800,22ZR1457200)Shanghai Municipal Health Commission(2022YQ006)Shanghai Tongren Hospital(TRKYRC-xx202203).
文摘Inflammation can initiate osteolysis,which is the breakdown of bone by fully developed osteoclasts.The com-pound Oleandrin is recognized for its effects against inflammation and tumors.Our objective was to examine the effects of Oleandrin on osteoclastogenesis and osteolysis,both in vitro and in vivo.In vitro,the impact of Oleandrin on osteoclastogenesis was assessed using CCK-8 assays,TRAP staining,and bone resorption assays.Ad-ditionally,a mouse model of osteolysis caused by LPS injection into the calvaria was used to conduct an in vivo investigation,examining bone histomorphology,histology,and immunohistochemistry.In vitro,concentrations of 5 nM and 10 nM of Oleandrin were found to be non-cytotoxic based on the results obtained.In vitro,Olean-drin hindered the osteoclastogenesis and bone resorption induced by RANKL.Oleandrin successfully inhibited the phosphorylation of NF-κB p65 and PI3K p85 in osteolytic tissue,thereby suppressing LPS-induced inflammatory osteolysis in mice calvaria during the in vivo study.Furthermore,the Oleandrin-treated group exhibited a note-worthy decrease in the expression level of NFATc1,which is a crucial controller of osteoclastogenesis.To sum up,our discoveries indicate that Oleandrin could hinder osteoclastogenesis and bone resorption,thereby having the ability to suppress inflammation-induced osteolysis.The underlying mechanism involves the NF-κB/PI3K pathway and inhibition of NFATc1 activation.Therefore,the findings suggest that Oleandrin holds potential as a therapeutic remedy for osteolytic ailments.
