Background
The morbidity of nephrolithiasis is 2–3 times higher in males than in females, suggesting that androgen plays a key role in nephrolithiasis. The death of renal tubular epithelial cells (TECs) is an important pathophysiological process contributing to the development of nephrolithiasis. Therefore, the aim of this study is to investigate whether androgen directly induces TECs apoptosis and necrosis and its underlying mechanisms in kidney stone formation.
Materials and methods
We compared serum testosterone level between male and female healthy volunteers and kidney stone patients. The in vivo nephrolithiasis model was established using glyoxylic acid, and calcium deposits were detected by van Kossa staining. In the in vitro study using mouse TECs (TCMK-1 cells) and human TECs (HK-2 cells), apoptosis, necrosis, and the expression of BH3-only protein Bcl-2-like 19 kDa-interacting protein 3 (BNIP3) were examined incubated with different doses of testosterone using flow cytometry. Levels of apoptosis-related proteins transfected with the BNIP3 siRNA were examined by western blotting. The mitochondrial potential (ΔΨm) was detected by JC-1 staining and flow cytometry. We monitored BNIP3 expression in the testosterone-induced TECs injury model after treatment with hypoxia inducible factor 1α (HIF-1α) and/or hypoxia inducible factor 2α (HIF-2α) inhibitors to determine the upstream protein regulating BNIP3 expression. Additionally, ChIP and luciferase assays were performed to confirm the interaction between HIF-1α and BNIP3.
Results
Both male and female patients have significantly higher testosterones compared with healthy volunteers. More calcium deposits in the medulla were detected in male mice compared to female and castrated male mice. Testosterone induced TECs apoptosis and necrosis and increased BNIP3 expression in a dose-dependent manner. Testosterone also increased Bax expression, decreased Bcl-2 expression and induced a loss of ΔΨm. This effect was reversed by BNIP3 knockdown. HIF-1α inhibition significantly decreased BNIP3 expression and protected TECs from testosterone-induced apoptosis and necrosis. HIF-2α inhibition, however, did not influence BNIP3 expression or TECs apoptosis or necrosis. Finally, HIF-1α interacted with the BNIP3 promoter region.
Conclusion
Based on these results, testosterone induced renal TECs death by activating the HIF-1α/BNIP3 pathway.
Electronic supplementary material
The online version of this article (10.1186/s12967-019-1821-7) contains supplementary material, which is available to authorized users.
An ideal bone repair scaffold is expected to possess superior architectural characteristics to facilitate the adhesion, proliferation, and migration of bone‐repair‐related cells, while excluding nonosteogenic cells and fibrous tissues from interfering with normal bone regeneration. Unfortunately, such scaffold material has rarely been reported. Herein, nanocomposite scaffolds with a radially ordered porous structure are presented, manufactured using a modified directional freeze‐casting method, and are promising bone defect repair materials to satisfy this requirement. The prepared nanocomposite scaffolds consist of a natural bio‐macromolecule, chitosan, and bioactive hydroxyapatite nanoparticles derived from porcine cortical bone, demonstrating favorable biocompatibility and biological functions. Both in vitro cell studies and in vivo animal studies reveal the great superiority of the radially oriented porous structure of the scaffolds in guiding bone regeneration, while simultaneously preventing the invasion of surrounding nonosteogenic cells and fibrous tissue, compared to the axially oriented porous structure. This work indicates the distinctive potential of radially oriented porous scaffolds for repairing tabular and lacunar bone defects.
Background
Endoplasmic reticulum (ER) stress has been found to foster the escape of cancer cells from immune surveillance and upregulate PD-L1 expression. However, the underlying mechanisms are unknown.
Methods
While analyzing the protein levels using immunofluorescence and Western blotting, the RNA levels were measured using qRT-PCR. Ten injection of exosomes into six-week-old nude mice was made through the tail vein once every other day in total.
Results
The expression of certain ER stress markers such as PERK (PKR-like endoplasmic reticulum kinase), ATF6 (activating transcription factor 6), and GRP78 (glucose-regulated protein 78), was found to be upregulated in the oral squamous cell carcinoma (OSCC) tissues and related to poor overall survival. There is a positive relationship between the extent of ER stress-related proteins and a cluster of PD-L1 expression and macrophage infiltration among the OSCC tissues. Further, incubation with exosomes derived from ER-stressed HN4 cells (Exo-ER) was found to upregulate PD-L1 extents in macrophages in vitro and in vivo, and macrophage polarization toward the M2 subtype was promoted by upregulating PD-L1.
Conclusions
ER stress causes OSCC cells to secrete exosomal PD-L1 and upregulates PD-L1 expression in macrophages to drive M2 macrophage polarization. The delineation of a new exosome-modulated mechanism was made for OSCC–macrophage crosstalk driving tumor development and to be examined for its therapeutic use.
Graphical abstract
Exosomal PD-L1 secreted by ER-stressed OSCC cells promoted M2 macrophage polarization.
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