RESUMEN
Purpose: This study aims to explore the effect and underlying mechanism of Chonggu Granules (CGG) in knee osteoarthritis (KOA) in rats. Methods: A papain-induced KOA model was established in rats. The pathological alterations of extracellular matrix in rat cartilage tissues were observed through hematoxylin and eosin (H&E) staining, followed by Mankin score for quantitative scoring. The ultrastructure of cartilage extracellular matrix was examined under a transmission electron microscopy (TEM). ELISA was used to measure the levels of IL-6, TNF-α, and IL-1ß in rat serum. Immunofluorescence was performed for assessing the levels of MMP-3, MMP-13, and Col2al in rat cartilage. Western blot was used to identify the protein expressions of wnt1, GSK-3ß, ß-catenin, and Aggrecan in rat cartilage. The mRNA relative expressions of miR-148a-3p, wnt1, ß-catenin, and GSK-3ß in rat cartilage were detected by RT-PCR. Luciferase reporter gene was used to detect the target genes of miR-148a-3p. Results: CGG significantly improved articular cartilage tissue and extracellular matrix metabolism compared to the model group as indicated by H&E, Mankin score, and TEM data. Moreover, low, medium, and high doses of CGG reduced the levels of IL-6, TNF-α, IL-1ß, MMP-3, and MMP-13 in serum to varying degrees but increased the levels of Col2al and Aggrecan. Mechanistically, CGG targeted wnt1 by increasing the expression of miR-148a-3p in a dose-dependent manner, thereby downregulating the mRNA and protein expressions of ß-catenin in cartilage tissue and upregulating the mRNA and protein expressions of GSK-3ß. Conclusion: CGG may control the miR-148a-3p/wnt/ß-catenin signaling pathway to decrease the levels of its downstream target genes MMP-13 and MMP-3, increase the expressions of Col2al and Aggrecan, and downregulate the contents of inflammatory cytokines IL-6, TNF-α, and IL-1ß, thereby improving the metabolism of cartilage extracellular matrix and alleviating the degeneration of articular cartilage in KOA.
RESUMEN
Systemic lupus erythematosus (SLE) is a chronic disease of the autoimmune system with multiple damages, most commonly renal damage. The aim of the present study is to examine the therapeutic ability of Qihuang Jianpi Zishen decoction (QJZ) on MRL/lpr mice and uncover its mechanism preliminarily. Twenty-four female MRL/lpr mice were assigned into the model, prednisolone, mycophenolate mofetil and QJZ groups randomly. Six C57BL/6 mice were considered as controls. Each group was treated with corresponding drugs for 4 weeks, anti-dsDNA autoantibodies, C3 and C4, renal function and renal histopathological changes were observed. The expression of GAS5/miR-21/sprouty1 axis and ERK/CREB pathway in kidney was identified by western blotting and qRT-PCR. Compared with MRL/lpr mice, anti-dsDNA autoantibodies of mice treated with QJZ were significantly down-regulated, C3 and C4 were significantly up-regulated. QJZ also alleviated proteinuria, decreased SCr and BUN levels and minimized renal histopathological changes. In addition, QJZ affected the expression of GAS5/miR-21/sprouty1 axis and the phosphorylation of ERK/CREB pathway in renal tissues. QJZ bears therapeutic ability on healing renal injury in MRL/lpr mice. These effects may be achieved by regulating the GAS5/miR-21/sprouty1 axis and inhibiting the ERK/CREB pathway, thus improving the excessive proliferation of glomerular mesangial cells.
