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• W2746882640 abstract "ObjectiveTo investigate the effects of regulated necrosis induced by receptor interacting- protein 3 (RIP3) on traumatic brain injury (TBI) and its mechanisms in vitro and in vivo.MethodsHT-22 cell lines were seeded on Bioflex cell plate, and was subjected to injuries with 30 psi valve pressure, 3.5~4.5 psi gas pulse pressure for 50 ms by cell injury controller. The morphological changes of HT-22 cells were assessed by real-time digital holographic microscopy (DHM), and the expressions of RIP3, MLKL, Akt, p-Akt, mTOR, and p-mTOR were detected by Western blot assay at 6 h, 12 h, 24 h post-TBI. C57BL/6 mice were then positioned beneath the controlled cortical impactor device (eCCI) device and subjected to impact injury at 1 mm depth of penetration, for a sustained depression of 150 ms and a velocity of 5 m/s. Sham-operated mice underwent identical surgical procedures, including craniotomy, without receiving the cortical impact. Finally, the expression of targeted proteins mentioned above was detected at 12 h post-CCI by SP immunohistochemistry.ResultsCompared with the control group, HT-22 cells showed a reduction in area (from (603.63±26.13)μm2 to (193.87±10.51)μm2, P<0.01) and an increase in thickness (from (3.90±0.24)μm to (7.50±0.27)μm, P<0.01) at 6 h post-CIC. Furthermore, Western blotting assay revealed that the expression of RIP3, MLKL and Akt was significantly increased (RIP3: (1.08±0.05), MLKL: (0.99±0.03), Akt: (0.97±0.02)) compared with that of control (all P<0.01) at 6-12 h, and decreased at 24 h post-CIC; the p-Akt level was gradually increased and reached the peak level (1.07±0.03) at 24 after CIC; but the expression of mTOR and p-mTOR were higher than that of control at 6 h post-CIC (mTOR: (0.78±0.02) vs (0.42±0.02), p-mTOR: (0.95±0.02) vs (0.54±0.02); all P<0.01). Additionally, compared with sham, CCI exhibited a significantly increased number of RIP3 ((497±16) vs (324±11)), MLKL ((403±12 vs (200±8)), Akt ((367±10) vs (302±14)), p-Akt ((376±10) vs (287±17)), mTOR ((144±9) vs (57±11)) and p-mTOR-positive cells ((203±7) vs (27±8)); all P<0.01) in mouse brain samples at 12 h post- CCI, in accordance with the Western blotting assay in vitro.ConclusionRegulated necrosis induced by RIP3 may play an important role in the pathogenesis of traumatic brain injury, which supports that RIP3 may be as a potential response marker for targeted drug in TBI pharmacotherapy.Key words: Receptor-interacting protein 3; Traumatic brain injury; Regulated necrosis" @default.
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• W2746882640 date "2016-10-20" @default.
• W2746882640 modified "2023-09-22" @default.
• W2746882640 title "Role of related necrosis signal pathway mediated by receptor interacting-protein 3 in traumatic brain injury" @default.
• W2746882640 doi "https://doi.org/10.3760/cma.j.issn.1674-6554.2016.10.004" @default.
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