Effects of Total Flavonoids of Astragalus Complanatus on Renal Interstitial Fibrosis in UUO Mice and its Mechanism Based on Transcriptome Sequencing Technology
Objective:To study the effect of total flavonoids of Astragalus complanatus on renal interstitial fibrosis in mice with unilateral ureteral obstruction(UUO)model and to investigate its mechanism based on transcriptome sequencing.Methods:25 mice were randomly divided into sham operation group,model group,High-dose and low-dose groups of total flavonoids of Astraga-lus complanatus,irbesartan group,with 5 mice in each group.Establishing a UUO model by ligating a unilateral ureter.After model-ing,the drug was given by intraperitoneal injection once a day for 7 consecutive days.After paraffin sectioning of kidney tissue,HE staining and Masson staining were performed to observe the pathological changes of kidney tissues.The expression levels of α-smooth muscle actin(α-SMA)and fibronectin(Fn)in kidney tissue were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot.In addition,6 mice were randomly divided into model group 1 and high-dose total flavonoids of As-tragalus complanatus 1.According to the above method,the mice were modeled and Administered,and then the kidney tissue of mice in each group was isolated,total RNA was extracted and sequenced,followed by analyzing the different expression genes between the two groups,Conducting GO and KEGG pathway enrichment analysis,the above results are used to predict effective differentially ex-pressed genes.Finally,genes were verified by RT-qPCR.Results:The pathological damage of kidney tissue in the High-dose and low-dose groups of total flavonoids of Astragalus complanatus was significantly improved compared with the model group.The mRNA and protein expression levels of α-SMA and Fn in kidney tissue of model group were significantly higher than those of sham group sham operation group(P＜0.01).After treatment with total flavonoids of Astragalus complanatus,the levels of these indexes were significantly decreased compared with the model group(P＜0.01).Transcriptome sequencing results:GO enrichment analysis showed that differentially expressed genes are mainly concentrated in the aspects of cell process,biological regulation and biological process regulation.KEGG pathway enrichment analysis showed differentially expressed genes involve Rap1,Ras,PI3K/Akt,MAPK signaling pathways,etc.Two effective genes were screened by transcriptome sequencing.The results of in vivo verification test:inter-leukin-6(IL-6)was highly expressed in the model group,and was significantly down-regulated after intervention of total fla-vonoids of Astragalus complanatus.vasohibin 1(Vash1)was low expressed in model group and significantly up-regulated after inter-vention with total flavonoids from Astragalus complanatus.The results of in vivo verification were consistent with the sequencing re-sults.Conclusion:Total flavonoids of Astragalus complanatus can improve renal interstitial fibrosis in UUO model mice.and its mech-anism may be related to down-regulating IL-6 expression,up-regulating Vash1 expression,and regulating signal pathways such as Rap1,Ras,PI3K-Akt,MAPK signaling pathways,etc.
Total flavonoids of astragalus complanatusRenal interstitial fibrosisUnilateral ureteral obstructionTran-scriptome sequencing
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