Protective Effect Of Matrine On Acute Kidney Injury Induced By Cisplatin In Mice And Mechanism

Objective: To establish an acute kidney injury mouse model induced by cisplatin and to study the protective effect of matrine on acute kidney injury mouse model induced by cisplatin.The relationship between apoptosis and autophagy and the mechanism of matrine improving acute kidney injury induced by cisplatin in mice was further explored by detecting the related indexes of apoptosis and autophagy.Bioinformatics is used to predict the pathway,which provides a theoretical basis for the next experiment.Methods: 1.Observe the effect of different concentration of cisplatin on kidney.Thirty male C57BL/6 mice were randomly divided into the following 5groups:(1)normal group;(2)Cisplatin 10 mg/kg group;(3)Cisplatin 20 mg/kg group;(4)Cisplatin 30 mg/kg group;(5)Cisplatin 40 mg/kg group.Hematoxylineosin(HE)staining was performed to observe the pathological changes of kidney,observe the effects of different concentrations of cisplatin on the kidney of mice,and explore the optimal injection concentration of cisplatin needed in this study.2.To observe the effect of matrine on acute kidney injury induced by cisplatin in mice.Twenty-four male C57BL/6 mice were randomly divided into the following3 groups :(1)normal control group;(2)Cisplatin group;(3)Matrine + cisplatin group.The mice were sacrificed at day 3 after modeling.The renal function was evaluated by serum creatinine and urea nitrogen detection.Pathomorphological changes of renal tissues were detected by HE staining.Real-time Reverse transcription-Polymerase chain reaction(RT-PCR)technique was used to detect the changes of apoptosis and autophagy in mice kidneys,and immunohistochemical technique was used to detect the related protein expression for apoptosis,autophagy in mice kidneys.Western blotting(WB)technique was used to detect apoptosis and autophagy-related protein expression in mouse kidneys.3.Bioinformatics analysis of matrine in the treatment of acute kidney injury.The target information of acute kidney injury was downloaded from Genecards database to predict the potential target of matrine using Swiss Target Prediction.The target gene information of matrine for the treatment of acute kidney injury can be obtained by intersectional the potential target information of matrine and the target information of acute kidney injury by using the Draw Venn Diagram.The Gene information of the target of matrine for matrine treatment was input into the Profiler for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes,respectively.KEGG pathway enrichment analysis.Results: 1.Serum creatinine and urea nitrogen,HE staining results of renal tissue showed that compared with the normal control group,with the increase of cisplatin injection concentration,the damage of renal tubules in mice was aggravated,the epithelial cells of renal tubules were swollen and necrotic,the tubular type was significantly increased,and the renal function injury was aggravated,and the concentration of 30 mg/kg was the most significant(P< 0.05).2.Compared with cisplatin group,the serum creatinine and urea nitrogen in matrine + cisplatin group were significantly decreased(P< 0.05);He staining of kidney tissue showed that compared with cisplatin group,tubule cells injury in matrine + cisplatin group was significantly reduced(P< 0.05).The results of TUNEL immunofluorescence assay showed that,compared with the cisplatin group,the number of renal apoptotic positive cells decreased in matrine + cisplatin group.RT-PCR results showed that compared with cisplatin group,B cell lymphoma protein-2(Bcl-2)associated X,Bcl-2 associated X,Bcl-2 associated X,MRNA expression of Bax and Cysteine aspartic acid proteinase 3(Caspase3)were significantly decreased.The expression of anti-apoptotic index Bcl-2 and autophagy index Microtubule associated protein 1 light chain 3(LC3)m RNA were significantly increased.Immunohistochemical results showed that,compared with the cisplatin group,the expressions of pro-apoptotic index Bax protein and Caspase3 protein in matrine + cisplatin group were decreased,the protein expressions of anti-apoptotic index Bcl-2 were increased,and the protein expressions of autophagy index LC3 were increased.WB results showed that,compared with cisplatin group,the expression of Bax protein was decreased(P<0.05),the expression of anti-apoptotic index Bcl-2 was increased(P< 0.05),and the autophagy index LC3 was increased(P< 0.05)in matrine + cisplatin group.3.98 corresponding potential matrine targets were identified from Swisstar Get Prediction.Filter Settings(P< 0.05),a total of 48 values were screened out according to the P value.A total of 7040 targets related to acute kidney injury were identified from Gene Cards data.After the intersection of Draw Venn Diagram,a total of 35 target genes were obtained.Thirty-five target genes were imported into the Profiler and analyzed.It was found that the biological process and molecular function of matrine in the treatment of acute kidney injury mainly involved calcium ions or neurotransmitter mediated signal transduction pathways.The main pathways of matrine in the treatment of acute kidney injury include neuroactive ligand-receptor interaction,cyclic adenosine phosphate signaling pathway,calcium signaling pathway,and helper T cell(Th)1/Th2 cell differentiation.Conclusion: 1.In the mouse model of acute kidney injury induced by cisplatin,matrine pretreatment has obvious protective effect on kidney by inhibiting apoptosis and enhancing autophagy inrenal tubular epithelial cells.2.Matrine can protect nephrotoxic kidneys by regulating autophagy and apoptosis by influencing cyclic adenosine phosphate and calcium ion pathways.