| Ketamine in veterinary clinic often shows anesthetic,sedative,analgesic,antidepressant and hallucinogenic effects.The hallucinogenic effect is also called a dissociative narcotic because it dissociates the mind.Long-term use of ketamine can cause certain side effects,mainly in the nervous system and urinary system.Neurotoxicity studies are earlier and more extensive.At present,studies on urinary system are mainly focused on adult rats,and there are few studies on young animals.Ketamine is one of the most commonly used anesthetics for young animals in veterinary clinic because it has little effect on the respiratory system and fast anesthetic effect.Young animals are the peak of body development and the key period for the development of various organs.Ketamine administration during this period may cause certain renal effects in young animals,which may be related to cell burn death.Pyroptosis is a new form of programmed cell death based on the dual characteristics of cell necrosis and apoptosis.NLRP3/Caspase-1/GSDMD is the main pathway of cell pyrotosis.Clinically,inhibitors MCC950 and VX765 are often used to inhibit the occurrence of cell pyrotosis and have a protective effect on the kidney.Therefore,this study focus on the effects of ketamine on the kidney of young animals,to explore whether such effects are related to cell burn death,and to provide theoretical basis for clinical treatment.In this experiment,7-day-old SD rats were randomly divided into 6 groups: control group(group C),ketamine group(group K),ketamine+MCC950 inhibitor group(KM group),ketamine+VX765 inhibitor group(KV group),MCC950 group(M group),and VX765 group(V group).Group K is intraperitoneally injected with ketamine(20 mg/kg)once every 90 min,for a total of 5 times.KM group and KV group are intraperitoneally injected with MCC950 inhibitor(0.1mg/kg)and VX765 inhibitor(0.1 mg/kg)30 min before ketamine administration,and then the same procedure as K group.Group M and group V are only given corresponding inhibitors.Kidney tissue and blood are collected from young rats.The changes of serum BUN and CRE are detected by biochemical kit.The pathological structure of kidney is observed by HE staining.The m RNA expression levels of ASC,Caspase-1,GSDMD and NLRP3 related genes are detected by PT-PCR.The protein expressions of ASC,Caspase-1,Caspase-11,GSDMD and NLRP3 are detected by Western blot.The levels of IL-1β and IL-18 in kidney tissue and serum of young mice are detected by ELISA.The protein expression sites of scorpus-related proteins ASC,Caspase-1,GSDMD and NLRP3 were detected by immunohistochemistry.The test results are as follows:(1)Results of renal function test: the contents of BUN and CRE in 7-day-old young rats in group K are the highest,the index contents in KM group and KV group are significantly decreased compared with that in group K,and there are no significant change in group M,V and C.There are no significant change in all groups at 60 days of age.(2)Renal histological results: there are inflammatory cell infiltration in renal tubules and glomeruli in the K group by HE staining,and the skin cells in renal tubules are damaged and shed,and the glomeruli are accompanied by congestion.Compared with the K group,the renal tissue in the KM group and the KV group was significantly improved,with reduced inflammatory cell infiltration and repaired renal tubular dermal cells.The renal structure in the M group and the V group are basically the same as that in the C group,without abnormal pathological changes.(3)RT-PCR results:the m RNA expressions of ASC,Caspase-1,GSDMD and NLRP3 are significantly up-regulated in the kidney tissues of group K,and are significantly down-regulated in the KM and KV groups compared with the K group,while no significant differences are found in the M,V and C groups.(4)Western blot results: the protein expression levels of ASC,Caspase-1,Caspase-11,GSDMD and NLRP3 are the highest in group K,KM group and KV group are significantly down-regulated compared with that in group K,and there are no significant difference in group M,V and C.(5)ELISA results: the contents of IL-1β and IL-18 in serum and renal tissues of group K are the highest,and the contents of KM and KV groups are significantly decreased compared with that of group K,while there are no significant change among groups M,V and C.(6)Immunohistochemical results: The positive rate of related proteins are higher in group K,and the average optical density are significantly higher than that in group C.The positive rate of KM and KV group are lower,and the average optical density are decreased compared with that of K group.The results suggest that anaesthetizing young animals with ketamine for long periods of time can cause kidney damage.The degree of damage are significantly reduced after treatment with pyroptosis inhibitors,suggesting that ketamine-induced renal injury are related to pyroptosis.At the same time,giving a pyrotic inhibitor before anaesthesia in young animals have a protective effect on the kidneys. |
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