Effects Of Propofol And Midazolam On Kidney And Lung Apoptosis Following Ischemia/reperfusion Of Hind Limbs In Rats

Objective In this study,we investigated the pathogenesis of injury of lung and kidney following ischemia/reperfusion of hind limbs in rats.The effects of anti-apoptosis of propofol and midazolam on the injury of the organs were studied using TUNEL and immunohistochemistry methods to observe the changes in cell apoptosis,expression of Bcl-2 and caspase-3 in lung and kidney tissue,serum creatinine and urea nitrogen.Methods Thirty-six healthy male SD rats weighing 220-250g were anesthetized with 10% urethane (1.0g·kg -1) and then randomly assigned into six groups.①sham group ( n=6),the rats received the operation without other treatment.②I1h group,ischemia in hind limb for 1hour and followed by 4 hours of reperfusion (n=6).③I3h group,ischemia in hind limb for 3hours and followed by 4 hours of reperfusion (n=6).④Propofol treated group(Pro group, n=6),before ischemia the rats were received a bolus of 0.5% propofol 5mg·kg-1,then continuously were infused as10 mg·kg-1·h-1 until the end of experiment.⑤Midazolam treated group(Mizo group, n=6),the rats were received a bolus of 0.5mg·kg-1iv,and were infused as 0.2mg·kg-1·h-1.⑥10% intralipid treated group(Intr group, n=6).The animals in other groups were administered with an equivalent dose of 0.5% intralipid in Intr group or normal saline in I1h,I3hgroup.At the end of the experiment,all rats were killed by carotid bloodletting,the blood was centrifuged,the lung and kidney tissue were sampled to observe the apoptosis,determine the expression of Bcl-2,caspase-3,the changes of DNA.Results①The variety of serum creatinine,urea nitrogen in all groups had normal value.②The TUNEL results: There were no significant apoptosis changes in sham group. Ischemia/reperfusion significant increased apoptosis index of lung and kidney.While propofol and midazolam significantly attenuated these changes induced by ischemia/reperfusion of hind limbs as compared with group of Intr and group of I3h.③The expression of Bcl-2, caspase-3: Compared with the sham group, the expression of Bcl-2, caspase-3 in group I1h and I3h increased obviously.Propofol and midazolam could attenuate this increase in caspase-3 but enhance the increase in Bcl-2.The expression of caspase-3 positively correlated to apoptosis.④There was no DNA fragment in group sham,but the rest groups acquired various DNA fragment.Conclusions①Apoptosis of kidney and lung participates in the injury induced by ischemia/reperfusion of hind limbs.②Propofol and midazolam have inhibitive effect on lung, kidney apoptosis following ischemia/reperfusion of hind limbs in rat.They may inhibit the expression of promoting apoptosis genes and up-regulate the anti-apoptosis genes, which might imply that propofol and midazolom have protective effect by relieving injury.