Tangshen formula attenuates diabetic renal injuries by upregulating autophagy via inhibition of PLZF expression

Abstract

The Chinese herbal granule Tangshen Formula (TSF) has been proven to decrease proteinuria and improve estimated glomerular filtration rate (eGFR) in diabetic kidney disease (DKD) patients. However, the underlying mechanism of TSF on treatment of diabetic nephropathy (DN) remains unclear. The present study aimed to identify the therapeutic target of TSF in diabetic renal injuries through microarray-based gene expression profiling and establish its underlying mechanism. TSF treatment significantly attenuated diabetic renal injuries by inhibiting urinary excretion of albumin and renal histological injuries in diabetic (db/db) mice. We found that PLZF might be the molecular target of TSF in DN. In vivo, the db/db mice showed a significant increase in renal protein expression of PLZF and collagen III, and decrease in renal autophagy levels (downregulated LC3 II and upregulated p62/SQSTM1) compared to db/m mice. The application of TSF resulted in the downregulation of PLZF and collagen III and upregulation of autophagy level in the kidneys of db/db mice. In vitro, TSF reduced high glucose (HG)-induced cell proliferation for NRK52E cells. Further studies indicated that the exposure of NRK52E cells to high levels of glucose resulted in the downregulation of cellular autophagy and upregulation of collagen III protein, which was reversed by TSF treatment by decreasing PLZF expression. In conclusion, TSF might have induced cellular autophagy by inhibiting PLZF expression, which in turn resulted in an increase in autophagic degradation of collagen III that attenuated diabetic renal injuries.

Tangshen Formula Attenuates Diabetic Nephropathy by Promoting ABCA1-Mediated Renal Cholesterol Efflux in db/db Mice

“…The cells were divided into the following groups: the OA group and the PA group, which were grown with 50 μM oleic acid and 50 μM sodium palmitate, respectively. The PA+TSF 250 and PA+TSF 500 groups were treated with 50 μM sodium palmitate + 250 μg/mL TSF and 50 μM sodium palmitate + 500 μg/mL TSF, respectively (Zhao et al, 2017 ).…”

“…The degree of glomerulosclerosis was calculated by the percentage of extracellular matrix (ECM) deposition and mesangial expansion and evaluated at 400X power in 20 cortical fields. The mesangial matrix was scored as follows: 1, <10%; 2, 10–25%; 3, 26–50%; 4, 51–75%; 5, 76–95%; 6, >95% (Zhao et al, 2017 ). The renal sections (8 μm) of fixed frozen mouse kidneys were prepared for Oil Red O and filipin cholesterol staining.…”

Tangshen Formula Treatment for Diabetic Kidney Disease by Inhibiting Racgap1-stata5-Mediated Cell Proliferation and Restoring miR-669j-Arntl-Related Circadian Rhythm

“…A recent prospective, multicenter, double-blind, randomized, controlled study has demonstrated the significant benefit of TSF in decreasing proteinuria and improving estimated glomerular filtration rate in DKD patients compared with placebo [ 12 ]. Further studies indicate TSF may attenuate DKD by inhibiting zinc finger and BTB domain containing 16 (ZBTB16/PLZF) expression [ 13 ], transforming growth factor beta (TGF-β)/SMAD family member 3 (Smad3)/nuclear factor kappa B subunit (NF-κB) signaling pathways [ 14 ], and activating Janus kinase (JAK)/signal transducer and activator of transcription (STAT)/suppressor of cytokine signaling (SOCS) signaling pathway [ 15 ]. However, the mechanism of TSF in DKD treatment remains unclear.…”

“…The goal of this study was to further explore the molecular mechanisms of TSF treatment for DKD by collecting microarray dataset GSE90842 from the National Center of Biotechnology Information Gene Expression Omnibus (NCBI GEO) public database [ 13 ] and analyzing it by a series of bioinformatics software. Compared with the study of Zhao et al [ 13 ], a non-strict threshold (P<0.05 and |logFC (fold change) | >0.1 vs. |logFC| >0.5) was selected to obtain more differentially-expressed genes (DEGs), and DEGs were run between the 2 groups by t test but not among the 3 groups by one-way ANOVA. The crucial DEGs were subsequently identified by construction of a protein and protein interaction (PPI) network and module analysis.…”

“…The underlying microRNAs (miRNAs) regulating DEGs and the small molecular drugs having similar functions to TSF were also predicted. These studies were not performed by Zhao et al [ 13 ]. Our findings may provide new insights into the mechanisms of TSF treatment for DKD.…”