| Objective1.To investigate the regulatory effect of Clq on the expression LAIR-1 in THP-1 cells infected with H.pylori.2.To study the effect of LAIR-1 ligand C1q on the secretion of IL-4 IL-8 and IL-10 in THP-1 cells infected with H.pylori.3.To study the effect of Clq on the key molecules of PI3K/Akt,p38 MAPK,JNK and NF-?B signaling pathways in H.pylori infection,and to explore the relationship between these signaling pathways and the secretion of Clq regulated cytokines.Methods50 ?g/ml Clq and 1×105 monocyte THP-1 are co-culturing 24 h.And then co-cultured with H.pylori(MOI for 100:1)for 24 h.Cell supernatant is collected and IL-4,IL-8 or IL-10 are detected by ELISA,50 ?g/ml Clq was added to with 1×105 monocyte THP-1 for 24 h,then co-cultured with H.pylori(MOI 100:1)for 24 h.The protein was collected from the cells and extracted from the ice.The expression of the protein was detected by Western blot.The expression of LAIR-1,NF-?B p65,p-NF-?B p65,JNK,p-JNK,Akt,p-Akt,p38 MAPK,p-p38 MAPK.Results1.The effect of Clq on the expression of LAIR-1 on the surface of THP-1 cells infected with H.pylori:Compared with H.pylori infection group 2.61±0.16,2.64±0.07,2.14±0.17 and without H.pylori infection PBS group 0.95±0.17,0.64±0.19,0.52±0.03,the expression of LAIR-1 in THP-1 cells was significantly increased(P<0.05).H.pylori infection added C1q group was compared with H.pylori infection group.The expression of LAIR-1 was significantly decreased(P<0.05).2.The effect of Cl q on the secretion of IL-4 in THP-1 cells infected with H.pylori:Compared with H.pylori infection group(0.84±0.22)ng/L,(1.30±0.14)ng/L,(1.37±0.41)ng/L and without H.pylori infection PBS group(1.83±0.23)ng/L,(0.64±0.19)ng/L,(0.52±0.03)ng/L,the secretion of IL-4 was decreased.3.The effect of Clq on the secretion of IL-8 in THP-1 cells infected with H.pylori:Compared with H.pylori infection group(2.49±0.34)ng/L,(2.43±0.06)ng/L,(4.78±0.54)ng/L and without H.pylori infection PBS group(0.69±0.05)ng/L,(0.42±0.01)ng/L,(1.78±0.82)ng/L,and compared with H.pylori infection add C1q and H.pylori infection add PBS,the secretion of IL-8 was increased(P<0.05).4.The effect of Clq on the relative expression of IL-10 by THP-1 cells infected with H.pylori:Compared with H.pylori infection group(0.77±0.02)ng/L,(1.82±0.08)ng/L,(3.92±0.25)ng/L and without H.pylori infection PBS group(0.47±0.01)ng/L,(0.59±0.21)ng/L,(2.39±0.30)ng/L,and compared with H.pylori infection add Clq and H.pylori infection add PBS,the secretion of IL-10 was increased(P<0.05).5.The effect of Clq on the relative expression of Akt in THP-1 cells infected with H.pylori:Compared with H.pylori infection group 0.66±0.04,0.52±0.04 and without H.pylori infection group 0.44±0.04,0.36±0.03,the relative expression of activated Akt in THP-1 cells was significantly increased(P<0.05).6.The effect of C1q on the relative expression of p38 MAPK in H.pylori infected THP-1 cells:Compared with H.pylori infection group 1.05±0.07,0.87±0.05 and without H.pylori infection group 0.16±0.01,0.07±0.01,the relative expression of activated p38 MAPK in THP-1 cells was significantly increased(.P<0.05).Compared with C1q add H.pylori group 0.51±0.04 and without H.pylori group 0.73±0.05,and Compared with C1q add H.pylori group and PBS add H.pylori group,activated p38 MAPK was significantly decreased(P<0.05).7.The effect of Clq on the relative expression of JNK in H.pylori infected THP-1 cells:Compared with H.pylori infection group 0.73±0.03,0.30±0.02,0.24±0.04 and without H.pylori infection PBS group 0.15±0.01,0.18±0.01,0.08±0.01,and compared with H.pylori infection add C1q and H.pylori infection add PBS,activated JNK was increased(P<0.05).8.The effect of Clq on the relative expression of NF-?B p65 in H.pylori infected THP-1 cells:Compared with H.pylori infection group 0.93±0.32,1.21±0.41,1.34±0.12 and without H.pylori infection PBS group 0.13±0.11,0.21±0.12,0.19±0.16,activated NF-?B p65 was increased(P<0.05).Conclusion1.H.pylori infection may be expressed by increasing the expression of LAIR-1.It can increase the secretion of proinflammatory factor IL-8 and anti-inflammatory factor IL-10 by activating MAPK,JNK,Akt and NF-?B signaling pathways.Activation NF-?B signaling pathway inhibits secretion of anti-inflammatory factor IL-4.H.pylori may maintain persistent bacterial infection by balancing the secretion of inflammatory factors.2.When H.pylori infected,Clq affects the secretion of cytokines IL-8 and IL-10 by reducing the expression of LAIR-1 and inhibiting activation of MAPK signaling pathway. |
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