Protective Effect Of AH₂QDS On Kidney Poisoned By Paraquat

Objective:Through experiments,the protective mechanism of AH2QDS on paraquat poisoned kidney was explored from the aspects of oxidative stress,inflammatory response and apoptosis,And the role of the Apelin/APJ system in detoxification,providing theoretical basis for further improving the treatment rate of AH2QDS.Methods:(1)72 SD rats were randomly divided into Control,PQ,PQ+Sivelestat and PQ+AH2QDS groups with 18 rats in each group.Sivelestat has stable anti-inflammatory effects and served as a positive control group.20%PQ 200mg/kg(pesticide,LD50=200 mg/kg)was given intragastric administration to establish poisoning model.Sivelestat intraperitoneal injection and AH2QDS intragastric intervention were administered 2 hours after poisoning.Samples were taken on days 1,3 and 7 for detection.(2)Outcome measures:general condition,survival rate and body weight change of rats;PQ content in digestive tract and kidney tissue;Renal morphology and HE staining of renal tissue;The expression of Apelin in renal tissues was determined by immunofluorescence technique.The Levels of IL-6 and TNF-α inflammatory factors and the protein expressions of Apelin/APJ,NF-κBp65,Caspase 1,Caspase 8,GRP78 and CHOP were detected by Western blot.(3)In order to understand the direct therapeutic effect of AH2QDS,IC50 of PQ(pure product)and the effect of Sivelestat and AH2QDS on cell viability were detected by CCK8 assay in human renal tubular epithelial cells.(4)The toxicity model was established according to IC50 of PQ intervention for 24 hours,remove the original culture medium,and then the cells were treated with Sivelestat and AH2QDS with no effect on cell viability for 24 hours.Cell viability was detected by CCK8,the content of OH·was measured by colorimetry,the expression of IL-6 and TNF-α were detected by Elisa,and the direct effect of AH2QDS on cell therapy was observed.Results:(1)The rats in PQ group were poisoned 1hour～2 hours Later,they showed poor mental performance,Less movement,squinting,no eating,chills,vertical hair,hair become yellow,ball Like rolling,dark green thin stools,poor stimulus response,staggered walking and unstable standing.On the second to third days,most of the above symptoms were aggravated,and some of the heavier rats showed blood scabs at the nose and mouth,obvious emaciation,the spirit of dull,asthma,aspirated Laryngeal vocalization,cyanosis of Limb sand the whole body;Fidgeting,convulsions,death within 1 to 2 days.PQ+AH2QDS group and PQ+Sivelestat group showed relatively mild symptoms.Most of the rats in PQ+AH2QDS group did not affect eating after poisoning,and the symptoms were obviously mild,average mental activity,and the weight gain was significant.(2)PQ test of digestive tract and kidney tissue showed that PQ+AH2QDS group was significantly Lower than PQ group(Figure1.3),with significant statistical significance.In PQ group,some rats had renal swelling,purplish red and subcutaneous congestion.HE staining showed extensive bleeding and congestion in renal tissue,tube type,some of the glomeruli are enlarged and deformed,vacuolar degeneration of renal tubular epithelial cells and changes of glomerular crescent body in PQ group.The pathological changes of PQ+Sivelestat group and PQ+AH2QDS group were Lighter(P<0.05).(3)On the third day of the poisoning,immunofluorescence of Apelin protein in renal tissues showed that the PQ+AH2QDS group was higher than that in PQ group,with statistical significance.The Levels of IL-6 and TNF-α in PQ group were higher than those in PQ+AH2QDS group,and the protein expressions of NF-κBp65,Caspase 1,Caspase 8,GRP78 and CHOP were higher than those in PQ+AH2QDS group(P<0.05);The expression of Apelin/APJ in PQ+AH2QDS group was higher than that in PQ+Sivelestat and PQ groups(P<0.05),and increased gradually on day 3 and 7.(4)In cell experiments,CCK8 detected PQ decreased cell viability,while Sivelestat and AH2QDS increased cell viability(P<0.05).Meanwhile,the contents of IL-6,TNFα,OH·,ROS in PQ+AH2QDS and PQ+Sivelestat groups were Lower than those in PQ group(P<0.05).Conclusion:Experiments showed that AH2QDS could directly or indirectly reduce oxidative stress,inflammation,apoptosis and other pathological processes caused by PQ poisoning,and alleviating renal tissue damage.The protective effect of AH2QDS was correlated with up-regulation of Apelin/APJ expression.