Shock Wave Therapy Enhances Mitochondrial Delivery into Target Cells and Protects against Acute Respiratory Distress Syndrome

Abstract

This study tested the hypothesis that shock wave therapy (SW) enhances mitochondrial uptake into the lung epithelial and parenchymal cells to attenuate lung injury from acute respiratory distress syndrome (ARDS). ARDS was induced in rats through continuous inhalation of 100% oxygen for 48 h, while SW entailed application 0.15 mJ/mm2 for 200 impulses at 6 Hz per left/right lung field. In vitro and ex vivo studies showed that SW enhances mitochondrial uptake into lung epithelial and parenchyma cells (all p < 0.001). Flow cytometry demonstrated that albumin levels and numbers of inflammatory cells (Ly6G+/CD14+/CD68+/CD11b/c+) in bronchoalveolar lavage fluid were the highest in untreated ARDS, were progressively reduced across SW, Mito, and SW + Mito (all p < 0.0001), and were the lowest in sham controls. The same profile was also seen for fibrosis/collagen deposition, levels of biomarkers of oxidative stress (NOX-1/NOX-2/oxidized protein), inflammation (MMP-9/TNF-α/NF-κB/IL-1β/ICAM-1), apoptosis (cleaved caspase 3/PARP), fibrosis (Smad3/TGF-β), mitochondrial damage (cytosolic cytochrome c) (all p < 0.0001), and DNA damage (γ-H2AX+), and numbers of parenchymal inflammatory cells (CD11+/CD14+/CD40L+/F4/80+) (p < 0.0001). These results suggest that SW-assisted Mito therapy effectively protects the lung parenchyma from ARDS-induced injury.

Hepatic31P‐magnetic resonance spectroscopy identified the impact of melatonin‐pretreated mitochondria in acute liver ischaemia‐reperfusion injury

“…In this way, our findings, in addition to being consistent with the findings from previous studies, [45][46][47][48][49] could, once again, explain why the liver enzyme, fibrosis and liver injury score were markedly increased in IRI animals. Of importance was that melatonin-pretreated Mito therapy significantly suppressed inflammatory reaction, DNA damage, oxidative stress, cellular apoptosis and fibrosis.…”

“…Plentiful data have shown that exogenous mitochondrial transfusion not only effectively rescued the function and integrity of endogenous mitochondria but also refreshed the depleted mitochondria in organ, resulting in preserving the ischaemia-reperfusion-related organ dysfunction. 16,46 The most important finding in the present study was that melatonin-pretreated exogenous mitochondria not [45][46][47][48][49] Additionally, increased oxidative stress in circulation and intracellular/mitochondrial compartment notably down-regulated mitochondrial function and depleted energy-storage capacity. 16,46 An essential finding in the present study was that these above-mentioned parameters of molecular-cellular perturbations [45][46][47][48][49] were substantially increased in IRI animals as compared to SC animals.…”

“…The dosage of mitochondria utilized in the present study was based on our previous studies with some modifications. 42,45,46 The procedure was described as follows: the left lobe of the liver was dissected free from the surrounding ligaments. Hepatic ischaemia was induced by temporal obstruction of the vessels by placing a 4-0 silk loop around the hilar region of the left liver lobe in groups 2 and 3 animals, whereas SC animals received only laparotomy without undergoing hepatic ischaemia.…”

“…16,42 The procedure and protocol of liver mitochondrial isolation from donor SD rats have been previously described by our studies. 42,45,46 In detail, the rats (ie the donors) were fasted overnight prior to the mitochondrial isolation procedure, then killed and their gallbladders and livers carefully isolated and removed. Immediately, the liver (3 g) was immersed in 50 mL of ice-cold IBc (10 mmol/L Tris-MOPS, 5 mmol/L EGTA/Tris and 200 mmol/L sucrose, pH 7.4) in a beaker, followed by rinsing the liver free of blood with ice-cold IBc.…”

“…The procedure and protocol for Western blot analysis have been described in detail in our previous studies. 42,45,46 Briefly, equal amounts…”