Objective To study the therapeutic effects of Shenyuan Gan(参远苷,SYG)on the inflammat-ory response in BV2 microglial cells induced by lipopolysaccharide(LPS).Methods The cytotoxicity of SYG to BV2 microglial cells wa...Objective To study the therapeutic effects of Shenyuan Gan(参远苷,SYG)on the inflammat-ory response in BV2 microglial cells induced by lipopolysaccharide(LPS).Methods The cytotoxicity of SYG to BV2 microglial cells was evaluated using a Cell Counting Kit-8(CCK-8)assay,and the effect of SYG concentrations on LPS-induced BV2 microglial cells was studied.The morphological changes were observed using an optical microscope.The nitric oxide(NO)concentration in cell culture supernatant was determined using Griess re-agent.The expression of cytokines and inflammatory mediators were also measured by an en-zyme-linked immunosorbent assay(ELISA).Western blot analysis was used to determine the levels of inducible NO synthase(iNOS),nuclear factor-kappa B(NF-κB)p65,alpha inhibitor of NF-κB(IκB-α),phosphorylation-IκB-α(p-IκB-α),NOD-like receptor 3(NLRP3),and cas-pase-1 expression.Moreover,the expression of iNOS,NLRP3,and ionized calcium binding adapter molecule 1(Iba1)was also observed using immunofluorescent staining.Results SYG had a low cytotoxic effect on BV2 microglial cells and could significantly decr-ease LPS-induced morphological changes of BV2 microglial cells(P<0.05).ELISA results showed that SYG significantly inhibited the LPS-induced increase in interleukin(IL)-1βand IL-6 in BV2 microglia cells(P<0.05),and Western blot analysis showed that the phosphoryla-tion levels of iNOS,NF-κB p65,and IκB-αas well as NLRP3 and caspase-1 expression were also significantly decreased,and IκB-αexpression was increased after SYG treatment(P<0.05,compared with the LPS-treated group).The immunofluorescence results were consist-ent with the Western blot results,and Iba1 staining indicated that the cell morphology tended to be resting.These results indicate that SYG has a certain inhibitory effect on LPS-induced inflammation in BV2 microglial cells.Conclusion SYG can inhibit LPS-induced release of inflammatory factors in BV2 microglial cells by affecting the phosphorylation levels of NF-κB p65 and IκB-α.SYG is a valuable candid-ate for treating neuroinflammation-related diseases.展开更多
The objective of this study was to produce the porous col lagen-chitosan/Gl ycosanminglycans(GAG) for corneal cell-seed implant as a t hree-dimensional tissue engineering scaffold to improve the regeneration cornea s....The objective of this study was to produce the porous col lagen-chitosan/Gl ycosanminglycans(GAG) for corneal cell-seed implant as a t hree-dimensional tissue engineering scaffold to improve the regeneration cornea s.The effect of various content of glycerol as form porous agent to collagen-ch i tosan/GAG preserved a porous dimensional structure was investigated.The heat-dr ying was used to prepare porous collagen-chitosan /GAG scaffold.The pore morpho logy of collagen-chitosan/GAG was controlled by changing the concentration of g lycerol solution and drying methods.The porous structure morphology was observed by SEM.The diameter of the pores form 10 to 50 μm.The highly porous scaffold had interconnecting pores.The corneal cell morphology was observed under the li ght microscope.These results suggest that collagen-chitosan/GAG showed that cor neal cell have formed confluent layers and resemble the surface of normal cornea l cell surface.展开更多
基金The Space Medical Experiment Project of the China Manned Space Program(HYZHXM05003)National Natural Science Foundation of China(82171493)+2 种基金Natural Science Foundation of Hunan province(2021JJ30504)Scientific and Technological Innovation Project of the China Academy of Chinese Medical Sciences(CI2021A04905)Scientific Research Fund of Hunan Provincial Education of the Hunan University of Traditional Chinese Medicine First-class Discipline Project of Chinese Medicine(19B422)。
文摘Objective To study the therapeutic effects of Shenyuan Gan(参远苷,SYG)on the inflammat-ory response in BV2 microglial cells induced by lipopolysaccharide(LPS).Methods The cytotoxicity of SYG to BV2 microglial cells was evaluated using a Cell Counting Kit-8(CCK-8)assay,and the effect of SYG concentrations on LPS-induced BV2 microglial cells was studied.The morphological changes were observed using an optical microscope.The nitric oxide(NO)concentration in cell culture supernatant was determined using Griess re-agent.The expression of cytokines and inflammatory mediators were also measured by an en-zyme-linked immunosorbent assay(ELISA).Western blot analysis was used to determine the levels of inducible NO synthase(iNOS),nuclear factor-kappa B(NF-κB)p65,alpha inhibitor of NF-κB(IκB-α),phosphorylation-IκB-α(p-IκB-α),NOD-like receptor 3(NLRP3),and cas-pase-1 expression.Moreover,the expression of iNOS,NLRP3,and ionized calcium binding adapter molecule 1(Iba1)was also observed using immunofluorescent staining.Results SYG had a low cytotoxic effect on BV2 microglial cells and could significantly decr-ease LPS-induced morphological changes of BV2 microglial cells(P<0.05).ELISA results showed that SYG significantly inhibited the LPS-induced increase in interleukin(IL)-1βand IL-6 in BV2 microglia cells(P<0.05),and Western blot analysis showed that the phosphoryla-tion levels of iNOS,NF-κB p65,and IκB-αas well as NLRP3 and caspase-1 expression were also significantly decreased,and IκB-αexpression was increased after SYG treatment(P<0.05,compared with the LPS-treated group).The immunofluorescence results were consist-ent with the Western blot results,and Iba1 staining indicated that the cell morphology tended to be resting.These results indicate that SYG has a certain inhibitory effect on LPS-induced inflammation in BV2 microglial cells.Conclusion SYG can inhibit LPS-induced release of inflammatory factors in BV2 microglial cells by affecting the phosphorylation levels of NF-κB p65 and IκB-α.SYG is a valuable candid-ate for treating neuroinflammation-related diseases.
文摘The objective of this study was to produce the porous col lagen-chitosan/Gl ycosanminglycans(GAG) for corneal cell-seed implant as a t hree-dimensional tissue engineering scaffold to improve the regeneration cornea s.The effect of various content of glycerol as form porous agent to collagen-ch i tosan/GAG preserved a porous dimensional structure was investigated.The heat-dr ying was used to prepare porous collagen-chitosan /GAG scaffold.The pore morpho logy of collagen-chitosan/GAG was controlled by changing the concentration of g lycerol solution and drying methods.The porous structure morphology was observed by SEM.The diameter of the pores form 10 to 50 μm.The highly porous scaffold had interconnecting pores.The corneal cell morphology was observed under the li ght microscope.These results suggest that collagen-chitosan/GAG showed that cor neal cell have formed confluent layers and resemble the surface of normal cornea l cell surface.
