鼠肥胖抑制素RIA的建立及初步应用

Establishment of a Method of Murine Obestatin RIA and its Primary Application

目的:研究鼠肥胖抑制素(Obestatin)RIA的建立方法,并探讨Obestatin在创伤应激反应中的作用。方法:利用人工合成的鼠Obestatin免疫新西兰白兔制备抗血清,采用氯胺-T法碘标记Obestatin抗原,从而制备Obestatin RIA。建立小鼠盲肠结扎穿孔致脓毒症模型,设立假手术和损伤后3h、6h、9h组,每组9只,并采用自制Obestatin RIA检测损伤后各组血清Obestatin的水平。结果:所用Obestatin RIA的标准曲线形态和测量结果均良好。与假手术组血清Obestatin水平相比,损伤后6 h组显著升高,而损伤后3h、9h组与之无显著差异。同时,损伤后6h组还有显著高于损伤后3h、9h组的趋势。结论:所建立的鼠Obestatin RIA灵敏度高、简便、可靠,能够用于大鼠、小鼠相关样品的检测。Obestatin可能作为一种创伤应激因子参与脓毒症后内环境紊乱的调控。

Objective To develop a method of murine obestatin RIA and study the role obestatin played in traumatic stress responses.Methods New Zealand white rabbits were immunized with synthetic murine obestatin to obtain anti-serum,while chloramines-T method was used to iodinate obestatin antigen and a method of RIA for obestatin was established.A mouse sepsis model made with cecal ligation and puncture was established.Serum obestatin levels in the sacrificed models were determined with this method of RIA at 3h,6h,9h after the cecum puncture respectively（5 animals each time） as well as in 5 animals after sham operation.Results Both the shape of standard curve and metrical results of the obestatin RIA were satisfactory.Serum obstatin levels at 6h after injury were significantly higher than those in animals with sham operation out the levels at 3h and 9h were not much different from those in sham operation animals.Meanwhile,the levels at 6h after injury expressed a trend to be significantly higher than that at 3h or 9h after injury.Conclusion The established method for murine obestatin RIA is highly sensitive,simple and reliable,and it can be used to detect samples from rats and mice.Obestatin may be a traumatic stress factor participating in the modulation of homeostasis after sepsis.