Nicotinicα4β2 receptor antagonists have drawn increasing attention in the development of new antidepressants.In this study,we aimed to investigate the protective effect of VMY-2-95,the new selective antagonist ofα4...Nicotinicα4β2 receptor antagonists have drawn increasing attention in the development of new antidepressants.In this study,we aimed to investigate the protective effect of VMY-2-95,the new selective antagonist ofα4β2 nicotinic acetylcholine receptor(nAChR)on corticosterone(CORT)injured mice and cellular models.Fluoxetine was applied as a positive control,and the effects of VMY-2-95 were investigated with three different doses or concentrations(1,3,10 mg/kg in mice,and 0.003,0.03,0.1μmol/L in cells).As a result,VMY-2-95 showed significant antidepressant-like effects in the CORT injured mice by improving neuromorphic function,promoting hippocampal nerve proliferation,and regulating the contents of monoamine transmitters.Meanwhile,VMY-2-95 exhibited protective effects on cell viability,cell oxidant,cell apoptosis,and mitochondrial energy metabolism on corticosterone-impaired SH-SY5 Y cells.Also,the PKA-CREB-BDNF signaling pathway was up-regulated by VMY-2-95 both in vitro and in vivo,and pathway blockers were also combined with VMY-2-95 to verify the effects furtherly.Therefore,we preliminarily proved that VMY-2-95 had protective effects in depressed mice and SH-SY5 Y cells against injuries induced by corticosterone.This work indicated that the application of VMY-2-95 is a potential pharmacological solution for depression.This study also supported the development ofα4β2 nAChR antagonists towards neuropsychiatric dysfunctions.展开更多
[目的]探讨miR-552-3p与PTEN/AKT信号通路对子宫内膜癌细胞生物活性的作用机制。[方法]将RL95-2细胞分为两组:inhibitor NC组(NC组)和miR-552-3p inhibitor组(inhibitor组);或Si NC组和Si PTEN组。通过MTT检测细胞活性,流式细胞术检测...[目的]探讨miR-552-3p与PTEN/AKT信号通路对子宫内膜癌细胞生物活性的作用机制。[方法]将RL95-2细胞分为两组:inhibitor NC组(NC组)和miR-552-3p inhibitor组(inhibitor组);或Si NC组和Si PTEN组。通过MTT检测细胞活性,流式细胞术检测细胞凋亡率,Transwell小室检测细胞侵袭数目,划痕实验检测细胞迁移距离,免疫印迹检测细胞相关蛋白水平。[结果]与hEEC细胞比较,miR-552-3p在肿瘤细胞表达异常升高(0.62±0.03 vs 1.56±0.07);和转染inhibitor NC相比,转染miR-552-3p inhibitor后,RL95-2细胞的活性降低,RL95-2细胞的迁移和侵袭能力显著降低(25.29%±2.89%vs 11.27%±1.23%;65.33±3.12 vs 40.27±8.03),PTEN蛋白表达降低(0.93±0.12 vs 0.67±0.03),CCS-3和Bax的表达增加(0.97±0.11 vs 1.72±0.09,1.01±0.10 vs3.03±0.09)。和转染Si NC相比,转染Si PTEN后,RL95-2细胞的活性被明显抑制,细胞凋亡明显增加(11.02%±0.35%vs 30.89%±1.23%),CCS-3和Bax的表达增加(0.23±0.21 vs 1.09±0.08,0.21±0.31 vs 0.82±0.05)。双荧光素酶报告基因实验表明,相较于inhibitor NC组细胞,miR-552-3p innhibitor组细胞PTEN-WT报告基因的荧光素酶活性显著降低(0.93±0.09 vs 0.59±0.05)。[结论]抑制miR-552-3p能够抑制子宫内膜癌细胞的生物活性,这一过程与miR-552-3p调控PTEN/AKT通路有关。展开更多
Galectin-3(gal-3)and its ligands have been implicated in cell transformation and cancer metastasis.Gal-3 protein has been found in uterine epithelial cells adjacent to implanting blastocysts in different cell types.In...Galectin-3(gal-3)and its ligands have been implicated in cell transformation and cancer metastasis.Gal-3 protein has been found in uterine epithelial cells adjacent to implanting blastocysts in different cell types.In order to investigate the role of gal-3 in the establishment of human endometrial receptivity,the expression of gal-3 in human endometrial cell line RL95-2 was silenced by RNA inter-ference technology using gal-3 specific small RNA.The expression of gal-3 was detected by the reverse transcriptase-polymerase chain reaction and Western blot analysis.After the suppression of gal-3,cell cycle changes and the expres-sion of integrin b1 were detected by flow cytometry.The adhesive ability of RL95-2 cells was analyzed by the adhesion test.Gal-3 siRNA transfection efficiency reached 70%–90%.The expression of gal-3 mRNA and protein in RL95-2 cells was strongly inhibited by 70%–90%after RNA interference.Inhibition of gal-3 expression decreased S-phase but increased G1 phase cells.Integrin b1 expression was down-regulated,and the adhesive ability of RL95-2 cells to fibronectin(FN)was significantly reduced.Gal-3 may be involved in the establishment of endometrial receptivity by regulating the proliferation and adhesion of endometrial cells.The influence on adhesion may be related with the integrin modulation.展开更多
基金supported by the National Natural Science Foundation of China(No.81603100)the Drug Innovation Major Project(2018ZX09711001-003-005,2018ZX09711001-012,China)+1 种基金CAMS Innovation Fund for Medical Sciences(2017-I2M1-010,China)Peking Union Medical College Graduate Innovation Fund Project(2017-1007-11,China)
文摘Nicotinicα4β2 receptor antagonists have drawn increasing attention in the development of new antidepressants.In this study,we aimed to investigate the protective effect of VMY-2-95,the new selective antagonist ofα4β2 nicotinic acetylcholine receptor(nAChR)on corticosterone(CORT)injured mice and cellular models.Fluoxetine was applied as a positive control,and the effects of VMY-2-95 were investigated with three different doses or concentrations(1,3,10 mg/kg in mice,and 0.003,0.03,0.1μmol/L in cells).As a result,VMY-2-95 showed significant antidepressant-like effects in the CORT injured mice by improving neuromorphic function,promoting hippocampal nerve proliferation,and regulating the contents of monoamine transmitters.Meanwhile,VMY-2-95 exhibited protective effects on cell viability,cell oxidant,cell apoptosis,and mitochondrial energy metabolism on corticosterone-impaired SH-SY5 Y cells.Also,the PKA-CREB-BDNF signaling pathway was up-regulated by VMY-2-95 both in vitro and in vivo,and pathway blockers were also combined with VMY-2-95 to verify the effects furtherly.Therefore,we preliminarily proved that VMY-2-95 had protective effects in depressed mice and SH-SY5 Y cells against injuries induced by corticosterone.This work indicated that the application of VMY-2-95 is a potential pharmacological solution for depression.This study also supported the development ofα4β2 nAChR antagonists towards neuropsychiatric dysfunctions.
文摘[目的]探讨miR-552-3p与PTEN/AKT信号通路对子宫内膜癌细胞生物活性的作用机制。[方法]将RL95-2细胞分为两组:inhibitor NC组(NC组)和miR-552-3p inhibitor组(inhibitor组);或Si NC组和Si PTEN组。通过MTT检测细胞活性,流式细胞术检测细胞凋亡率,Transwell小室检测细胞侵袭数目,划痕实验检测细胞迁移距离,免疫印迹检测细胞相关蛋白水平。[结果]与hEEC细胞比较,miR-552-3p在肿瘤细胞表达异常升高(0.62±0.03 vs 1.56±0.07);和转染inhibitor NC相比,转染miR-552-3p inhibitor后,RL95-2细胞的活性降低,RL95-2细胞的迁移和侵袭能力显著降低(25.29%±2.89%vs 11.27%±1.23%;65.33±3.12 vs 40.27±8.03),PTEN蛋白表达降低(0.93±0.12 vs 0.67±0.03),CCS-3和Bax的表达增加(0.97±0.11 vs 1.72±0.09,1.01±0.10 vs3.03±0.09)。和转染Si NC相比,转染Si PTEN后,RL95-2细胞的活性被明显抑制,细胞凋亡明显增加(11.02%±0.35%vs 30.89%±1.23%),CCS-3和Bax的表达增加(0.23±0.21 vs 1.09±0.08,0.21±0.31 vs 0.82±0.05)。双荧光素酶报告基因实验表明,相较于inhibitor NC组细胞,miR-552-3p innhibitor组细胞PTEN-WT报告基因的荧光素酶活性显著降低(0.93±0.09 vs 0.59±0.05)。[结论]抑制miR-552-3p能够抑制子宫内膜癌细胞的生物活性,这一过程与miR-552-3p调控PTEN/AKT通路有关。
基金supported by the National Natural Science Foundation of China(Grant No.30271354).
文摘Galectin-3(gal-3)and its ligands have been implicated in cell transformation and cancer metastasis.Gal-3 protein has been found in uterine epithelial cells adjacent to implanting blastocysts in different cell types.In order to investigate the role of gal-3 in the establishment of human endometrial receptivity,the expression of gal-3 in human endometrial cell line RL95-2 was silenced by RNA inter-ference technology using gal-3 specific small RNA.The expression of gal-3 was detected by the reverse transcriptase-polymerase chain reaction and Western blot analysis.After the suppression of gal-3,cell cycle changes and the expres-sion of integrin b1 were detected by flow cytometry.The adhesive ability of RL95-2 cells was analyzed by the adhesion test.Gal-3 siRNA transfection efficiency reached 70%–90%.The expression of gal-3 mRNA and protein in RL95-2 cells was strongly inhibited by 70%–90%after RNA interference.Inhibition of gal-3 expression decreased S-phase but increased G1 phase cells.Integrin b1 expression was down-regulated,and the adhesive ability of RL95-2 cells to fibronectin(FN)was significantly reduced.Gal-3 may be involved in the establishment of endometrial receptivity by regulating the proliferation and adhesion of endometrial cells.The influence on adhesion may be related with the integrin modulation.
