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AIM:To investigate the possible mechanism of resveratrol(Res)on tumor necrosis factor-α (TNF-α)-induced monocyte chemoattractant protein-1(MCP-1)expression in primary rat pulmonary artery endothelial cells(RPAECs).METHODS: RPAECs were randomly divided into 4 groups: control group, solvent(1% DMSO) group,TNF-αgroup and Res group.Each group was divided into 1 h,4 h and 8 h subgroups(n=6 per time point).The TNF-α+C1142(a rodent chimeric mAb that neutralizes rat MCP-1)group was set up at the 8 h time point.At each time point,the protein and mRNA expression of MCP-1 was measured by Western blot and real-time PCR.RESULTS: Pre-treatment of the RPAECs with C1142 significantly down-regulated the expression of MCP-1(P<0.05).The protein and mRNA expression of MCP-1 was markedly increased in TNF-αgroup(P<0.05).Notably,incubation with Res down-re-gulated the protein and mRNA expression of MCP-1,which was significantly lower than that in TNF-αgroup(P<0.05). CONCLUSION:MCP-1 was involved in the process of TNF-α-induced injury of RPAECs.Res down-regulates the expres-sion of MCP-1 in RPAECs,thus attenuating cell injury.
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Inflammasome, which is a large multiprotein complex in the cytosol regulating IL-1β production, plays an important role in atherosclerosis ( AS ) . To date, NLRP3 ( nucleotide-binding domain and leucine-rich repeat protein 3) inflammasome is the most widely studied type of inflammasome. This article aims to review the effects of NLRP3 inflammasome on AS-related cells ( endothelial cells, macrophages and vascular smooth mus-cle cells) to further explore the role of NLRP3 inflammasome in the progression of AS.
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Aim To investigate the effect of ginkgolide B on TLR4 expression in glucose-treated endothelial cells.Methods Human umbilical vein endothelial cells (HUVECs)were stimulated by high concentra-tion of glucose.TLR4,inflammatory protein expression and Akt phosphorylation were analyzed by Western blot.Transcription factor NF-κB nuclear translocation was analyzed by immunofluorescence.Results The expression of TLR4 and PAF receptor was increased in high glucose-treated HUVECs. In contrast, both ginkgolide B and CV3988 dose-dependently decreased TLR4 and PAF receptor expression in high glucose-treated cells,respectively.Ginkgolide B decreased in-flammatory protein ICAM-1 ,VCAM-1 expression.Mo-reover,ginkgolide B potently abolished Akt phospho-rylation and NF-κB p65 nuclear translocation.Conclu-sion Ginkgolide B can reduce TLR4,PAF receptor, ICAM-1 and VCAM-1 expression in high dose of glu-cose-treated HUVECs,the mechanism might be linked to inhibition of Akt phosphorylation and NF-κB activa-tion.
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Aim To optimize extraction of polysaccha-rides from Dendrobium officinale ( DOP ) and investi-gate the effects of DOP on the expression of Bax and Bcl-2 in vascular endothelial cell of human umbilical vein ( HUVEC) by the induction of high sugar. Meth-ods Response surface methodology ( RSM ) was em-ployed to optimize the conditions for extraction of DOP on the basis of single factor experiment. The HUVEC cells were divided into the blank control group, the normal blood sugar group, the high sugar stimulation group and different drug concentration group plus the high sugar stimulation group. The growth condition of cells was examined by methyl thiazolyl tetrazolium ( MTT) , and RT-PCR was used to detect the expres-sion of Bcl-2 Assaciated X protein ( Bax ) and B-cell lymphoma-2 ( Bcl-2 ) in vascular endothelial cells after 24 and 48 hours. Results The optimal extraction time was 2 h and the optimal liquid-solid ratio was 15. 75∶1 ( mL/g ) , and the optimal extraction frequency was 2. 8. Under these optimized conditions, the extraction rate of polysaccharides was 25. 1%, while the experi-mental extraction rate was 24. 9%, which was highly fit with the regression model. Compared with normal sugar group, high sugar group reduced the growth of cells and enhanced the expression of Bax, and inhibi-ted the expression of Bcl-2 . DOP promoted the growth of cells in high sugar induction, inhibited the expres-sion of Bax, and enhanced the expression of Bcl-2. Conclusions The process optimized through RSM has the extraction effect, and has certain practical signifi-cance. Through the down-regulation of the expression of Bax in vascular endothelial cells and the up-regula-tion of the expression of Bcl-2 by the induction of high sugar, DOP can restrain HUVEC, which could prevent and cure diabetic angiopathy to some extent.
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Aim To investigate the effect of ginkgolide B on junctional proteins in ox-LDL-stimulated human umbilical vein endothelial cells ( HUVECs) . Methods After incubation with ginkgolide B ( 0 . 2 ,0 . 4 ,0 . 6 g · L-1 ) for 1 h, HUVECs were treated with ox-LDL (0. 1 g·L-1 ) for 4 h. The expressions of JAM-A and Cx43 were analyzed with Western blot and immunofluo-rescence. The effect of ginkgolide B on vascular per-meability was analyzed by Transwell experiments. Re-sults JAM-A and Cx43 expressions increased by 22%and 24% in ox-LDL-treated HUVECs, respectively. Whereas ginkgolide B significantly decreased JAM-A and Cx43 expressions. LY294002, a specific inhibitor of PI3K, suppressed JAM-A and Cx43 expressions in ox-LDL-stimulated cells. Ginkgolide B potently re-duced monocyte migration in ox-LDL-treated cells. Conclusion Ginkgolide B significantly suppresses JAM-A and Cx43 expressions, and reduces monocyte migration in ox-LDL-stimulated cells. This demon-strates that ginkgolide B can improve vascular permea-bility. The mechanism might be associated with the in-hibition of PI3K/Akt signaling pathway.