Injury Of Bladder Tissue In Rats Induced By Ketamine Abuse And Its Mechanism Of Shuangan Oral Liquid

Objective: In this study,SD male rats were used as the research object to establish an animal model of bladder tissue damage caused by ketamine abuse,and on this basis,withdrawal and shuangan oral liquid were used for treatment.To explore the functional status of bladder epithelium and the change of apoptosis in SD rats,in order to clarify the damage of bladder tissue caused by ketamine abuse and the treatment mechanism of shuangan oral liquid,to provide new treatments for clinical treatment and to provide new drug developmentexperimental basis.Methods:1.the 96 only 10-week-old SD male rats were randomly divided into 8 groups,respectively as a control group and a ketamine modeling group [3 groups,each given 10 mg/kg of ketamine(K10 group),30 mg/kg(K30 group)and 60 mg/kg)],ketamine 30 mg/kg withdrawal treatment group(K30T group)and ketamine 30 mg/kg shuangan oral liquid treatment group [3 groups,shuangan oral liquid 3750mg/kg(K30L group),11250mg/kg(K30M group)and 18750mg/kg(K30H group)],12 in each group.After 30 days of continuous intraperitoneal injection with different doses of ketamine/normal saline in each group of rats,the K30 T group was discontinued for 2 weeks,and the shuangan oral liquid treatment group was given gavage of low,medium and high doses of shuangan oral liquid for 2 weeks.Rats in each group were sacrificed 24 hours after the last administration.2.observe the behavioral changes of rats in each group during ketamine administration.3.observe the urinary system(kidney,ureter,bladder)tissues of rats with HE staining.Changes in structure and organ coefficients.4.realtime-PCR,western blot,and immunohistochemistry were used to detect(1)tight junction proteins E-cad and ZO-1 of bladder epithelial cells;(2)histamine receptor H1 of bladder tissue;(3)expression of endoplasmic reticulum stress pathway members GRP78,PERK,ATF-4,CHOP and caspase-12 in bladder epithelial cells.Results: Behavioral observation: compared with the control group,the rats in the model group had significantly enhanced mobility after administration.With the increase of drug dose,sneezing,shaking head,unstable standing,falling and aggressive behaviors appeared successively,and the time of drug effect was prolonged with the increase of drug dose.Weight change: There was no statistical difference between the control group and the model group(P>0.05).The weight gain was found in the K30 T and K30 L groups,and the K30 M and K30 H groups were increased after shuangan oral liquid treatment or withdrawal treatment.The body weight of the group decreased,and the weight gain of the K30 H group was significantly lower than that of the other three groups(P<0.01).The results of changes in the urinary structure of the rats showed that compared with the control group,the structure of the urinary system(kidney,ureter,bladder)in the model group changed to varying degrees compared with the control group.The degree of deepening was increased,and the shuangan oral liquid treatment group and the withdrawal treatment group could effectively improve the degree of tissue lesions.The results of organ coefficient determination in the animal model showed that there was no statistical difference in the organ coefficient of kidney,bladder and ureter in each group(P>0.05),indicating that the kidney,ureter and bladder tissues of each group had not yet produced substantial lesions.The expression of tight junction protein between cells of rat bladder epithelial tissue showed that compared with the control group,different doses of ketamine could inhibit the expression of E-cad and ZO-1,the withdrawal treatment group did not change significantly and while the different doses of shuangan oral liquid could all its expression was upregulated,and the K30 M group had the best effect,suggesting that ketamine may reduce the expression of tight junction protein in bladder epithelial cells,resulting in impaired bladder epithelial tissue barrier function.The improvement effect of withdrawal treatment group is not good and Shuangan oral liquid can effectively improve this situation and play a role in repairing the function of bladder epithelium.The expression of histamine receptors in the bladder tissue of rats showed that H1 in the bladder tissue of the model group showed high expression,while its expression was down-regulated by ketamine withdrawal treatment and shuangan oral liquid treatment.It is suggested that ketamine can increase the permeability of bladder epithelium by up-regulating the expression of histamine receptors in bladder tissue,and shuangan oral liquid treatment group and withdrawal group can improve this trend.Detection of the apoptosis signal pathway induced by endoplasmic reticulum stress in rat bladder epithelial cells shows that the different doses of ketamine can simultaneously upregulate the protein expression of GRP78,PERK,ATF-4,CHOP,and Caspase-12,making bladder tissue being in the endoplasmic reticulum stress state and triggering apoptosis;the withdrawal treatment group and shuangan oral liquid treatment group can reverse this trend,and the repair effect of shuangan oral liquid is better than the withdrawal treatment group.It is suggested that ketamine can induce apoptosis through the endoplasmic reticulum stress pathway.Both withdrawal treatment and shuangan oral liquid can effectively inhibit this effect.Conclusion: Intraperitoneal injection of ketamine can successfully establish an animal model of bladder tissue damage caused by ketamine abuse.Ketamine can cause rat bladder tissue damage by causing changes in the urinary tissue structure of rats,reducing the expression of tight junction protein in rat bladder epithelial cells,up-regulating the expression of bladder histamine receptor H1,and triggering rat bladder epithelial cell apoptosis.Both withdrawal treatment and shuangan oral liquid can effectively improve bladder tissue damage,and shuangan oral liquid has better repair effect.