Restoration of Hepatic and Intestinal Integrity by Phyllanthus amarus Is Dependent on Bax/Caspase 3 Modulation in Intestinal Ischemia-/Reperfusion-Induced Injury

Abstract

Ethnopharmacological relevance: Oxidative stress is a key player in intestinal ischemia/reperfusion (I/R) injury (IIRI) with a tendency to trigger systemic inflammatory response, resulting in progressive distal organ injury. To date, the role of Bax/caspase 3 signaling in IIRI has not been reported. Furthermore, the discovery of a safe and effective drug remains pertinent in improving the outcome of IIRI. Therefore, this study investigated the role of Bax/caspase 3 signaling in intestinal I/R-induced intestinal and hepatic injury. In addition, the protective effect and possible associated mechanism of action of methanolic Phyllanthus amarus leaf extract (PA) against intestinal I/R-induced intestinal and hepatic injury were evaluated. Materials and methods: Fifty male Wistar rats were randomized into five groups (n = 10). The sham-operated group was received 0.5 mL of distilled water for seven days prior to the sham surgery, while the IIRI, febuxostat (FEB) + IIRI, low-dose PA (LDPA) + IIRI, and high-dose PA (HDPA) + IIRI groups underwent the I/R procedure. In addition to the procedure, IIRI, FEB + IIRI, LDPA + IIRI, and HDPA + IIRI received 0.5 mL of distilled water, 10 mg/kg of febuxostat, 200 mg/kg of PA, and 400 mg/kg of PA, respectively, for seven days prior to the I/R procedure. Results: Administration of methanolic Phyllanthus amarus leaf extracts attenuated the intestinal I/R-induced rise in intestinal and hepatic injury markers, malondialdehyde, nitric oxide, TNF-α, IL-6, and myeloperoxidase activities. In addition, Phyllanthus amarus ameliorated I/R-induced suppression of reduced glutathione, thiol and non-thiol proteins, and superoxide dismutase, catalase, and glutathione peroxidase activities in intestinal and hepatic tissues. These were coupled with the suppression of I/R-induced bacterial translocation, downregulation of I/R-induced activation of Bax/caspase 3 signaling, and improvement of I/R-induced distortion of intestinal and hepatic histoarchitecture by Phyllanthus amarus. Conclusion: Methanolic Phyllanthus amarus leaf extract protects against intestinal and hepatic injuries associated with intestinal I/R by suppressing oxidative-stress-mediated activation of Bax/caspase 3 signaling. The beneficial effects of Phyllanthus amarus may be ascribed to its constituent bioactive molecules, especially tannins, anthocyanin, alkaloids, and phenolics.

When less is more: Experimental Bishop–Koop technique for reduction in the use of laboratory animals for intestinal pathophysiological studies

“…All of these studies required the use of one animal per sampling incident, which highlights the importance of techniques that optimise the use of experimental animals. [20][21][22][23][24][25][26][27][28][29] In our selective clamping IIRI-BK model, we obtained five samples from the same injured intestine, considerably reducing the number of rats and allowing kinetic studies. In addition, SI morphological damage and villus recovery were comparable with several previous results published by our group and others.…”

“…Among experimental studies focused on gastrointestinal diseases found in the literature, IIRI-and 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis in rats appears as a current model used by many authors and research groups around the world. [20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37] For this reason, we selected these two relevant pathologies to evaluate the experimental BK procedure in an intestinal injury scenario. IIRI was used for simulating a model of intestinal intussusception, strangulated hernia or intestinal transplantation.…”

“…For example, in the SI, superior mesenteric artery clamping model is currently used to evaluate IIRI, requiring one animal for each sample and study time. [15][16][17][18][19][20][21][22][23][24][25][26][27][28][29] The same situation occurs in the field of experimental colitis, where old and recent scientific articles use one animal in each LI sample collection. [30][31][32][33][34][35][36][37] For the reasons mentioned above, it is a challenge to develop models that enable reducing the number of animals involved in these studies.…”

Moringa oleifera-based feed supplement protects against renal ischaemia/reperfusion injury via downregulation of Bax/caspase 3 signaling

“…Animals were prepared for surgical induction as earlier stated ( 35 , 36 ). The rats were weighed and anesthetized with 10 mg/kg of Xylazine and 50 mg/kg of Ketamine.…”

Efectos del precondicionamiento isquémico abdominal multiorgánico en modelos experimentales de isquemia-reperfusión

“…De los estudios experimentales focalizados en patología de intestino delgado la IIR es un modelo de interés en la actualidad. (91)(92)(93)(94)(95)(96)(97)(98)(99)(100) Materiales y métodos: Se emplearon ratas Wistar hembras (peso 221 ± 13 gr). A las mismas se les proporcionó agua y alimento a demanda previo al acto quirúrgico.…”