摘要目的 观察干细胞因子(SCF)在活化胰腺星状细胞(PSC)中的表达以及在PSC与肥大细胞黏附中的作用.方法 采用SD大鼠胰腺组织块培养分离活化PSC,应用RT-PCR免疫细胞化学染色法检测PSC的SCF mRNA和蛋白的表达;经不同剂量的抗SCF抗体中和PSC表面SCF,采用β-氨基已糖苷酶测定观察肥大细胞与PSC之间的黏附;PSC经10μg/ml丝裂霉素C处理后为饲养层细胞,与肥大细胞共培养,硫堇兰染色观察培养后2、3、5 d的肥大细胞生长情况.结果 活化的PSC表达SCFmRNA和蛋白.对照组、1mg/ml和10 mg/ml抗体处理组黏附的肥大细胞量分别为每视野(48±10)个、(28±8)个和(23±9)个,抗体组较对照组明显减少(P＜0.05).1 mg/ml和10 mg/ml抗体组的肥大细胞黏附率分别为(53±3)%和(45±5)%.PSC和肥大细胞在无血清条件下共培养1 d后,PSC有肥大细胞黏附;共培养3 d后黏附的数量增加;5 d后呈堆积状,有较多集落形成,PSC基本消失.结论 活化的PSC可合成分泌SCF,引起肥大细胞的黏附和生长.

abstractsObjective To investigate stem cell factor(SCF)expression in activated pancreatic stellate cell(PSC)and the effect of SCF on adhesion between PSC and mast cell(MC).Methods PSC of SD rat was separated and activated by pancreatic tissue culture;the expression of SCF mRNA and protein was detected bv RT-PCR and immunachemistry.SCF was blocked with different dose of anti-SCF antibody,the rate of adhesion between PSC and MC was determined by thiomn blue stain and β-hexasaminidase assay.PSC was stimulated by 10 μg/ml mitomycin-C and was co-cultured with MC,then the growth of MC was observed at 2,3,5 day,respectively.Results Activated PSC expressed SCF mRNA and protein,the number of adhered MC in the control,1mg/ml and 10 mg/ml antibody groups was 48±10,28±8,and 23±9,respectively;the difference between antibody and control group was statistically significant(P＜0.05).One day after PSC and MC was co-cultured without serum.there were MC adhesion;3 days later the rate of adhesion increased;5 days later the adhesion appeared bulk-like and lots of colonies were present,while PSC basically disappeared.Conclusions Activated PSC may synthesize,express SCF and induced adhesion and growth of MC.