Nesfatin-1对离体胃黏膜细胞胃酸分泌的影响  

Nesfatin-1 inhibits gastric acid secretion by cultured rat gastric mucosa cells

作　　者：李静慧[1] 李学良[1] 吴静[1] 贾方圆[1] 林琳[1] 

机构地区：[1]南京医科大学第一附属人民医院消化科,江苏省南京市210029

出　　处：《世界华人消化杂志》2012年第13期1123-1130,共8页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.81070308/H0308~~

摘　　要：目的:研究nesfatin-1对离体培养的大鼠胃黏膜酸分泌的影响,探讨nesfatin-1对H+/K+-ATP酶mRNA及蛋白表达的影响.方法:酶解法分离大鼠胃黏膜细胞,细胞免疫荧光检测法鉴定细胞.用不同浓度的nesfatin-1(10-4-10-1μmol/L)对胃黏膜细胞进行处理0、1、2、3、4h,设立空白对照组,以14C-氨基比林摄取为酸分泌指标,检测nesfatin-1对大鼠离体的胃黏膜细胞酸分泌的影响.用RT-PCR法及Western印迹法检测nesfatin-1对胃黏膜细胞H+/K+-ATP酶alpha(α)亚基和beta(β)亚基mRNA及蛋白表达的影响.结果:Nesfatin-1在10-1μmol/L浓度下,在1、2h能够抑制离体培养的大鼠胃黏膜细胞的酸分泌.Nesfatin-1(10-1μmol/L)在1、2、3h均能够抑制H+/K+-ATP酶α亚基mRNA表达水平;在1、2h能够抑制H+/K+-ATP酶β亚基mRNA表达水平,分别与对照组相比,差异有统计学意义(均P<0.01).Nesfatin-1(10-4-10-1μmol/L)作用胃黏膜细胞2h时,呈剂量依赖性抑制α亚基和β亚基mRNA表达水平,分别与对照组相比,差异有统计学意义(均P<0.01).Nesfatin-1(10-1μmol/L)在1、2、3h能够抑制α亚基蛋白表达水平;在2、3h能够抑制β亚基蛋白表达水平,分别与对照组相比,差异有统计学意义(均P<0.01).Nesfatin-1(10-3-10-1μmol/L)作用胃黏膜细胞2h时,呈剂量依赖性抑制α亚基和β亚基蛋白表达水平,与对照组相比,差异有统计学意义(均P<0.01).结论:Nesfatin-1能抑制离体培养的大鼠胃黏膜细胞的酸分泌,有可能是通过下调H+/K+-ATP酶α亚基和β亚基的mRNA及蛋白表达的水平影响酸分泌.AIM：To clarify the effect of nesfatin-1 on gastric acid secretion and the expression of the H＋/K＋ATPase mRNA and protein in rat gastric mucosa cells in vitro.METHODS：Gastric mucosa cells were isolated from SD rats by enzymolysis and identified by immunofluorescence staining.Cultured rat gastric mucosa cells were divided into control group and nesfatin-1 group,and the nesfatin-1 group was pretreated with different concentrations（0,10-4,10-3,10-2,10-1 μmol/L） of nesfatin-1 for different durations（0,1,2,3,4 h）.The effect of nesfatin-1 on gastric acid secretion was investigated by monitoring 14C-aminopyrine（14C-AP） accumulation,and the expression of H＋/K＋ATPase α and β subunit mRNA and protein wasexamined by real-time PCR and Western blot.RESULTS：Pretreatment with nesfatin-1 at a dose of 10-1 or 10-2 μmol/L for 2 or 3 h inhibited gastric acid secretion,but nesfatin-1 at a dose of 10-3 or 10-4 μmol/L had no such effect.Nesfatin-1 at a dose of 10-1 μmol/L inhibited the expression of H＋/K＋-ATPase α subunit mRNA after pretreatment for 1,2,or 3 h and inhibited the expression of H＋/K＋-ATPase β subunit mRNAs after pretreatment for 1 or 2 h.In the dose range between 10-4 to 10-1 μmol/L,nesfatin-1 dose-dependently inhibited the expression of H＋/K＋-ATPase α subunit and β subunit mRNA after pretreatment for 2 h.Nesfatin-1 at a dose of 10-1 μmmol/L inhibited H＋/K＋-ATPase α subunit protein expression after pretreatment for 1,2 or 3 h and inhibited H＋/K＋-ATPase β subunit protein expression after pretreatment for 2 or 3 h.In the dose range between 10-3 to 10-1 μmol/L,nesfatin-1 dosedependently inhibited H＋/K＋-ATPase α and β subunit protein expression after pretreatment for 2 h.CONCLUSION：Our data suggest that nesfatin-1 inhibits gastric acid secretion by rat gastric mucosa cells in vitro possibly by down-regulating the expression of H＋/K＋-ATPase mRNA and protein.

关 键 词：NESFATIN-1 胃酸 胃黏膜细胞 H+K+-ATP酶 

分 类 号：R363[医药卫生—病理学]