Differential Gene Expression of PI3K/AKT Signaling Pathway in the Treatment of Ulcerative Colitis by Dangshen(党参)
Objective:To confirm the differential expression of genes in the phosphatidylinositol 3 kinase(PI3K)/protein kinase B(Akt)signaling pathway in the mouse model of ulcerative colitis(UC)treated by Dangshen(党参)based on the transcriptomic data.Method:SD rats were randomized into a blank group and a modeling group.The mouse model of UC was established by the 2,4,6-trinitrobenzenesulfonic acid(TN-BS)-ethanol method.The successfully modeled rats were randomized into model,positive drug(salazosulfapyridine,SASP,0.3 g/kg),and Dangshen(18,9,and 4.5 g/kg)groups.After 7 consecutive days of drug intervention,the serum and colon tissue samples of each group were collected.The disease activity index(DAI)was calculated.The pathological sections were stained with hematoxylin-eosin(HE),and the tis-sue damage index(TDI)of the colon mucosa and the colon index were calculated.Enzyme-linked immunosorbent assay was employed to measure the levels of interleukin-1[3(IL-1β),IL-6,IL-8,tumor necrosis factor-α(TNF-α),calcitonin(PCT),and C-reactive protein(CRP)in the serum.Transcriptional sequencing was carried out to analyze the differentially expressed genes(DEGs)in the colon tissue a-mong the blank,model,and Dangshen(18 g/kg)groups.Gene Ontology(GO)annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were performed to analyze the signaling pathway in DEGs.Western blotting(WB)was employed to determine the protein levels of Akt and PI3K in the colon tissue.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was em-ployed to measure the mRNA levels of Akt,Pi3k,collagen type Ⅳ α-6(Col4a6),laminin α2(Lama2),nerve growth factor receptor(Ngfr),laminin subunit γ3(Lamc3),laminin β2(Lamb2),integrin subunit α7(Itga7),lysophosphatidic acid receptor 3(Lpar3),ephrin A1(Efna1),prolactin receptor(Prlr),3-phosphoinositide-dependent protein kinase 1(Pdpk1),and cyclin d2(Ccnd2)in the colon tissue.Re-sults:Compared with the blank group,the modeling decreased the body weight(P＜0.01),shortened the colon(P＜0.01),and increased the DAI,colon index,and colon TDI(P＜0.01).Compared with the model group,the drug interventions increased the body weight(P＜0.01),extended the colon(P＜0.01 or P＜0.05),and decreased the DAI,colon index,and colon TDI(P＜0.01 or P＜0.05).The 4 727 DEGs be-tween the model group and the blank group,as well as the 1 058 DEGs between the drug intervention group and the model group,were signifi-cantly enriched in the PI3K/Akt signaling pathway.Among them,Col4a6,Lama2,Ngfr,Lamc3,Lamb2,Itga7,Lpar3,Efna1,Prlr,Pdpk1,and Ccnd2 showed significantly differential expression.Compared with the blank group,the modeling elevated the levels of IL-1β,IL-6,IL-8,TNF-α,PCT,and CRP in the serum(P＜0.01),up-regulated the protein levels of PI3K and Akt and the mRNA levels of Pi3k,Akt,Pdpk1,and Ccnd2 in the colon tissue(P＜0.01),and down-regulated the mRNA levels of Col4a6,Lama2,Ngfr,Lamc3,Lamb2,Itga7,Lpar3,Efna1,and Prlr in the colon tissue(P＜0.01).Compared with the model group,the drug intervention lowered the levels of IL-1β,IL-6,IL-8,TNF-α,PCT,and CRP in the serum(P＜0.01),down-regulated the protein levels of PI3K and Akt and the mRNA levels ofPi3k,Akt,Pdpk1,and Ccnd2 in the colon tissue(P＜0.01 or P＜0.05),and up-regulated the mRNA levels of Col4a6,Lama2,Ngfr,Lamc3,Lamb2,Itga7,Lpar3,and Prlr in the colon tis-sue(P＜0.01 or P＜0.05).Conclusion:Dangshen may exert therapeutic effects on UC by inhibiting the PI3K/Akt signaling pathway and regulating the expression of Col4a6,Lama2,Ngfr,Lamc3,Lamb2,Itga7,Lpar3,Efna 1,Prlr,Pdpk 1,and Ccnd2.Using transcriptomic to explore the significant DEGs provides a new research perspective for deciphering the mechanism of Dangshen in treating UC.
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