IL-18对胰腺星状细胞及其趋化因子CX3CL1表达的影响
The influence of IL-18 on pancreatic stellate cells and CX3CL1 expression
目的 观察IL-18对胰腺星状细胞E-钙黏素(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)及趋化因子CX3CL1表达水平的影响.方法 人胰腺星状细胞(PSCs)株HPaSteC常规培养、传代,应用5、25、50、100 μg/L IL-18干预72 h,以未干预细胞为对照组.收集各组细胞,采用RT-PCR和蛋白质印迹法检测细胞E-cadherin、α-SMA、CX3CL1 mRNA及蛋白表达量.结果 对照组及5、25、50、100 μg/LIL-18干预组PSCs的E-cadherin mRNA表达量分别为1.03±0.17、0.77±0.15、0.89±0.12、0.54±0.11、0.46 ±0.06;α-SMA mRNA表达量为1.03±0.19、0.85±0.14、1.33±0.22、1.60±0.14、1.94±0.09;CX3CL1 mRNA表达量为1.01 ±0.08、0.88 ±0.25、0.86±0.17、1.58 ±0.26、1.83±0.13.干预组E-cadherin mRNA的表达下调,α-SMA及CX3 CL1 mRNA表达上调,其中100 μg/L干预组与对照组的差异均有统计学意义(P <0.05或<0.01).对照组及5、25、50、100 μg/L IL-18干预组PSCs的E-cadherin蛋白表达量分别为1.00 ±0.14、1.14 ±0.04、1.14 ±0.07、0.85 ±0.08、0.80±0.06;α-SMA蛋白表达量为1.00 ±0.02、0.77±0.07、1.29±0.02、1.59 ±0.07、1.70±0.02;CX3CL1蛋白表达量分别为1.00±0.05、1.03 ±0.05、1.37 ±0.06、1.46 ±0.18、1.45 ±0.12.干预组E-cadherin蛋白表达下调,但各组间的差异无统计学意义;α-SMA蛋白表达上调,25、50、100 μg/L IL-18干预组与对照组的差异有统计学意义(P值均<0.01);CX3CL1蛋白表达上调,其中100μg/L干预组与对照组的差异有统计学意义(P<0.05).结论 IL-18能激活人PSCs并上调PSCs趋化因子CX3CL1的表达.
更多Objective To evaluate the effect of IL-18 on the expression of E-cadherin,α-SMA and CX3 CL1 in pancreatic stellate cells (PSCs).Methods The human PSC line HPaSteC was routinely cultured and passaged.Five,25,50 and 100 μg/L IL-18 was used to treat PSCs for 72 h and the untreated PSCs were used as control.Treated and untreated cells were both collected,and RT-PCR and Western blot were used to detect mRNA and protein expression of E-cadherin,α-SMA and CX3CL1 respectively.Results The mRNA expressions of E-cadherin in control group and 5,25,50 and 100 μg/L IL-18 treated group were 1.03 ± 0.17,0.77 ±0.15,0.89 ± 0.12,0.54 ± 0.11 and 0.46 ± 0.06.The mRNA expression of α-SMA were 1.03 ± 0.19,0.85 ± 0.14,1.33 ± 0.22,1.60 ± 0.14 and 1.94 ± 0.09;The mRNA expression of CX3CL1 were 1.01 ±0.08,0.88 ±0.25,0.86 ±0.17,1.58 ±0.26 and 1.83 ±0.13.The mRNA expression of E-cadherin in IL-18 treated group were down-regulated,while the mRNA expression of α-SMA and CX3CL1 were up-regulated,and the differences between control and IL-18 100 μg/L treated group were statistically significant (P < 0.05 or < 0.01).The protein expression of E-cadherin in control group and 5,25,50 and 100 μg/L IL-18 treated group were 1.00 ±0.14,1.14 ±0.04,1.14 ±0.07,0.85 ±0.08 and 0.80 ±0.06.The protein expression of α-SMA were 1.00 ± 0.02,0.77 ± 0.07,1.29 ± 0.02,1.59 ± 0.07 and 1.70 ± 0.02;The protein expression of CX3CL1 were 1.00 ± 0.05,1.03 ± 0.05,1.37 ± 0.06,1.46 ± 0.18 and 1.45 ± 0.12.The protein expression of E-cadherin was down-regulated but no significant differences were observed among different groups.The protein expression of α-SMA was up-regulated and the differences between control group and 25,50 and 100 μg/L IL-18 treated groups were statistically significant (all P <0.01).The protein expression of CX3CL1 was up-regulated and the differences between control group and 100 ng/ml IL-18 treated group were statistically significant (P <0.05).Conclusions IL-18 can activate PSCs and up-regulate the expression of chemokine CX3CL1.
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