Investigate The Inhibition Effect Of Fetuin-A In Cap Induced HMGB1Releasing And Expression From Human Macrophage

Objective: To investigate the inhibition effect of CaP induced HMGB1releasing and expressing from human macrophage.Methods：The induced U937was stimulated with100ug/ml CaP,100ug/mlCaP+1000ng/ml fetuin-A,100ug/ml CaP+2500ng/ml fetuin-A,100ug/mlCaP+5000ng/ml fetuin-A respectively, culture cells for24h. The total RNA ofthe cultured cells was extracted，and then the HMGB1mRNA was detected byRT-PCR. After the total protein，the cytoplasm protein and nucleus protein beingextracted，the contents of HMGB1protein was detected by western blot.Thecell culture supernatant was collected,in which HMGB1protein was detectedby western blot；When each group cells were cultured for4h，The cell culturesupernatant was collected and IL-1β,IL-6,TNF-α,MCP-1was determined byELISA.Results：The HMGB1mRNA expression in100ug/ml CaP+1000ng/ml fetuin-A,100ug/ml CaP+2500ng/ml fetuin-A,100ug/ml CaP+5000ng/ml fetuin-A waslower than in CaP alone (P <0.05); the contents of HMGB1protein in the total protein and the cytoplasm of100ug/ml CaP+1000ng/ml fetuin-A,100ug/mlCaP+2500ng/ml fetuin-A,100ug/ml CaP+5000ng/ml fetuin-A was lower thanin CaP alone(P <0.05),while the nucleus protein of100ug/ml CaP+1000ng/mlfetuin-A,100ug/ml CaP+2500ng/ml fetuin-A,100ug/ml CaP+5000ng/mlfetuin-A was higher than in CaP alone (P <0.05); The HMGB1protein and IL-1β,IL-6,TNF-α and MCP-1in the cell culture supernatant of100ug/mlCaP+1000ng/mlfetuin-A,100ug/mlCaP+2500ng/ml fetuin-A,100ug/mlCaP+5000ng/ml fetuin-A was lower than in CaP alone (P <0.05);Conclusions：1. Fetuin-A suppressed the expression of HMGB1mRNA in macrophagesinduced by CaP; Fetuin-A inhibited the synthesis of HMGB1protein inmacrophage induced by CaP;2. Fetuin-A inhibited the transposition and release of HMGB1protein inmacrophage induced by CaP; Fetuin-A decreased the level of HMGB1proteinin macrophages cell culture supernatant;3. Fetuin-A inhibited the release of IL-1β、IL-6、 TNF-α、MCP-1inmacrophages induced by CaP; Fetuin-A decreased the level of IL-1β、IL-6、TNF-α、MCP-1in the macrophages cell culture supernatant;...