Objective:To investigate the effect of astragalus polysaccharide(APS)on macrophage phagocytosis of P.aerugino-sa and its related mechanism.Methods:The effect of APS on the activity of macrophages was detected by CCK-8 assay.The effect of APS on macrophage phagocytosis related signal pathways were detected by Western blot.Gentamicin protection test and flow cytometry were employed to evaluate the effect of APS on macrophage phagocytosis of P.aeruginosa.Then the macrophages were pretreated with the inhibitors of TLR4,NF-κB,STAT3,MAPK and PI3K to assess the mechanism mediating APS on macrophage phagocytosis.Results:APS promotes the activity of macrophages,and significantly promoted the phagocytosis of P.aeruginosa by macrophages.Fur-thermore,macrophage phagocytosis of P.aeruginosa was significantly inhibited by TLR4 inhibitors(TAK242)and PI3K inhibitors(LY294002),but not affected by NF-κB,STAT3 and MAPK inhibitors.Conclusion:APS has no toxicity on macrophages,and can promote the phagocytosis of macrophages to P.aeruginosa,the underlying mechanism may be related to TLR4/PI3K pathway.

Objective:To investigate the effect of astragalus polysaccharide(APS)on macrophage phagocytosis of P.aerugino-sa and its related mechanism.Methods:The effect of APS on the activity of macrophages was detected by CCK-8 assay.The effect of APS on macrophage phagocytosis related signal pathways were detected by Western blot.Gentamicin protection test and flow cytometry were employed to evaluate the effect of APS on macrophage phagocytosis of P.aeruginosa.Then the macrophages were pretreated with the inhibitors of TLR4,NF-κB,STAT3,MAPK and PI3K to assess the mechanism mediating APS on macrophage phagocytosis.Results:APS promotes the activity of macrophages,and significantly promoted the phagocytosis of P.aeruginosa by macrophages.Fur-thermore,macrophage phagocytosis of P.aeruginosa was significantly inhibited by TLR4 inhibitors(TAK242)and PI3K inhibitors(LY294002),but not affected by NF-κB,STAT3 and MAPK inhibitors.Conclusion:APS has no toxicity on macrophages,and can promote the phagocytosis of macrophages to P.aeruginosa,the underlying mechanism may be related to TLR4/PI3K pathway.

Objective To investigate the impact of long non-coding RNA(LncRNA)taurine up-regulated gene 1(TUG1)on high glucose-induced cardiomyocyte apoptosis by regulating miR-181b-5p/programmed cell death protein 4(PDCD4)axis.Methods Diabetic cardiomyopathy(DCM)cell model was established in vitro with high glucose(HG,25 mmol/L glucose).AC16 cells were divided into the NG(5.5 mmol/L glucose)group,the HG group,the HG+sh-NC group,the HG+sh-TUG1 group,the HG+miR-NC group,the HG+miR-181b-5p group,the HG+sh-TUG1+anti-miR-NC group,the HG+sh-TUG1+anti-miR-181b-5p group,the HG+miR-181b-5p+pcDNA group and HG+miR-181b-5p+pc-PDCD4 group.The Cell Counting Kit-8(CCK-8)method was applied to detect cell viability.Lactate dehydrogenase(LDH)assay was applied to detect LDH release.Quantitative real-time polymerase chain reaction(qRT-PCR)was applied to detect expression levels of TUG1,miR-181b-5p and PDCD4 mRNA.Flow cytometry was applied to detect apoptosis.Western blot assay was applied to detect levels of B-cell lymphoma 2-associated X(Bax),activated caspase 3(cleaved caspase 3)and PDCD4 proteins.Caspase-Glo3 assay was applied to assess caspase 3 activity.Dual-luciferase reporter assay was applied to verify the targeting relationship between TUG1 or PDCD4 and miR-181b-5p.Results Compared with the NG group,the cell activity decreased in the HG group,and LDH release,apoptosis rate,Bax,cleaved caspase 3 expression and caspase 3 activity increased(P＜0.05),which could be antagonized by TUG1 knockdown or miR-181b-5p overexpression(P＜0.05).Inhibition of miR-181b-5p was able to alleviate the impact of TUG1 silencing on cardiomyocyte viability and apoptosis under high glucose treatment(P＜0.05).The overexpression of PDCD4 attenuated the promotion effect of miR-181b-5p up-regulation on the viability of cardiomyocytes treated with high glucose and the inhibitory effect on apoptosis.TUG1 was able to increase the expression of PDCD4 through adsorption of miR-181b-5p(P＜0.05).Conclusion TUG1 promotes high glucose-induced cardiomyocyte apoptosis by down-regulating miR-181b-5p and up-regulating PDCD4.

Objective:Mesenchymal stem cells( MSCs) have self-renewal capacity and potential to differentiate into the cells.It was reported that the expression of miR-30a changed in some immune diseases.But it remains unclear the effect of miR-30a on the im-munoregulatory functions of MSCs.Here we studied the impact of miR-30a on the phenotype,cell viability,apoptosis,cell cycle and im-munoregulatory functions of MSCs.Methods: The mixed enzyme methods were used for the isolation of human umbilical cord MSCs.Flow cytometry(FCM)was used to investigate the effect of overexpressed miR-30a on the phenotype of MSCs.CCK-8 was used to examine the cell viability of miR-30a-overexpressed MSCs.Annexin V/PI was used for the detection of apoptosis of MSCs.Q-PCR and Western blot were used to investigate the effect of miR-30a on the expression of Cyclin E2( CCNE2).CCNE2 was one putative target of miR-30a predicted by Targetscan database.The effects of miR-30a-overexpressed MSCs on the maturation of dendritic cells(DCs)were determined.Results:Overexpression of miR-30a blocked the cell cycle of MSCs in the G0/G1 phase by inhibiting the expression of CCNE2,but did not affect the phenotype, cell viability and apoptosis of MSCs.When co-cultured with DCs, although MSCs down-regulated the expression of CD40 and CD86 on DCs,overexpression of miR-30a more significantly enhanced the suppressive impact of MSCs on the maturation of DCs.Conclusion: miR-30a affects the cell cycle of MSCs and enhances its immunosuppressive effect on DCs.