摘要
将分离的仔猪胃粘膜细胞分为4组,在含10%胎牛血清的DM EM高糖培养液中培养24 h后,分别在新鲜的基础培养液中添加0(对照组)、0.001(低水平组)、0.01(中水平组)和0.1(高水平组)g/L的半胱胺盐酸盐,继续培养20 h后收集细胞,测定细胞H+-K+-ATPase活性,同时用相对定量RT-PCR法测定细胞H+-K+-ATPase和生长抑素(SS)mRNA的相对丰度。结果表明:(1)低水平的半胱胺盐酸盐对H+-K+-ATPase的表达和活性没有影响(P>0.05),但中、高水平的半胱胺盐酸盐显著提高了H+-K+-ATPase的表达和活性(P<0.05),其中H+-K+-ATPase mRNA的相对丰度分别提高了36%和44%,H+-K+-ATPase活性分别提高了57%和53%。(2)低水平半胱胺盐酸盐对SS mRNA的表达没有影响(P>0.05),中、高水平的半胱胺盐酸盐显著提高了SS mRNA的相对丰度(P<0.05),提高幅度均为52%。以上结果提示半胱胺盐酸盐可增强体外培养的仔猪胃黏膜细胞H+-K+-ATPase的表达及活性,而这一作用可能由SS所介导。
Mucosal cells isolated from gastric tissue of piglets were divided into 4 groups and then cultured in DMEM medium （high glucose） supplemented with 10% fetal calf serum. After 24 hours ,cysteamine hydrochloride was added into freshly replaced medium at the concentration of 0, 0. 001,0. 01,and 0. 1 g/L for the control,low,moderate and high concentration groups,respectively. After 20 hours of treatment ,cells were collected and the H^＋-K^＋-ATPase activity was analyzed. The relative abundances of somatostatin and H^＋-K^＋-ATPase mRNA were also determined by semi-quantitative RT-PCR. The results showed that ： （1）Low concentration of cysteamine hydrochloride exhibited no effect on both mRNA expression and activity of H^＋-K^＋-ATPase,while moderate and high concentrations of cysteamine hydrochloride markedly increased both H^＋-K^＋-ATPase mRNA expression by 36% and 44% ,and H^＋-K^＋-ATPase activity by 57% and 54% ,respectively; （2）Low concentration of cysteamine hydrochloride had no influence on SS mRNA expression,whereas the moderate and high concentrations of cysteamine ） hydrochloride significantly increased SS mRNA expression by 52%. The results suggest that cysteamine hydrochloride improve both mRNA expression and activity of H^＋-K^＋-ATPase in gastric mucosal cells of piglets in vitro,and this effect may be mediated by somatostatin.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2006年第4期411-414,共4页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(30270975)
