#3651 Extracellular vesicles from the <i>in vitro</i> AKI model promote cardiac cells stress
Bibliographic record
Abstract
Abstract Background and Aims Extracellular vesicles (EVs) are blebs released by all cell types, exerting intercellular communication function due to their cargo. EVs have been reported to play a role in the pathophysiology of several diseases, such as cancer, cardiovascular, and renal diseases. However, whether EVs contribute to the pathological crosstalk between the kidney and the heart is unknown. We aimed to characterize EVs from in vivo and in vitro acute kidney disease models and to assess whether they would impact cardiac cellular viability. Method C57Bl/6 male mice (uOttawa Animal Facility Protocol: OHRIe3961-A1 and UFABC animal facility commenting number: 9019180122) left kidney were submitted to 60 minutes of warm ischemia and 1 and 8 days of reperfusion (IR1 and IR8). The sham animals (control) only had the left flank opened. HK-2 cells were exposed to hypoxia for 24 hours, followed by 3 hours of reperfusion (HR). The normoxia cells were kept under 21% O2 throughout the HR period. The AKI in vivo model was validated by histological analysis stained with PAS, BUN, and qPCR for KIM-1, and the HR in vitro models were validated by western blot to HIF-1α. EVs were isolated by ultracentrifugation from the plasma and conditioned medium, and characterized by NTA. H9C2 cells were treated with 6000 EVs/ cell from both biofluids for 48 hours, followed by MTT assays. The results were expressed as mean ± standard deviation and analyzed by t-test and ANOVA, in which p-values < 0.005 were considered statistically significant. Results The IR results indicated that in 24 hours of reperfusion, there was a high acute tubular necrosis in the left kidney (0% vs 60%) followed by tubular atrophy and interstitial fibrosis in 8 days (0% vs 73.66%), KIM-1 was upregulated in the left kidney of both 1 and 8 days of reperfusion groups (Sham: 1 ± 1.22 vs IR1: 434.80 ± 192.55, and IR8d: 162.22 ± 84.74), and there was an impairment of the kidney function in 8 days of reperfusion (Sham: 1.66 ± 0.22 vs IR8d: 2.15 ± 0.15). HK-2 submitted to HR expressed 2-fold more HIF-1α than normoxia cells (Normoxia: 1.00 ± 0.11 vs Hypoxia: 1.93 ± 0.81). Regarding EVs characterization, the IR protocol did not induce changes in the total particles/mL (IR1: 2.54 ± 0.52 and IR8: 2.03 ± 0.51 vs Sham: 2.43 ± 0.54) but promoted the particle size (nm) decrease (IR1: 126.33 ± 2.37 and IR8: 128.90 ± 0.86 vs Sham:154.90 ± 4.80). HR-induced HK-2 cells release 2-fold more EVs with bigger sizes (nm) than the control group (Hypoxia: 14.52 ± 7.47 vs Normoxia: 5.30 ± 1.34 particles/mL) (Hypoxia: 183.92 ± 3.90 nm vs Normoxia: 159.55 ± 10.20 nm). Plasma EVs did not affect the cellular viability of H9C2 cells (Sham: 0.94 ± 0.07, IR1: 1.04 ± 0.10 and IR8: 0.98 ± 0.98 vs Untreated: 1.00 ± 0.09), nevertheless, both normoxic and hypoxic EVs induced the reduction of the cellular viability (Normoxia: 0.85 ± 0.02 and Hypoxia: 0.90 ± 0.04 vs Untreated: 1.00 ± 0.04). Conclusion The present results indicated the accomplishment of the IR model as well as HR. The characterization results suggested that hypoxia stimulates EVs biogenesis, release in renal epithelial cells, and also modification in the size. The MTT results indicated that EVs caused stress in cardiac cells affecting their viability.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.001 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.001 | 0.001 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.004 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".