Objective:To investigate the effect and mechanism of miRNA-24 on the proliferation,migration and tube formation of vascular endothelial cells.Methods:Human umbilical vein endothelial cells(HUVECs)were assigned into co...Objective:To investigate the effect and mechanism of miRNA-24 on the proliferation,migration and tube formation of vascular endothelial cells.Methods:Human umbilical vein endothelial cells(HUVECs)were assigned into control,microRNA(miR)-24 overexpression and anti-miR-24 groups.The proliferation and migration abilities of HUVECs were detected by methyl thiazolyl tetrazolium(MTT)assay and scratch wound healing assay,respectively.The ability of HUVECs to form tubular structures was evaluated by a tube formation assay.The mRNA and protein expressions of vascular endothelial growth factor(VEGF)and transcription factor Sp1 were determined by RT-PCR,immunocytochemistry and western blotting,respectively.Results:The miR-24 overexpression group exhibited decreased cell proliferation and migration,and expressions of VEGF and Sp1 compared with the control group(P <0.01).No tube-like network structure was formed in the miR-24 overexpression group.However,inhibition of miR-24 in HUVECs markedly increased cell proliferation and migration,enhanced tube formation and expressions of VEGF and Sp1(P<0.05 or P<0.01).Conclusion:MiR-24 suppressed the proliferation,migration and tube formation of HUVECs,and the mechanism might be related to the down-regulation of VEGF expression.Sp1 might participate in this regulation process.展开更多
Objective:To investigate the effects of miR-24 on the expression of vascular endothelial nitric oxide synthase(eNOS)and plasma nitric oxide(NO)level in rats.Methods:The miR-24 over-expression plasmid was constructed.T...Objective:To investigate the effects of miR-24 on the expression of vascular endothelial nitric oxide synthase(eNOS)and plasma nitric oxide(NO)level in rats.Methods:The miR-24 over-expression plasmid was constructed.Twenty healthy male Sprague-Dawley rats were randomly divided into two groups:miR-24 group and blank plasmid group.Rats were injected with plasmid DNA in saline via the tail vein.After eight weeks,the rat artery was removed and HE staining was used to observe the morphological changes.The protein expression and activity of eNOS were detected by immunohistochemistry and enzyme-linked immunosorbent assay(ELISA),respectively,and the NO level in plasma was determined by nitrate reduction assay.Results:Compared with the blank plasmid group,the expression and activity of eNOS as well as the NO production in the miR-24 group was significantly decreased(P <0.05 or P <0.01).No significant difference in the morphology of vascular tissue was noted between the two groups.Conclusion:Up-regulation of miR-24 inhibits the expression of eNOS and NO production in rat vascular tissue.It may provide a bio-target for the development of drugs to treat cardiovascular diseases such as hypertension.展开更多
[Objectives]This paper aims to establish a method to simultaneously determine the content ofβ-asarone in Rhizoma Acori Tatarinowii dried by three different methods.[Methods]Reversed-phase high-performance liquid chro...[Objectives]This paper aims to establish a method to simultaneously determine the content ofβ-asarone in Rhizoma Acori Tatarinowii dried by three different methods.[Methods]Reversed-phase high-performance liquid chromatography was used,and the chromatographic conditions were as follows:column,Thermo SCIENTIFIC Hypersil GOLD Dim.(mm);mobile phase,methanol-0.1%phosphoric acid(63∶37);column temperature,30℃;flow rate,1mL/min;detection wavelength,257 nm;sample size,10μL.[Results]The linear range of the injection volume ofβ-asarone was 49.28-246.40μg/mL(R=0.9993);the limit of quantification was 0.85 ng and the detection limit was 0.34 ng;the RSD values of precision,stability and reproducibility tests were all less than 3%;and the sample recovery rate was 98.53%-98.97%(RSD<3.00).The results show that the content ofβ-asarone was highest in shade-dried Rhizoma Acori Tatarinowii.The order ofβ-asarone content was as follows:Rhizoma Acori Tatarinowii dried in shade>Rhizoma Acori Tatarinowii dried at 55℃>Rhizoma Acori Tatarinowii dried at 60℃.[Conclusions]This method is sensitive,reliable,and reproducible.It can be used to simultaneously determine the content ofβ-asarone in Rhizoma Acori Tatarinowii dried by three different methods.展开更多
基金supported by the National Natural Science Foundation of China(No.81373403)
文摘Objective:To investigate the effect and mechanism of miRNA-24 on the proliferation,migration and tube formation of vascular endothelial cells.Methods:Human umbilical vein endothelial cells(HUVECs)were assigned into control,microRNA(miR)-24 overexpression and anti-miR-24 groups.The proliferation and migration abilities of HUVECs were detected by methyl thiazolyl tetrazolium(MTT)assay and scratch wound healing assay,respectively.The ability of HUVECs to form tubular structures was evaluated by a tube formation assay.The mRNA and protein expressions of vascular endothelial growth factor(VEGF)and transcription factor Sp1 were determined by RT-PCR,immunocytochemistry and western blotting,respectively.Results:The miR-24 overexpression group exhibited decreased cell proliferation and migration,and expressions of VEGF and Sp1 compared with the control group(P <0.01).No tube-like network structure was formed in the miR-24 overexpression group.However,inhibition of miR-24 in HUVECs markedly increased cell proliferation and migration,enhanced tube formation and expressions of VEGF and Sp1(P<0.05 or P<0.01).Conclusion:MiR-24 suppressed the proliferation,migration and tube formation of HUVECs,and the mechanism might be related to the down-regulation of VEGF expression.Sp1 might participate in this regulation process.
基金supported by the National Natural Science Foundation of China(No. 03101213068D)Guangxi Master Postgraduate Research Innovation Project(No.YCSZ2015119)
文摘Objective:To investigate the effects of miR-24 on the expression of vascular endothelial nitric oxide synthase(eNOS)and plasma nitric oxide(NO)level in rats.Methods:The miR-24 over-expression plasmid was constructed.Twenty healthy male Sprague-Dawley rats were randomly divided into two groups:miR-24 group and blank plasmid group.Rats were injected with plasmid DNA in saline via the tail vein.After eight weeks,the rat artery was removed and HE staining was used to observe the morphological changes.The protein expression and activity of eNOS were detected by immunohistochemistry and enzyme-linked immunosorbent assay(ELISA),respectively,and the NO level in plasma was determined by nitrate reduction assay.Results:Compared with the blank plasmid group,the expression and activity of eNOS as well as the NO production in the miR-24 group was significantly decreased(P <0.05 or P <0.01).No significant difference in the morphology of vascular tissue was noted between the two groups.Conclusion:Up-regulation of miR-24 inhibits the expression of eNOS and NO production in rat vascular tissue.It may provide a bio-target for the development of drugs to treat cardiovascular diseases such as hypertension.
基金Key Research and Development Project of Department of Science and Technology of Guangxi Zhuang Autonomous Region(AB19110003)Project for Improving Basic Scientific Research Ability of Young and Middle-aged Teachers in Colleges and Universities of Guangxi(2019KY0341,2019ky0344)+2 种基金Open Project of Guangxi Zhuang Yao Medicine Center of Engineering and Technology(KJT1900105)Youth Foundation of Guangxi University of Chinese Medicine(2019QN036,2019QN030)Traditional Chinese Medicine Scientific Research Laboratory(Grade III)of National Administration of Traditional Chinese Medicine:Laboratory of Chinese(Zhuang)Medicine Chemical and Quality Analysis(Guo Zhong Yi Yao Fa 2009[21]).
文摘[Objectives]This paper aims to establish a method to simultaneously determine the content ofβ-asarone in Rhizoma Acori Tatarinowii dried by three different methods.[Methods]Reversed-phase high-performance liquid chromatography was used,and the chromatographic conditions were as follows:column,Thermo SCIENTIFIC Hypersil GOLD Dim.(mm);mobile phase,methanol-0.1%phosphoric acid(63∶37);column temperature,30℃;flow rate,1mL/min;detection wavelength,257 nm;sample size,10μL.[Results]The linear range of the injection volume ofβ-asarone was 49.28-246.40μg/mL(R=0.9993);the limit of quantification was 0.85 ng and the detection limit was 0.34 ng;the RSD values of precision,stability and reproducibility tests were all less than 3%;and the sample recovery rate was 98.53%-98.97%(RSD<3.00).The results show that the content ofβ-asarone was highest in shade-dried Rhizoma Acori Tatarinowii.The order ofβ-asarone content was as follows:Rhizoma Acori Tatarinowii dried in shade>Rhizoma Acori Tatarinowii dried at 55℃>Rhizoma Acori Tatarinowii dried at 60℃.[Conclusions]This method is sensitive,reliable,and reproducible.It can be used to simultaneously determine the content ofβ-asarone in Rhizoma Acori Tatarinowii dried by three different methods.
