Effects Of Desflurane And Propofol On The Central And Peripheral Clock Genes Of Mice

Objectives:1.To investigate the effect of gas general anesthetic drug desflurane on central and peripheral clock of mice,and its mechanism of action;2.To investigate the effect of intravenous general anesthetic drug propofol on the rhythm of the clock genes.Methods:1.C57BL/6J?C57?mice were anesthetized with different concentrations?3%,4%,6%,7%,9%?of desflurane.The effective concentration of desflurane was determined by detecting whether or not the righting reflex disappeared.2.C57 mice were divided into two groups:air control group and desflurane treatment group.Each experimental groups were treated with the identified desflurane concentration?7%?at ZT2,and the control group was exposed to air at the same time.After 2 hours,the clock genes Per1,Per2,Dbp,Bmal1,Clock and Cry1 in the hypothalamus,kidney and liver tissues were detected by qPCR.3.Extract the DNA of the mouse to be identified and perform PCR identification.The product was detected by agarose gel electrophoresis.4.Per1^-/-/Per2^-/-mice were divided into two groups:air control group and desflurane treatment group.Each experimental groups were treated with the identified desflurane concentration?7%?at ZT2,and the control group was exposed to air at the same time.After 2hours,the clock genes Dbp,Bmal1,Clock and Cry1 in the hypothalamus,kidney and liver tissues were detected by qPCR.5.Replace the medium of U2OS cells with XM medium.According to the final concentration of propofol?1.5?M?6?M?12?M?50?M?,add the corresponding volume of propofol to each dish,and add the PBS solution to the control group.Record in the Lumicycle photometer.Results:1.After treatment with 3%desflurane,no mice reached the righting reflex disappeared;after 4%,6%,7%,9%desflurane treatment,the proportion of mice that achieved LORR were16.7%,50%,100%and 100%.Therefore,the effective concentration of desflurane is determined to be 7%.2.In C57BL/6J mice treated with 7%desflurane,compared with control group,the expression of clock gene Per2 in hypothalamus is significantly decreased?P<0.05?,while there was no significant change in the expression of the other five clock genes?P>0.05?.In kidney,the expression of the clock gene Per1 is significantly increased,the expression of the clock gene Bmal1 and Dbp significantly decreased?P<0.05?,but there was no significant change in the expression of the other three clock genes?P>0.05?.In liver,the expression of clock gene Per1 is significantly increased?P<0.05?,but there was no significant change in the expression of the other five clock genes?P>0.05?.3.The 440bp band refers to Per1^-/-,and the 320bp band refers to Per2^-/-,thus samples that show bands at both 320bp and 440bp are Per1^-/-/Per2^-/-.The 416bp band refers to Per1^+/+,and the 388bp band refers to Per2^+/+,thus samples that show bands at both 416bp and 388bp are Per1^+/+/Per2^+/+.4.In Per1^-/-/Per2^-/-mice treated with 7%desflurane,compared with control group,the expression of Bmal1,Clock and Dbp is significantly decreased in hypothalamus?P<0.05?;the expression of Cry1 and Clock is significantly decreased in liver and kidney?P<0.05?,but there was no significant change in the expression of Bmal1 and Dbp?P>0.05?.5.Compare with the control group,1.5?M propofol caused an increase in the amplitude of the Per2,after the concentration of propofol treatment increased,the amplitude of the Per2decreased,and the degree of decrease was concentration-dependent,while the period of Per2fluctuated between 22-23 hours in control group and propofol group,there have no significant change?P>0.05?.Conclusions:1.Gas anesthetic desflurane significantly affects the circadian clock of central and peripheral tissues,but does not directly through the Per gene.2.Intravenous anesthetic propofol reduces the amplitude of clock gene Per2 in cells.