RESUMO
OBJECTIVE Chronic kidney disease (CKD) has become a global public health problem with 10％-15％incidence rate, and inhibiting the renal interstitial fibrosis is considered to be a potential strategy to delay the progression of CKD. Z-Guggulsterone (Z-GS), an active compound from derived from Commiphora mukul, has been proved to be effective in various diseases. The present study aimes to determine the protective effect and the molecular mechanism of Z-GS on renal fibrosis. METHODS Unilateral ureteral obstruction (UUO) mice and hypoxia-induced HK-2 cells were used to simulate renal fibrosis in vitro and in vivo, respectively. The mice and cells were treated with different doses of Z-GS to observe the pharmacological action. Renal function, including Scr, BUN, and UA, were detected by commercial kits. H&E and Masson staining were performed to observe histopathological changes of kidney. Cell viability and LDH release of HK-2 cells were detected by commercial kits. Cell cycle distribution and apoptosis rate were analyzed by flow cytometry. Fibrosis markers were detected by immunohistochemistry and immunofluorescence analysis. Cell cycle related proteins and Klotho/p53 signaling were analyzed by Western blotting. RESULTS The results showed that Z-GS decreased the rise of Scr, BUN, and UA and lightened renal histopathological injury, which were induced by UUO. Besides, Z-GS administration alleviated renal fibrosis in mice by inhibiting the expressions of α-SMA, TGF-β and colla?genⅣ, and delayed G2/M cell cycle arrest by promoting the expressions of CDK1 and cyclinD1/B1 rate. Experiments in vitro indicated that Z-GS treatment significantly increased the cell viability while decreased the LDH release in hypoxia-induced HK-2 cells. In addition, hypoxia induced fibrosis and G2/M cycle arrest in HK-2 cells were retarded by Z-GS. The study of its possible mechanism exhibited that Z-GS treatment increased the level of Klotho and inhibited P53 level. Nev?ertheless, the effect of Z-GS on Klotho/P53 signaling was reversed by siRNA-Klotho. Moreover, siRNA-Klotho treatment eliminated the effects of Z-GS on G2/M cell cycle arrest and fibrosis. CONCLUSION This study clarified that Z-GS allevi?ated renal fibrosis and G2/M cycle arrest through Klotho/P53 signaling pathway. People who have suffered CKD may potentially benefit from treatment with Z-GS.
RESUMO
To identify and verify the active ingredients from Astragalus membranaceus on hypertensive cardiac remodeling based on network pharmacology and heart RNA-sequencing data. The monomers of A. membranaceus and their intervention target database were established by using network pharmacology. The genes associated to cardiac remodeling were then screened by analyzing cardiac RNA-sequencing data. An overlap between genes related to cardiac remodeling and targets of ingredients form A. membranaceus was collected to obtain monomers with protective effect on hypertensive cardiac remodeling. Angiotensin Ⅱ(AngⅡ)-induced mouse cardiac remodeling model was used to validate the protective effect of active ingredients from A. membranaceus on hypertensive cardiac remodeling. Finally, a total of 81 monomers and 1 197 targets were enrolled in our database. Mouse RNA-sequencing data showed that 983 genes were significantly up-regulated and 465 genes were down-regulation in myocardial tissues of the cardiac remodeling mice as compared with blank group mice, respectively. Ninety-two genes were found via overlapping between genes related to cardiac remodeling and targets, involving 59 monomers from A. membranaceus. Further research found that vanillic acid(VA) could intervene 27 genes associated with hypertensive cardiac remodeling, ranking top 1. Meanwhile, VA could significantly inhibit AngⅡ-induced increase in ratio of heart weight to body weight and heart weight to tibial length, ANP and BNP mRNA levels in myocardial tissues, myocardial tissue damage, cardiac fibrosis level and cardiac hypertrophy level in vivo. Those results showed that network pharmacology screen-based VA has protective effect on AngⅡ-induced cardiac remodeling.