地氟醚对小鼠中枢及外周组织生物钟基因的影响

Effects of desflurane on central and peripheral clock genes in mice

目的探讨地氟醚对小鼠中枢及外周组织生物钟基因的影响及其作用机制。方法首先用不同浓度(3%、4%、6%、7%、9%)的地氟醚麻醉C57BL/6J小鼠,通过翻正反射确定地氟醚的有效作用浓度;然后分别将C57BL/6J小鼠和Per1^-/-/Per2^-/-小鼠分为对照组和地氟醚组,地氟醚组在小鼠饲养环境开灯后2 h用前述确定的地氟醚浓度(7%)处理,对照组则同时通入空气,2 h后采用RT-qPCR检测各组小鼠下丘脑、肾脏和肝脏组织时钟基因Per1、Per2、Dbp、Bmal1、Clock和Cry1的表达。结果①3%地氟醚处理后无小鼠达到翻正反射消失;4%、6%、7%、9%地氟醚处理后达到翻正反射消失的小鼠比例分别为16.7%、50%、100%和100%,确定地氟醚的作用浓度为7%。②与对照组比较,7%地氟醚麻醉C57BL/6J小鼠2 h后,下丘脑组织时钟基因Per2的表达量显著降低(P<0.01),其余5个时钟基因的表达量变化无统计学意义(P>0.05);肾脏组织中时钟基因Per1的表达量显著升高,而Bmal1、Dbp的表达量显著降低(P<0.05),另外3个时钟基因的表达量变化差异无统计学意义(P>0.05);肝脏组织时钟基因Per1的表达量显著升高(P<0.01),其余5个时钟基因的表达量变化差异无统计学意义(P>0.05)。与对照组比较,7%地氟醚麻醉Per1^-/-/Per^2-/-小鼠2 h后,下丘脑组织时钟基因Bmal1、Clock、Dbp的表达量显著降低(P<0.05);肝肾组织时钟基因Cry1、Clock的表达量显著降低(P<0.05),Bmal1的表达量变化差异无统计学意义(P>0.05)。结论地氟醚显著影响小鼠中枢及外周组织的生物钟相关基因,且存在不依赖Per基因的作用途径。

Objective To investigate the effect and mechanism of the anesthetic drug,desflurane,on central and peripheral clock genes in mice.MethodsC57BL/6J mice were anesthetized with different concentrations(3%,4%,6%,7%,9%)of desflurane,and then the effective concentration of desflurane was determined by detecting the loss of righting reflex.Then C57BL/6J mice and Per1^-/-/Per^-/-mice were respectively divided into air control group and desflurane treatment group.The mice from the latter group were given 7%desflurane treatment in 2 h after the light of feeding environment was turned on,and those of the control group was exposed to air at the same time.In 2 h later,the clock genes Per1,Per2,Dbp,Bmal1,Clock and Cry1 in the hypothalamus,kidney and liver tissues were detected by RT-qPCR.Results①Treatment of 3%desflurane resulted in no disappearance of righting reflex,while those of 4%,6%,7%and 9%caused the mice achieving loss of the righting reflex accounted for 16.7%,50%,100%and 100%.Therefore,the effective concentration of desflurane was determined as 7%.②Compared with the control group,treatment with 7%desflurane significantly decreased the expression of clock gene Per2 in the hypothalamus tissue in C57BL/6J mice(P<0.01),but had no effect on the expression of the other 5 clock genes(P>0.05);It also significantly increased the expression of the clock gene Per1,while significantly decreased the expression of the clock genes Bmal1 and Dbp in the kidney tissue(P<0.05),but there was no significant change in the expression of the other 3 clock genes(P>0.05);In the liver tissue,the expression of clock gene Per1 was significantly increased(P<0.01),while no changes were seen in the expression of the other 5 clock genes(P>0.05).Compare with the control group,the Per1^-/-/Per2^-/-mice had significantly decreased the expression of Bmal1,Clock and Dbp in the hypothalamus tissue after treatment with 7%desflurane(P<0.05);obviously reduced expression of Cry1 and Clock(P<0.05),but no notable change in that of Bmal1(P>0.05)in the liver and kidney tissue.ConclusionDesflurane significantly affects the circadian clock genes in mouse central and peripheral tissues,and there is a pathway independent form Per gene.