论文部分内容阅读
目的观察胰腺应激蛋白PSP/reg对胰腺星状细胞(PSC)合成和分泌基质金属蛋白酶(MMPs)及其组织抑制剂(TIMPs)以及RECK表达的影响。方法分离纯化慢性胰腺炎患者纤维化区的PSC,基因重组胰腺应激蛋白PSP/reg,以终浓度为10和100 ng/mL对PSC进行干预,实时荧光定量PCR检测MMP1/2、TIMP1/2及RECK基因表达,Western blot测定MMP1/2、TIMP1/2及RECK蛋白,细胞免疫荧光观察细胞膜表面RECK分布。结果 PSP/reg对MMP1/2、TIMP1/2及RECK表达无明显影响;PSP/reg轻度抑制PSC培养上清中MMP2水平(P<0.05),而显著抑制TIMP1/2水平(P<0.01);PSC细胞膜表面发现有RECK蛋白,PSP/reg减少PSC的RECK含量(P<0.01)。结论胰腺应激蛋白PSP/reg能够降低TIMPs:MMPs比率、减少RECK蛋白水平表达,从而解除对MMPs的部分抑制,使MMPs活性相对增高,有利于纤维化的分解消散,促进胰腺损伤后的再生修复。
Objective To investigate the effects of pancreatic stress protein PSP / reg on the synthesis and secretion of matrix metalloproteinases (MMPs), tissue inhibitors (TIMPs) and RECK in pancreatic stellate cells (PSCs). Methods PSC and PSNP were isolated and purified from fibrosis area of patients with chronic pancreatitis. PSCs were treated with 10 and 100 ng / mL final concentration of PSCs, and MMP1 / 2 and TIMP1 / 2 were detected by real-time fluorescence quantitative PCR And RECK gene expression. MMP1 / 2, TIMP1 / 2 and RECK protein were detected by Western blot. The distribution of RECK on cell membrane was observed by immunofluorescence. Results PSP / reg had no significant effect on MMP1 / 2, TIMP1 / 2 and RECK expression. PSP / reg slightly inhibited the level of MMP2 in the supernatant of PSC (P <0.05) RECK protein was found on the surface of PSC cell membrane, and PSP / reg decreased the RECK content of PSC (P <0.01). Conclusions Pancreatic stress protein PSP / reg can decrease the ratio of TIMPs to MMPs and decrease the expression of RECK protein, so that partial inhibition of MMPs can be lifted and the activity of MMPs can be relatively increased, which can facilitate the decomposition and dissipation of fibrosis and promote the regeneration and repair of pancreatic injury .