ABSTRACT
Aim To investigate the effect that catalpol intervenes macrophage polarization mediated by mouse mesangial cells(MMCs) stimulated by advanced glycation end products(AGEs).Methods RAW264.7 macrophages and MMCs were co-cultured in vitro and divided into model group(100 mg·L-1 AGEs), control group(100 mg·L-1 BSA), catalpol(0.1, 1.0, 10.0 μmol·L-1) group, and aminoguanidine(1.0 μmol·L-1) group which was set as positive control.After being incubated with catalpol for 1 h, MMCs were stimulated by AGEs for 23 h.The proliferation-inhibition rate of MMCs was measured by MTT assay.MCP-1 in supernatant liquid of MMCs was detected by ELISA method.The expression of iNOS, CD16/32, TNF-α, COX-2, CD206 and Arg-1 was detected by Western blot.Simultaneously, the percentage of iNOS and CD206 was also measured by flow cytometry.Results AGEs could increase the level of MCP-1 secreted by MMCs.The expression of iNOS, TNF-α, CD16/32 and COX-2 protein of macrophage was up-regulated after MMCs stimulated by AGEs, while the expression of CD206 and Arg-1 was down-regulated.After being intervened by catalpol, these effects could be reversed.All the changes were concentration-related.Conclusions Catalpol can inhibit macrophages M1-type polarization process and promote M2-type polarization, which may be mediated through MCP-1 secreted by MMCs after AGEs stimulation.Catalpol can ameliorate inflammation and relieve diabetic kidney injury.
ABSTRACT
Aim To observe the protective mechanism of loganinand morroniside ( active components in Cornus officinalis) on HUVEC injury induced by advanced glycation end products ( AGEs ) .Methods HUVECs were cultured in vitro and divided into control group , model group ( AGEs group ) , loganin group , morroni-side group and aminoguanidine group ( set as positive control).After being incubated with loganin and mor-roniside( final concentrations were 100,10,1 μmol?L-1 ) for 1 h, HUVECs were stimulated by AGEs of 200 mg? L-1 for 24 h.Then, the cell viability was measured by using MTT method .The supernatant was extracted and the levels of NO ,ET-1,MCP-1,VCAM-1 were measured by the corresponding kits .Receptors of advanced glycation end products ( RAGE ) and NF-κB in HUVEC were detected by Western blot .Results Loganin and morroniside could inhibit HUVEC injury induced by AGEs .In model group ,the contents of ET-1,MCP-1,VCAM-1 increased(P<0.01),the content of NO decreased ( P <0.01 ) and the expression of RAGE and NF-κB increased(P<0.01); however,lo-ganin and morronside could reduce the ET-1,MCP-1, VCAM-1contents,increase the NO content and down-regulate the expression of RAGE and NF-κB to differ-ent extents .Conclusion Loganin and morroniside could ameliorate HUVEC injury , and its mechanism may be related to inhibit inflammation , the improve-ment of endothelial cell function , and the decrease of the expression of RAGE .
ABSTRACT
Aim To explore the protective effect of lo-ganin ( an active component in Cornus officinalis ) on podocyte injury induced by advanced glycation end products ( AGEs) and its possible mechanism. Meth-ods Mouse podocytes were cultured in vitro and di-vided into Normal group, model group ( AGEs group) , loganin group and aminoguanidine group ( set as posi-tive control) . After being incubated with loganin( final concentrations are 0. 1, 1, 10 μmol · L-1 ) for 1 h, podocytes were stimulated by AGEs of 100 mg · L-1 for 24 h. Then, the cell viability was measured by u-sing MTT method. Podocytes apoptosis was evaluated by Hoechst33342/PI staining and flow cytometry. Re-ceptors of advanced glycation end products ( RAGE ) ,desmin and apoptosis-related protein like Bax, Bcl-2, cleaved caspase-3 in podocytes were detected by Western blot. Results Loganin ameliorated podocyte injury induced by AGEs, down-regulated the expression of desmin and RAGE. Loganin also reduced the apoptotic rate of podocytes and decreased the ratio of Bax/ Bcl-2 and the expression of pro-apoptotic protein cleaved caspase-3 in podocytes. Conclusion Loganin could ameliorate podocyte injury, and its mechanism may be related to the decrease of the expression of RAGE and inhibition of the apoptotic pathway.