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269: Bax-activation during lidocaine-induced apoptosis in human neuroblastoma cells
186
Posters
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Miscellaneous
419. Treatment of postdural puncture headache with intravenous hydrocortisone I.P. Posso1, L.M. Marzochi1, W.S. Carvalho1, O.C. Pires1, M.B. Posso2 1Medicine, Universidade De Taubate, Hospital Universitario, Taubate, Brazil, 2Nursing, Universidade Do Vale Do Paraiba, Hospital Universitario, Taubate, Brazil Background and Aims: The causes of postdural puncture headache are poorly understood and several treatments are symptomatic, most are empirical and not effective. When conservative treatments fail, an alternative that may be proposed is the blood patch, an invasive technique which is not without risk and which many patients reject. The adrenocorticotropic hormone has shown good results in patients with disabling postdural puncture headache and has been proposed as alternative to epidural blood patch in refractory cases or in epidural blood patch failure. The adrenocorticotropic hormone would act stimulating the adrenal corticoids production and increase cerebrospinal fluid. The aim of this study is to determine if intravenous hydrocortisone offer an alternative to the blood patch for control of postdural puncture headache. Methods: After approval of the Ethics Committee, four female and two male patients, physical status PI and PII, experiencing headache after spinal anesthesia given for cesarean section, hemorrhoidectomy and knee arthroscopy received one dose of hydrocortisone 1000 mg intravenous and dipyrone 1000 mg orally every 6 hours and venous hydration. The site and number of punctures, the needle diameter, the time it took to control the pain and the volume of saline solution given were recorded. Results: The mean weight was 70.76 ⫹ 29.68, the mean age 40.33 ⫹16.5. The number of punctures was 1.5 ⫹ 0.84. The mean time for remission of headache was 13.67 ⫹ 6.03 hours, ranging between 2 to 24 hours. All patients received only one dose of hydrocortisone. The dipyrone dose was repeated only 4 times and the mean hydration was 2,583.33 ⫹ 664.58. Conclusions: Incapacitating postdural puncture headache symptoms were resolved in 13.67 ⫹ 6.03 hours with intravenous hydrocortisone treatment. Due to the high incidence of incapacitating headache, hydrocortisone is useful as it provides a noninvasive therapy with good results.
269. Bax-activation during lidocaineinduced apoptosis in human neuroblastoma cells M.F. Stevens1,2, S. Braun2, H. Hermanns2, P. Lipfert2, F. Essmann3, R. Werdehausen2 1Department of Anesthesiology, Academic Medical Center, Amsterdam, The Netherlands, 2Department of Anesthesioology, University Hospital Duesseldorf, Duesseldorf, Germany, 3Institute of Molecular Medicine, University Hospital Duesseldorf, Duesseldorf, Germany Background: Local anesthetics can induce apoptosis and thereby mediate local neurotoxicity. Hence, we investigated the mechanism of lidocaine-induced neurotoxicity in a human neuronal cell model. Incorporation of activated Bax-proteins into the outer mitochondrial membrane induces release apoptotic factors from the mitochondria after activation the mitochondrial pathway of apoptosis. Methods: Human neuroblastoma cells (SHEP) were incubated (24h) with different concentrations of lidocaine with and without addition of a pancaspase-inhibitor (Q-VD). After double-staining (Annexin-V/PI) apoptosis was evaluated by flowcytometry. Neuronal cells were exposed to lidocaine (6-12h) and oligomerization of Bax forming channels in the outer mitochondrial membrane was evaluated by immunhistochemistry. Results: Lidocaine lead in neuronal cells to a dose-dependent induction of apoptosis up to 10 mM. Lidocaine-induced apoptosis could be inhibited by the pancaspase-inhibitor (Fig. 1). Furthermore, after 6-12h incubation lidocaine induced oligomerization of Bax-protein at the outer mitochondrial membrane (Fig. 2). Conclusions: The observation of Bax-oligomers at the mitochondria demonstrates the activation of the mitochondrial pathway during lidocaine-induced neuroapoptosis. Although lidocaine induces release of apoptotic factors from the mitochondria, its apoptosis-inducing action can be oppressed by a caspase-inhibitor further down the apoptotic cascade.
Posters
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Miscellaneous
419. Treatment of postdural puncture headache with intravenous hydrocortisone I.P. Posso1, L.M. Marzochi1, W.S. Carvalho1, O.C. Pires1, M.B. Posso2 1Medicine, Universidade De Taubate, Hospital Universitario, Taubate, Brazil, 2Nursing, Universidade Do Vale Do Paraiba, Hospital Universitario, Taubate, Brazil Background and Aims: The causes of postdural puncture headache are poorly understood and several treatments are symptomatic, most are empirical and not effective. When conservative treatments fail, an alternative that may be proposed is the blood patch, an invasive technique which is not without risk and which many patients reject. The adrenocorticotropic hormone has shown good results in patients with disabling postdural puncture headache and has been proposed as alternative to epidural blood patch in refractory cases or in epidural blood patch failure. The adrenocorticotropic hormone would act stimulating the adrenal corticoids production and increase cerebrospinal fluid. The aim of this study is to determine if intravenous hydrocortisone offer an alternative to the blood patch for control of postdural puncture headache. Methods: After approval of the Ethics Committee, four female and two male patients, physical status PI and PII, experiencing headache after spinal anesthesia given for cesarean section, hemorrhoidectomy and knee arthroscopy received one dose of hydrocortisone 1000 mg intravenous and dipyrone 1000 mg orally every 6 hours and venous hydration. The site and number of punctures, the needle diameter, the time it took to control the pain and the volume of saline solution given were recorded. Results: The mean weight was 70.76 ⫹ 29.68, the mean age 40.33 ⫹16.5. The number of punctures was 1.5 ⫹ 0.84. The mean time for remission of headache was 13.67 ⫹ 6.03 hours, ranging between 2 to 24 hours. All patients received only one dose of hydrocortisone. The dipyrone dose was repeated only 4 times and the mean hydration was 2,583.33 ⫹ 664.58. Conclusions: Incapacitating postdural puncture headache symptoms were resolved in 13.67 ⫹ 6.03 hours with intravenous hydrocortisone treatment. Due to the high incidence of incapacitating headache, hydrocortisone is useful as it provides a noninvasive therapy with good results.
269. Bax-activation during lidocaineinduced apoptosis in human neuroblastoma cells M.F. Stevens1,2, S. Braun2, H. Hermanns2, P. Lipfert2, F. Essmann3, R. Werdehausen2 1Department of Anesthesiology, Academic Medical Center, Amsterdam, The Netherlands, 2Department of Anesthesioology, University Hospital Duesseldorf, Duesseldorf, Germany, 3Institute of Molecular Medicine, University Hospital Duesseldorf, Duesseldorf, Germany Background: Local anesthetics can induce apoptosis and thereby mediate local neurotoxicity. Hence, we investigated the mechanism of lidocaine-induced neurotoxicity in a human neuronal cell model. Incorporation of activated Bax-proteins into the outer mitochondrial membrane induces release apoptotic factors from the mitochondria after activation the mitochondrial pathway of apoptosis. Methods: Human neuroblastoma cells (SHEP) were incubated (24h) with different concentrations of lidocaine with and without addition of a pancaspase-inhibitor (Q-VD). After double-staining (Annexin-V/PI) apoptosis was evaluated by flowcytometry. Neuronal cells were exposed to lidocaine (6-12h) and oligomerization of Bax forming channels in the outer mitochondrial membrane was evaluated by immunhistochemistry. Results: Lidocaine lead in neuronal cells to a dose-dependent induction of apoptosis up to 10 mM. Lidocaine-induced apoptosis could be inhibited by the pancaspase-inhibitor (Fig. 1). Furthermore, after 6-12h incubation lidocaine induced oligomerization of Bax-protein at the outer mitochondrial membrane (Fig. 2). Conclusions: The observation of Bax-oligomers at the mitochondria demonstrates the activation of the mitochondrial pathway during lidocaine-induced neuroapoptosis. Although lidocaine induces release of apoptotic factors from the mitochondria, its apoptosis-inducing action can be oppressed by a caspase-inhibitor further down the apoptotic cascade.