Effect of hydrogen-rich water on the chondriosome damage and cytokines in brain tissue of rats with traumatic brain injury

Abstract

To observe the effect of hydrogen-rich water on the chondriosome damage and cytokines change in brain tissue of rats with traumatic brain injury (TBI). Fifty-four health male Sprague-Dawley (SD) rats were divided into three groups by random number table: sham group, trauma group (TBI group), and trauma+hydrogen-rich water group (TBI+HW group), the rats in each group were subdivided into 1, 3 and 7 days subgroups according to the time points after trauma, with 6 rats in each subgroup. The TBI model was reproduced by using a modified Feency method for free fall impact, and the rats in sham group were not given brain impact after craniotomy. The rats in TBI+HW group were given intraperitoneal injection of hydrogen-rich water (5 mL/kg) after TBI model reproduction, and then once a day until being sacrificed; and the rats in sham group and TBI group were given the same amount of normal saline. The neurological severity scores (NSS) for neurologic deficits were calculated at corresponding time points, and then the rats were sacrificed to harvest brain tissue at 3 mm around lesion boundary. The cytokines including tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were determined by enzyme linked immunosorbent assay (ELISA); the protein expressions of Bax, Bcl-2 were determined by Western Blot; the RFU of reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and mitochondrial membrane permeability (MPTP) were determined by fluorescence and enzyme sign method. TBI and TBI+HW groups appeared obvious neurologic damage after injury in rats. NSS scores in TBI and TBI+HW groups showed a decreased tendency with time prolongation after TBI. NSS scores in TBI+HW group at 3 days and 7 days were significantly lower than those of TBI group (NSS score: 9.67±0.82 vs. 11.17±1.17, 6.83±0.75 vs. 8.50±1.04, both P < 0.05). Compared with sham group, the expressions of TNF-α, IL-1β, RFU of ROS in chondriosome, protein expression of Bax in brain tissue in TBI group and TBI+HW group were significantly increased, peaked at 1 day, then they gradually declined. Each time point of RFU of MMP, MPTP in chondriosome and protein expression of Bcl-2 were significantly decreased, and gradually increased after one-day valley value. Compared with TBI group, the expressions of TNF-α, IL-1β, RFU of ROS in chondriosome and protein expression of Bax in brain tissue were all declined at corresponding time points [TNF-α (ng/L): 54.14±1.11 vs. 81.49±2.76, IL-1β (ng/L): 74.53±1.75 vs. 119.44±3.56, ROS (RFU): 92.30±2.46 vs. 121.33±6.57, Bax: 0.89±0.01 vs. 1.10±0.01, all P < 0.01]; RFU of MMP, MPTP in chondriosome and the protein expression of Bcl-2 were all increased at corresponding time points [MMP (RFU): 99.28±3.97 vs. 74.72±3.00, MPTP (RFU): 188.82±4.44 vs. 160.01±2.04, Bcl-2: 0.52±0.02 vs. 0.30±0.02, all P < 0.01]. The high expressions of cytokines and chondriosome damage were involved in the early TBI. Early treatment with an intraperitoneally injection of hydrogen-rich water can reduce chondriosome damage and inflammation factor release, reduce the nerve cell apoptosis after TBI, and protect brain function.