目的:研究消癥散结方对高糖刺激下肾小球系膜细胞(MC)的保护作用以及对鞘氨醇激酶1/1磷酸鞘氨醇(Sphk1/S1P)信号通路的干预作用。方法:使用不同浓度消癥散结方含药血清和厄贝沙坦含药血清以及S1PR2抑制剂(JTE-013)与高糖共同培养MC,48h...目的:研究消癥散结方对高糖刺激下肾小球系膜细胞(MC)的保护作用以及对鞘氨醇激酶1/1磷酸鞘氨醇(Sphk1/S1P)信号通路的干预作用。方法:使用不同浓度消癥散结方含药血清和厄贝沙坦含药血清以及S1PR2抑制剂(JTE-013)与高糖共同培养MC,48h后流式细胞仪检测细胞周期,实时荧光定量PCR和Western Blot法检测FN、ColⅣ、TGF-β1、NF-κB(p65)、Sphk1和S1PR2的表达。结果:与高糖组比较,消癥散结方各剂量组细胞周期中G1期显著增加,S期显著减少(P<0.01);消癥散结方低剂量组细胞FN m RNA和NF-κB(p65)蛋白表达降低(P<0.05),ColⅣ蛋白、Sphk1 m RNA和蛋白表达显著降低(P<0.01);消癥散结方中剂量组细胞Sphk1m RNA表达显著降低(P<0.05),FN m RNA和Sphk1蛋白表达显著降低(P<0.01);消癥散结方高剂量组细胞,FN蛋白、NF-κB(p65)m RNA和蛋白表达显著降低(P<0.05),FN、ColⅣ、Sphk1与S1PR2的m RNA以及TGF-β1的m RNA和蛋白均显著降低(P<0.01)。结论:消癥散结方对高糖刺激下MC的保护作用可能与抑制Sphk1/S1P信号通路有关。展开更多
Osthole is observed to have the capacity to treat pulmonary arterial hypertension(PAH) in rats, but molecular mechanism is still unknown. The present study aims to discover therapeutic targets and explore therapeutic ...Osthole is observed to have the capacity to treat pulmonary arterial hypertension(PAH) in rats, but molecular mechanism is still unknown. The present study aims to discover therapeutic targets and explore therapeutic mechanism of osthole against PAH from metabolic perspective. A rat model with PAH was successfully established with MCT, following osthole administration, then untargeted metabolomics assay was performed using UPLC-Q-TOF-MS to identify differential metabolites and associated metabolic pathways, at last mechanism investigation was done by qRT-PCR, Western blot and ELISA. Differential metabolites characterized in rats with PAH were mostly assigned to sphingolipid metabolism, synthesis of unsaturated fatty acids, glycolysis, nucleotide metabolism, steroid hormone biosynthesis. Furthermore, osthole reversed high level of S1 P by modulating metabolic enzyme Sphk1 in rats with PAH. In addition, osthole inhibited the expression of Sphk1 by downregulating microRNA-21, phosphorylation of Akt, phosphorylation of mTOR in vivo and in vitro. These results demonstrated that metabolomics is a promising approach to discover potential drug target for PAH treatment. Importantly, our findings further elucidated therapeutic mechanism of osthole, a natural product, having a role of metabolic regulator to potentially treat PAH by targeting inhibition of Sphk1/S1 P via microRNA-21-PI3 K/Akt/mTOR signal pathway. Altogether, this discovery paves a critical foundation for enabling osthole to be a candidate compound to treat PAH.展开更多
文摘目的:研究消癥散结方对高糖刺激下肾小球系膜细胞(MC)的保护作用以及对鞘氨醇激酶1/1磷酸鞘氨醇(Sphk1/S1P)信号通路的干预作用。方法:使用不同浓度消癥散结方含药血清和厄贝沙坦含药血清以及S1PR2抑制剂(JTE-013)与高糖共同培养MC,48h后流式细胞仪检测细胞周期,实时荧光定量PCR和Western Blot法检测FN、ColⅣ、TGF-β1、NF-κB(p65)、Sphk1和S1PR2的表达。结果:与高糖组比较,消癥散结方各剂量组细胞周期中G1期显著增加,S期显著减少(P<0.01);消癥散结方低剂量组细胞FN m RNA和NF-κB(p65)蛋白表达降低(P<0.05),ColⅣ蛋白、Sphk1 m RNA和蛋白表达显著降低(P<0.01);消癥散结方中剂量组细胞Sphk1m RNA表达显著降低(P<0.05),FN m RNA和Sphk1蛋白表达显著降低(P<0.01);消癥散结方高剂量组细胞,FN蛋白、NF-κB(p65)m RNA和蛋白表达显著降低(P<0.05),FN、ColⅣ、Sphk1与S1PR2的m RNA以及TGF-β1的m RNA和蛋白均显著降低(P<0.01)。结论:消癥散结方对高糖刺激下MC的保护作用可能与抑制Sphk1/S1P信号通路有关。
基金the National Natural Science Foundation of China Grants (No. 81302764)the Science and Technology Grant for Excellent Talents of Harbin (No.2017RAXXJ060)。
文摘Osthole is observed to have the capacity to treat pulmonary arterial hypertension(PAH) in rats, but molecular mechanism is still unknown. The present study aims to discover therapeutic targets and explore therapeutic mechanism of osthole against PAH from metabolic perspective. A rat model with PAH was successfully established with MCT, following osthole administration, then untargeted metabolomics assay was performed using UPLC-Q-TOF-MS to identify differential metabolites and associated metabolic pathways, at last mechanism investigation was done by qRT-PCR, Western blot and ELISA. Differential metabolites characterized in rats with PAH were mostly assigned to sphingolipid metabolism, synthesis of unsaturated fatty acids, glycolysis, nucleotide metabolism, steroid hormone biosynthesis. Furthermore, osthole reversed high level of S1 P by modulating metabolic enzyme Sphk1 in rats with PAH. In addition, osthole inhibited the expression of Sphk1 by downregulating microRNA-21, phosphorylation of Akt, phosphorylation of mTOR in vivo and in vitro. These results demonstrated that metabolomics is a promising approach to discover potential drug target for PAH treatment. Importantly, our findings further elucidated therapeutic mechanism of osthole, a natural product, having a role of metabolic regulator to potentially treat PAH by targeting inhibition of Sphk1/S1 P via microRNA-21-PI3 K/Akt/mTOR signal pathway. Altogether, this discovery paves a critical foundation for enabling osthole to be a candidate compound to treat PAH.
