Investigate The Inhibition Effect Of Fetuin-A And MGP In CAP Induced MCP-1 And TGF-β Releasing And Expressing From Human Renal Tubulae Epithelial Cells Through The Regulation Of NF-KB Pathways

Objective:To investigate the inhibition effect of FetuinA and MGP in CaP induced MCP-1 and TGF-β releasing and expressing from human renal tubulae epithelial cells through the regulation of NF-KB pathways.Methods:1.The HK-2 was stimulated with 100ug/ml CaP,culture cells for 1、3、6、24、48、72h, The total RNA of the cultured cells was extracted, and then the NF-KB mRNA and MCP-1mRNA (1、3、 6h). TGF-βmRNA (24、48、72h) were detected by RT-PCR.2.The HK-2 was stimulated with 100ug/mlCaP+50mmol/L PDTC、 100ug/mlCaP+100mmol/L PDTC、100ug/mlCaP+200mmol/L PDTC respectively, culture cells for 6% 72h, The total RNA of the cultured cells was extracted, and then the MCP-1mRNA (6h)、TGF-βmRNA(72h) were detected by RT-PCR.3.The HK-2 was stimulated with 100ug/mlCaP+1000ng/ml FetuinA、100ug/mlCaP+2500ng/ml FetuinA、 100ug/mlCaP+5000ng/ml FetuinA.100ug/mlCaP+2000ng/ml MGP、 100ug/mlCaP+4000ng/ml MGP、100ug/mlCaP+6000ng/ml MGP respectively, culture cells for 3h, The total RNA of the cultured cells was extracted, and then the NF-KB (3h) was detected by RT-PCR.4. The HK-2 was stimulated with 100ug/ml CaP, culture cells for 1、3、6、24、48、 72h; the HK-2 was stimulated with 100ug/ml CaP+50mmol/L PDTC、 100ug/ml CaP+100mmol/L PDTC.100ug/ml CaP+200mmol/L PDTC respectively, culture cells for 6,72h, the cell culture supernatant was collected and MCP-1、TGF-β were determined by ELISA.Results:1.The NF-KBmRNA、MCP-1mRNA and TGF-βmRNA expression in 100ug/ml CaP were higher than in HK-2 alone. Peak expression of NF-KBmRNA occurred at 3h, Peak expression of MCP-1mRNA occurred at 6h, Peak expression of TGF-βmRNA occurred at72h (P<0.05) 2.The MCP-1mRNA and TGF-βmRNA expression in 100ug/mlCaP+50mmol/LPDTC、100ug/mlCaP+100mmol/L PDTC、 100ug/mlCaP+200mmol/L PDTC were lower than in 100ug/ml CaP alone. (P<0.05) 3.The NF-KBmRNA expression in 100ug/mlCaP+1000ng/mlFetuinA,100ug/mlCaP+2500ng/mlFetuinA、 100ug/mlCaP+5000ng/mlFetuinA.100ug/mlCaP+2000ng/mlMGP、 100ug/ml CaP+4000ng/mlMGP、100ug/mlCaP+6000ng/mlMGP were lower than in 100ug/ml CaP alone (P<0.05).4. The MCP-1 and TGF-βin the cell culture supernatant of 100ug/ml CaP was higher than in HK-2 alone (P<0.05). The MCP-1 and TGF-β in the cell culture supernatant of 100ug/ml CaP+50mmol/L PDTC.100ug/ml CaP+100mmol/L PDTC、 100ug/ml CaP+200mmol/L PDTC was lower than in 100ug/ml CaP (P< 0.05).Conclusions:1.stimulated the HK-2 by CaP was induced the release of inflammatory mediators, such as MCP-1、TGF-β, both may be play an important role in occurrence and development of Randall.2.The HK-2 was induced to the release of MCP-1 and TGF-βthroughthe activation of NF-KB possibly.3.FetuinA and MGP inhibited the expression of MCP-1 and TGF-β to decrease inflammation in the kidney damage, which may be the mechanism of kidney stone FetuinA and MGP take part in.