FOXA1-Driven pathways exacerbate Radiotherapy-Induced kidney injury in colorectal cancer.

OBJECTIVE: This study aimed to investigate the role of FOXA1 in acute kidney injury (AKI) induced by radiotherapy in colorectal cancer. Although FOXA1 is known to be aberrantly expressed in malignant tumors, its contribution to AKI remains unclear. This study aimed to explore the involvement of FOXA1 in AKI induced by radiotherapy in colorectal cancer and its influence on the regulation of downstream target genes. METHODS: Firstly, a transcriptome analysis was performed on mice to establish a radiation-induced AKI model, and qPCR was used to determine the expression of FOXA1 in renal cell injury models induced by X-ray irradiation. Additionally, FOXA1 was silenced using lentiviral vectors to investigate its effects on the apoptosis of mice with radiation-induced AKI and HK-2 cells. Next, bioinformatics analysis and various experimental validation methods such as ChIP assays, co-immunoprecipitation, and dual-luciferase reporter assays were employed to explore the relationship between FOXA1 and the downstream regulatory factors ITCH promoter and the ubiquitin ligase-degradable TXNIP. Finally, lentiviral overexpression or knockout techniques were used to investigate the impact of the FOXA1/ITCH/TXNIP axis on oxidative stress and the activation of inflammatory body NLRP3. RESULTS: This study revealed that FOXA1 was significantly upregulated in the renal tissues of mice with radiation-induced AKI and in the injured HK-2 cells. Furthermore, in vitro cell experiments and animal experiments demonstrated that FOXA1 suppressed the transcription of the E3 ubiquitin ligase ITCH, thereby promoting apoptosis of renal tubular cells and causing renal tissue damage. Further in vivo animal experiments confirmed that TXNIP, a protein degraded by ITCH ubiquitination, could inhibit oxidative stress and the activation of NLRP3 inflammasome in the AKI mouse model. CONCLUSION: FOXA1 enhances oxidative stress, cell apoptosis, and NLRP3 inflammasome activation by regulating the ITCH/TXNIP axis, thereby exacerbating radiotherapy-induced AKI.

[A new method for establishing a ventricular fibrillation model by TCEI in Tibetan miniature pig].

OBJECTIVE: To explore an economical, convenient, safe and efficient method for establishing a Tibetan miniature pig model of cardiac arrest (CA). METHODS: Cardiac puncture was performed in 12 Tibetan miniature pigs using two acupuncture needles. One needle was inserted into the fourth intercostal near the right side of the sternum about 3 cm in depth at an angle of 30° to 60° between the chest and the needle, and the depth was adjusted until the handle of the needle vibrated with the heartbeat without premature ventricular contraction on the electrocardiogram; the other was inserted into the subcutaneous tissue of the left armpit about 3 cm in depth without damaging important organs. The handles of the two needles were connected with 9V dry batteries to form a circuit and generate direct current stimulation. Ventricular fibrillation was produced in the pigs to induce CA by stimulation of transcutaneous electrical induction (TCEI) for 3 s, and the success rate of modeling was recorded. After an interval of 4 min without intervention, cardiopulmonary resuscitation (CPR) was performed using the standard Utstein style, and the survival of the pigs after recovery was observed. RESULTS: The success rate of ventricular fibrillation modeling was 91.67% (11/12) using this method, and CPR achieved a success rate of 45.45% (5/11) in these models. The subsequent survival of the pigs was 100% (5/5) at 24 h and 80% (4/5) at 72 h. After observation for 72 h, the resuscitated Tibetan miniature pigs were dissected, and no significant damage was found in the vital organs in the thoracic or abdominal cavities. CONCLUSIONS: We successfully established a model of CA using acupuncture needles and dry batteries in Tibetan miniature pigs, and this method is economical, convenient, safe and efficient.