SARS-CoV-2 receptor networks in diabetic kidney disease, BK-Virus nephropathy and COVID-19 associated acute kidney injury

COVID-19 morbidity and mortality is increased in patients with diabetes and kidney disease via unknown mechanisms. SARS-CoV-2 uses angiotensin-converting enzyme 2 (ACE2) for entry into host cells. Since ACE2 is a susceptibility factor for infection, we investigated how diabetic kidney disease (DKD) and medications alter ACE2 receptor expression in kidneys. Single cell RNA profiling of healthy living donor (LD) and DKD kidney biopsies revealed ACE2 expression primarily in proximal tubular epithelial cells (PTEC). This cell specific localization was confirmed by in situ hybridization. ACE2 expression levels were unaltered by exposures to renin angiotensin aldosterone system inhibitors in DKD. Bayesian integrative analysis of a large compendium of public -omics datasets identified molecular network modules induced in ACE2-expressing PTEC in DKD (searchable at hb.flatironinstitute.org/covid-kidney) that were linked to viral entry, immune activation, endomembrane reorganization, and RNA processing. The DKD ACE2-positive PTEC module overlapped with expression patterns seen in SARS-CoV-2 infected cells. Similar cellular programs were seen in ACE2-positive PTEC obtained from urine samples of 13 COVID-19 patients who were hospitalized, suggesting a consistent ACE2-coregulated PTEC expression program that may interact with the SARS-CoV-2 infection processes. Thus SARS-CoV-2 receptor networks can seed further research into risk stratification and therapeutic strategies for COVID-19 related kidney damage. Translational statementTo understand the overwhelming burden of kidney disease in COVID-19, we mapped the expression of the SARS-CoV-2 receptor, ACE2, in healthy kidney, early diabetic (DKD) and COVID-19 associated kidney diseases. Single cell RNA sequencing of 111035 cells identified ACE2 predominantly in proximal tubular epithelial cells. ACE2 upregulation was observed in DKD, but was not associated with RAAS inhibition, arguing against an increased risk of COVID-19 among patients taking RAAS inhibitors. Molecular network analysis linked ACE2 expression to innate immune response and viral entry machinery, thereby revealing potential therapeutic strategies against COVID-19.

[Effects of metabolites of mangrove fungus Xylaria sp. from South China Sea Coast on the activity of acetylcholinesterase in vitro].

OBJECTIVE: To investigate the effects of metabolites of mangrove fungus Xylaria sp. from South China Sea Coast, xyloketal A-D, on the activity of acetylcholinesterase (AChE) in vitro. METHODS: Activity of AChE was determined by a modified method of Ellmen. The selectivity of the compounds for AChE and butyrylcholinesterase (BuChE) was also tested and compared with that of a positive control, velnacrine. The inhibitory properties of xylokets on AChE were characterized as well. RESULTS: AChE activity was inhibited by xyloketal A-D in a dose-dependent manner, their IC50 were determined to be 29.9, 137.4, 109.3 and 425.6 micromol/L, respectively. At the same time, velnacrine and all the four compounds showed inhibitory effects on BuChE in different degree, and the inhibitory activity of xyloketals on AChE was found to be reversible and noncompetitive. CONCLUSION: Xyloketal A-D can inhibit AChE as well as BuChE activity in vitro. So xyloketal A-D were likely considered as drug candidates against Alzheimer' s disease (AD) for further development.

The inhibitive effects of resveratrol on adhesion of neutrophils to HUVECs induced by fMLP / 中国药理学通报

Aim To study the effects of resveratrol(Res)on the release of soluble adhesion molecules from human umbilical vein endothelial cells(HUVECs)induced by N-formyl-methionyl-leucyl-phenylal-anine(fMLP)and the adhesion between neutrophils and HUVECs.Methods The effects of Res on neutrophils adhesion to human umbilical endothelial cell(HUVECs)triggered by fMLP were examined.The soluble intercellular cell adhesive molecule-1(ICAM-1),the soluble vascular cell adhesive molecule-1(VCAM-1)and E-selectin release from fMLP(10 ?mol?L-1)stimulated HUVECs were determined by ELISA kits.The neutrophils isolated from human vein blood were loaded with Fluo-3,a fluorescent indicator,to detect intracellular free calcium concentration([Ca2+]i),and CLA was used as a chemiluminescent indicator to determine superoxide production in neutrophils.Results Res(1~50 ?mol?L-1)significantly inhibited neutrophil adhesion to fMLP-stimulated HUVECs and also obviously downregulated the levels of ICAM-1,VCAM-1 and E-selectin in supernatant of HUVECs culture stimulated by fMLP in the dose-dependent pattern.Res also suppressed fMLP-activated superoxide generation and[Ca2+]i increase in neutrophils.Conclusions Res involved in neutrophil adhesion to HUVECs intermediated by cell adhesive molecules expression trigged by[Ca2+]i and superoxide production in neutrophils,which means a lot to prevent inflammatory diseases.

Effects of cilostazol on LPS-stimulated adhesion and soluble adherent molecules release / 中国药理学通报

AIM To examine the effect of cilostazol, a no vel selective phosphodiesterase type 3 inhibitor, on adherence between neutrophils and human umbilical e ndothelial cells ( HUVECs ) and investigate its possible mechanisms. MET HODS Confluent HUVECs between 4～6 passages were used and stimulated by l ipopolysaccharide (LPS, 5 mg?L -1 ) with or without coincubation of cilosta zol (1～10 ?mol?L -1 ) for 24 h. Soluble cell adhesion molecules (sCAMs), including vascular cell adhesion molecule-1 (sVCAM-1), intercellular adhesion molecule-1 (sICAM-1) and endothelial leukocyte adhesion molecule-1 (sELAM-1, sE-selectin) in cell culture medium were measured by ELISA. RESULTS Cilostazol (1～10 ?mol?L -1 ) inhibited adherence between neutrophils and HUVECs in a dose- dependent manor. At the same time, cilostazol didn't affect sICAM-1 and sE-sel ectin release from LPS-stimulated HUVECs, but in contrast, it significantly inh ibited sVCAM-1 production under the same experiment condition, and this effect was canceled by H-89, an inhibitor of protein kinase A ( PKA ). CONCLUS ION Cilostazol significantly inhibits adherence between neutrophils and H UVECs, and downregulates sVCAM-1 release from LPS-activated HUVECs, and these effects on cytokine-challenged endothelial cells might be via a PKA-dependent pathway. The present result suggests that cilostazol partially eliminates some o f the adherent reactions of HUVECs to LPS, a deleterious cytokine, and it is rea sonable to consider that cilostazol might be a strategy for preventing atheroscl erosis and other cardiovascular diseases.