| Objective: To investigate the effect of MCP-1/CCR2 signal pathway on pathogenesis of interstitial cystitis.To study the therapeutic effect of CCR2 inhibitor and sodium cromoglycate in interstitial cystitis mice model.Methods: In this study,SV-HUC-1 and HMC-1 were cultured to explore the interaction between MCP-1 and mast cells.Transwell assay was used to analyze the chemotaxis of MCP-1 on mast cells.different concentrations of LPS respectively cocultured with SV-HUC-1 and HMC-1.The level of MCP-1 and histamine in supernatant was measured by ELISA method different concentrations of MCP-1 cocultured with HMC-1,the level of histamine in HMC-1 supernatant was measured by ELISA method.we establish a mouse model of interstitial cystitis with PS and LPS,then IC mice were injected intraperitoneally with sodium cromoglycate and CCR2 inhibitor,respectively,while we established the control group.the levels of histamine and MCP-1 in urine were measured by ELISA method.bladder tissue are collected after euthanizing the animals.HE staining and toluidine blue staining were used to observe the inflammation of bladder tissue and the number,distribution of the mast cells of bladder tissue.Result: 1.Transwell experiments showed that the chemotactic ability of the experimental group was significantly higher than the control group,and the chemotactic ability of the experimental group was positively correlated with the concentration of MCP-1;2.The level of MCP-1 secreted by SV-HUC-1 was significantly correlated with LPS concentration and stimulation time.The peak level of MCP-1 was achieved after 24 h exposure to 50ng/m L of LPS.When mast cells stimulated by different concentrations of LPS,the peak level of MCP-1 secretion was achieved after 48 h exposure to 50ng/m L of LPS.the level of MCP-1 secreted by HMC-1 was significantly correlated with LPS concentration,but it was not statistically significant.with the stimulation time.Mast cells stimulated by LPS released large amounts of histamine.Secretion of histamine stimulated by LPS in different concentrations(10,50,100 ng/m L)showed significant differences with normal group.Histamine reached its peak level after mast cells exposed to LPS(100 ng/m L)for 48 h,and the level of histamine showed significant correlation with LPS concentration and stimulation time.3.HMC-1 was exposed to different concentrations of MCP-1,and ELISA method was used to measure the the level of histamine at 12 h,24h and 48 h.The level of histamine increased with the concentration of MCP-1 when MCP-1 stimulation to mast cells.The level of histamine increased with the stimulation time,and showed significant diffenences in the 10 ng / m L,50 ng / m L and 100 ng / m L groups.The results showed that secretion of histamine was correlated with time and dose of MCP-1(p<0.01).4.The histopathology of bladder tissue in IC mice group showed obvious inflammation and partly exfoliation of the bladder epithelium.Increased of mast cells infiltration was seen in bladder epithelial submucosal and detrusor.Bladder inflammatory response and exfoliation of urinary tract epithelial cells significantly reduced and infiltration of mast cells decreased obviously in the IC mice model group treated by CCR2 inhibitor and cromoglycine respectively compared with contorl group.5.The levels of MCP-1 and histamine in the urine of IC mice group were higher than those of the normal mice group.The levels of histamine and MCP-1 in the urine of mice treated with CCR2 inhibitor and cromoglycans were lower than those of IC miceConclusion: 1.MCP-1 / CCR2 signal axis play a significant role through the mast cells in the pathogenesis of IC;2.The IC mice bladder inflammation.can be effectively improved by blocking the MCP-1 / CCR2 signal axis and treating with the Sodium cromoglycate that was able to stabilize mast cell membrane. |
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