Screening And Functional Analysis Of Differentially Expressed Genes In Chronic Glomerulonephritis And Intervention Effect Of Qiteng Xiao Zhuo Granule By Whole Genome Microarray

Aim: Combined with the pharmacodynamics research of Qiteng Xiao Zhuo Granule,to characterize differentially expressed genes in the normal, adriamycin-induced chronic glomerulonephritis(CGN) rats and Qiteng Xiao Zhuo Granule-mediated treatment by genome-wide expression profiles, to evaluate and analyze the molecular mechanisms of CGN and possible roles of Qi Teng Xiao Zhuo granule treatment of CGN.Methods: Adriamycin was injected intravenously into the tail of each rat in the model,Qi Teng Xiao Zhuo granule and Huangkui capsule groups 2 times(n=10); 3.5 mg·kg-1adriamycin was to be given on the first day and 3.0 mg·kg-1 on the fourteenth day. On the twenty-first day, all the rats were put into metabolism cages and urine was collected for 24 hours in order to check 24-hour urinary protein quantitation. A successful model was identified as 24-hour urinary protein quantitation more than 50mg/kg. After model establishment, rats in the Qi Teng Xiao Zhuo granule and Huangkui capsule group were given a gastric gavage of Qi Teng Xiao Zhuo granules(10.8 g·kg-1) and Huangkui capsule(1.0g·kg-1), once per day for 30 days. The normal and model groups were given an equivalent solvent. After the final dose, all the rats were put into metabolism cages and urine was collected for 24 hours. To calculated the kidney visceral index; detected the 24-hour urinary protein, blood urea nitrogen(BUN),serum creatinine(SCr), cystatin c(Cys-c), retinol-binding protein(RBP),?2-microglobulin(?2-MG) by automatic biochemical analyzer; Histological analysis of kidney tissue was by hematoxylin-eosin staining(HE); Total RNA was extrated and subjected to Agilent Rat 4 × 44 K whole genome microarray. Accroding to fold change?2.0, P-value?0.5, Kyoto Encyclopedia of Genes and Genomes(KEGG), Gene Ontology(GO) analyse, LIMMA, String and Cytoscape software were applied to screen and analyze differentially regulated genes. The top 30 co-expression genes of degree among the normal, model, and Qi Teng Xiao Zhuo granule groups were neededto focus on, the real-time PCR(RT-PCR) was performed to verify the co-expression genes.Results: The result of pharmacodynamics was showed that Qi Teng Xiao Zhuo granule could reduced the content of BUN?Cr?Cys-c?RBP??2-MG.The result of gene microarray was showed that 2334 differentially regulated genes were identified including 1294 up-regulated genes and 1040 down-regulated genes between normal and model groups; 563 differentially regulated genes were identified including 342 up-regulated genes and 221 down-regulated genes between model and Qi Teng Xiao Zhuo granule groups; 248 differentially regulated genes were identified including 106 up-regulated genes and 142 down-regulated genes among normal, model and Qi Teng Xiao Zhuo granule groups, which primarily contribute to biological processes such as, cell cycle, inflammatory response, inmmune response, various metabolic process, and so on; pathways such as CytP450 metabolic singaling pathway,B cell receptor singaling pathway, PI3K-Akt singaling pathway, NF-?B singaling pathway, and so on. The top 30 co-expression genes of degree among the normal,model, and Qi Teng Xiao Zhuo granule groups were Fos, Egr1, Syk, Ugt2b15, Hsd3b6,ect, and detection result of RT-PCR was consistent with the result of gene microarray.Conclusion: Whole genome gene expression profile analysis showed that the molecular mechanism of CGN pathogenesis may be related to the promotion of cell cycle, participate in inflammatory response, inmmune response, inhibited anti-inflammatory cytokine and abnormal metabolism; molecular mechanism of Qi Teng Xiao Zhuo granule treatment of CGN may be related to regulated the abnormal genes, inhibited inflammatory response, enhanced the activity of anti-inflammatory cytokine and regulated abnormal metabolism.