山东医药2024，Vol.64Issue(2)：22-26,5.DOI:10.3969/j.issn.1002-266X.2024.02.005
龙胆苦苷腹腔注射对单侧输尿管梗阻小鼠肾间质纤维化的改善作用及其机制
Effect and mechanism of intraperitoneal injection of gentiopicroside on renal interstitial fibrosis in UUO mice
摘要
Abstract
Objective To investigate the effect and mechanism of gentiopicroside(GPS)on renal interstitial fibrosis in unilateral ureteral obstruction(UUO)mice.Methods Twenty-five C57BL/6 male mice were randomly divided into the sham operation group,model control group,low-dose GPS group,high-dose GPS group,and irbesartan group,with 5 mice in each group.Mice in the sham operation group were fed normally,while in the other four groups,we established the UUO models by ligating one side of the ureter in mouse.After successful modeling,mice in the low-dose and high-dose GPS groups were given 0.1 and 0.2 g/kg GPS through intraperitoneal injection,mice in the irbesartan group were given 0.02 g/kg irbesartan intraperitoneally,and mice in the model group and sham operation group were given equal volume of normal saline through intraperitoneal injection for 7 consecutive days.The mice were executed to take the kidney tissues to observe the morphological changes of kidney tissues using HE staining,and to observe the fibrosis of kidney tissues using Masson staining.Western blotting and RT-qPCR were used to detect the protein and mRNA expression levels of fibronectin(Fn)and α-smooth muscle actin(α-SMA)in the renal tissues.Another six mice were randomly divided into the model group and intervention group,with 3 mice in each.After the UUO models were established,the intervention group was giv-en 0.2 g/kg GPS through intraperitoneal injection,while the model group was given equal volume of normal saline intraper-itoneally for 7 consecutive days.Mice were euthanized,the kidney tissues were taken for transcriptome sequencing,two sets of differentially expressed genes were analyzed,and GO and KEGG pathways enrichment were conducted.Finally,RT-qPCR was used for validation.Results Compared with the sham operation group,the model control group showed significant dilation of renal tubules,a large number of inflammatory cells in the renal tissues,and deposition of a large amount of collagen fibers in the renal interstitium.Compared with the model control group,the low-dose and high-dose GPS groups showed a certain degree of reduction in the renal tubular dilation and inflammatory cell infiltration in the kid-ney tissues,and improved renal interstitial fibrosis,with more significant effect in the high-dose GPS group.Compared with the sham operation group,the Fn,α-SMA protein and mRNA expression levels increased in the renal tissues of the model control group.Compared with the model control group,the Fn,α-SMA protein and mRNA expression levels de-creased in the renal tissues of the low-dose and high-dose GPS groups(all P<0.05),and those in the high-dose group were lower than those of the low-dose group.The sequencing analysis results showed that there were 710 differentially expressed genes between the model group and the intervention group.GO enrichment analysis showed that differentially expressed genes were mainly concentrated in cell processes,biological regulation,and biological process regulation;KEGG pathway enrichment analysis revealed that differentially expressed genes involved Ras,Rap1,PI3K/AKT,MAPK signaling path-ways,etc.Two effective genes HGF and CCDC3 were screened for validation.Compared with the model group,the expres-sion of HGF and CCDC3 in the intervention group increased(both P<0.05),which was consistent with the sequencing re-sults.Conclusion GPS can effectively alleviate renal fibrosis in UUO mice,and its mechanism of action may be related to up-regulating the expression of HGF and CCDC3,Rap1,Ras,PI3K/AKT,MAPK signaling pathways,and other bio-logical pathways.关键词
龙胆苦苷/肾间质纤维化/纤连蛋白/α平滑肌肌动蛋白/输尿管梗阻/转录组测序Key words
gentiopicroside/renal interstitial fibrosis/fibronectin/α-smooth muscle actin/ureteral obstruction/transcriptome sequencing分类
医药卫生引用本文复制引用
彭炳鸿,曾琪,谭睿陟,粟宏伟,刘建..龙胆苦苷腹腔注射对单侧输尿管梗阻小鼠肾间质纤维化的改善作用及其机制[J].山东医药,2024,64(2):22-26,5.基金项目
四川省科技计划项目(2022YFH0118,2022YFS0621). （2022YFH0118,2022YFS0621）
