感染后膈肌无力分子机制的研究策略  

Mechanism of and strategy to study the diaphragmatic weakness in sepsis

作　　者：姬新颖[1] 李涛[1] 张军[1] 刘广超[1] 李淑莲[1] 刘峰涛[1] 牛保华[1] 黄红莹[1] 白慧玲[1] 蒋杞英[1] 马远方[1] 

机构地区：[1]河南大学医学院河南省细胞与分子免疫学医学重点实验室,河南开封475004

出　　处：《河南大学学报（医学版）》2012年第3期167-170,共4页Journal of Henan University:Medical Science

基　　金：国家自然科学基金项目(81071327;81270142);河南省科技厅重点科技攻关项目(11430051026);河南大学省部共建项目(SBGJ090712)

摘　　要：感染是膈肌无力的重要原因之一,但对其认识匮乏,分子机制更鲜有研究。有实验表明内毒素处理小鼠后膈肌中活性PKR和上游MAPK激活剂MEK3/6显著增加,而这种变化的方式与Caspase转导通路的激活一致。通过建立小鼠膈肌和肌细胞感染模型,采用Western印迹、酶活性测定、流式细胞检测和膈肌肌力测量等方法,探讨感染致膈肌肌力下降与Caspase、p38等分子的关系;运用化学抑制剂和基因抑制剂进一步确认其在肌肉功能障碍中的作用。p38与细胞生长有关,直接用siRNA沉默后肌细胞无法分化成熟,影响进一步研究。为此科学家培育出新一代p38转基因小鼠,将终止密码、Cre-Loxp与负显性基因(DN)有机结合,定时定点控制肌细胞DN基因表达,克服RNAi之不足,开创肌细胞模型中重要基因功能研究的新路径。可以预期,通过证实感染造成的膈肌细胞功能障碍与p38通路之间的关系,人们能够将从分子水平探讨感染引起膈肌肌力下降的机理和可能的防治策略。Sepsis is one of the key factors in diaphragmatic weakness.Unfortunately the phenomenon was less concerned by physicians and scientists,not to say the molecular mechanism.Some study showed that there were a obvious increase of active PKR and its upstream stimulator MEK3/6 in the diaphragm of mice exposed to endotoxin,which was similar to the activation of caspase signaling.The possible reliable methods that can be used to the study of diaphragmatic weakness are： 1） the relationship of diaphragmatic weakness with some molecules such as p38,JNK1,PKR,TIPE2,PCNP can be confirmed by Western blotting,enzyme activity measurement,flow cytometry and detection of force generation of diaphragm.2）then the chemical and genetic inhibitors can be administrated to demonstrate the roles of the above-mentioned molecules in diaphragmatic weakness after sepsis.p38,significantly related to cell growth,can not be silenced by siRNA because of its vital effect on cellular differentiation and the following maturation after silencing.To overcome the shortcoming of RNAi,our group developed a novel transgenic-knockout mouse that combines the features of Cre-Loxp system with dominant-negative p38（p38-DN） gene after stop signal of transcription（four poly A sequences） which inhibits the normal expression of p38.This new method can control the p38-DN expression by tamoxifen and provide a different pathway to knockout gene.It is anticipated that we found the possible mechanism of diaphragmatic weakness after infection and the strategy to prevent and cure it at molecular level.This review tried to gather the recent progress in the relationship of p38,JNK1 and PKR with diaphragmatic weakness.

关 键 词：感染 膈肌无力 分子机制 P38 CASPASE 

分 类 号：R332.1[医药卫生—人体生理学]