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3.331 DYSFUNCTIONAL AUTOPHAGY CONTRIBUTES TO LPS-STIMULATED INFLAMMATION IN MACROPHAGES
Wednesday, 14 December 2011 / Parkinsonism and Related Disorders 18S2 (2012) S161–S234
mTOR, Bcl-2, Tau [pS199] and Tau [pS396] were detected using Western-blot. Results: In aged SAMP8 mice, mTOR activity signaling upregulated in the aged SAMP8 mice and neurons-SAMP8 at the tenth day. In the neurons-SAMP8 group, the levels of Tau [pS199] and Tau [pS396]) were significantly higher than the control neurons-SAMR1 group, and when pretreated with 0.5 mM rapamycin for 3days, the presentation of Tau [pS199] and Tau [pS396] all decreased significantly. Rapamycin administration could significantly increase the expression of LC3-II and Beclin 1 in neurons-SAMP8, and significantly reduced the levels of phosphorylated p70S6K at Thr389. Rapamycin could induce antophagy activity and inhibit mTOR signaling. Conclusion: mTOR signaling participate the neurodegenerative process of SAMP8 mice and rapamycin administration could protect neurons and alleviate the Tau phosphorylation of SAMP8 mice. Rapamycin may be useful for curing the cognitive decline of SAMP8 mice. 3.331 DYSFUNCTIONAL AUTOPHAGY CONTRIBUTES TO LPS-STIMULATED INFLAMMATION IN MACROPHAGES L.-D. Cao1,2 , L.-F. Hu2 , Y.-P. Yang1,2 , H.-F. Zheng1,2 , C.-F. Liu1,2 . 1 Department of Neurology, Second Affiliated Hospital of Soochow University, 2 Institute of Neuroscience, Soochow University, Suzhou, China Objective: To investigate whether autophagy was involved in lipopolysaccharide (LPS)-stimulated inflammation in macrophages. Methods: LPS was used to stimulate and induce the production of inflammatory cytokines in Raw264.7 macrophage. The expression of autophagy related proteins (LC3, P62 and Beclin 1) were determined by Western blot analysis. The levels of inflammatory cytokines such as TNF-a, ICAM-1and NO in cell-free culture supernatant were determined by ELISA and Griess Reagent, respectively. Results: The stimulation with LPS markedly increased the level of autophagy related proteins of LC3, P62 and Beclin1 in Raw 264.7 macrophage. Pretreatment with 3-MA, an autophagy inhibitor significantly decreased the levels of autophagy related proteins and also suppressed the production of inflammation cytokines including TNF-a, ICAM-1 and NO induced by LPS. But, pretreatment with Rapamycin, an autophagy activator significantly increased the levels of autophagy related proteins and enhanced the LPS-stimulated expression of inflammation cytokines. Conclusion: LPS can trigger the dysfunction of autophagy and inflammatory responses in macrophage. Autophagy inhibition, at early stage, can alleviate the accumulation of P62 and inflammatory cytokines in LPS-stimulated macrophage. 3.333 DETERMINATION OF COPPER AS TRACE METAL IN BLOOD SERUM OF MALARIAL PATIENTS BY ATOMIC ABSORPTION SPECTROSCOPY S. Baloch1 , G.S. Gachal1 , S.A. Memon2 , M. Baloch2 . Chemistry, Zoology, Biochemistry, 1 Department of Zoology, 2 Dr. M.A. Kazi, Institute of Chemistry, University of Sindh, Jamshoro, Pakistan Malaria is one of the most serious tropical diseases in the world which has been threatens the humans by health risk from several generations. It is very widespread disease covering indefinite boundaries of Europe, North America, South America, Asia and Africa. It is also a severe public health problem in Pakistan due to poor Hygienic condition, malnutrition borne non-defensive immunity system. Copper is a very stimulating mineral to the nerves and nervous system. Copper can increase the production of neurotransmitter, and these ascending levels can burst forth the psychological
S227
disequilibrium which could be seen as mood swings, depression, mental agitation, feeling over-stimulated, restlessness, anxiety, insomnia and a racing mind with too many thoughts are all hallmarks of elevated Copper toxicity. The Plasmodium parasite invades an erythrocyte host cell containing copper. In the present study, copper level was determined in the blood serum of Malarial Patients (n = 12) with comparison to control subjects (n = 12). Metal copper was determined using Atomic Absorption Spectroscopy (AAS, Model Varian A-20) The blood serum level of Copper determined to be 2.6917 ppm, is higher as compared to the 2.045 ppm in normal subjects, with p < 0.001. 3.334 BETAHISTINE IS EFFECTIVE IN CONTROLLING SPASMS AND IMPROVES LEARNING AND MNEMONIC ABILITY IN NMDA-INDUCING INFANTILE SPASMS RAT MODEL F. Jia1 , C. Niu2 , H. Jiang1 , H. Li1 . 1 First Hospital of Jilin University, 2 Japan-China Union Hospital of Jilin University, Changchun, China Objective: To investigate the effects of betahistine on spasms control and learning and memory in NMDA-inducing infantile spasms rat model. Methods: Eighty-eight 12-day-old Wistar rats were divided into saline control group, NMDA model group and BH treatment group. Rats in saline control group were intraperitoneally injected with saline; rats of NMDA model group were intraperitoneally injected with NMDA; Rats of betahistine group were intragastrically administrated with betahistine dihydrochloride then followed by NMDA injection. Pups of betahistine group were divided into high and low dosage groups, and the rats were administered with betahitine dihydrochloride at the dosages as 300 mg/kg and 150 mg/kg, respectively. The treatment was carried out for continous 14 days. Behaviors of each rat were monitored using a video camera for two hours following the drug administration. Latency to tail-flicking and emprosthonous were recorded. Learning and mnemonic ability was assessed by Morris water test. Histamine content of prefrontal cortex and hippocampus was evaluted by HPLC technique. Histamine H3 receptor expression was examined by immunohistochemistry. Result: Incidence of tail-flicking and emprothonus was significantly lower in the betahistine groups when compared with NMDA group. Latencies to tail-flicking and emprothonus were significantly longer in betahistine groups than those in the NMDA model group. Betahistine shortened the escape time in a dose-dependent manner when compared with NMDA group. Betahistine increased histamine content and H3 receptor expression in the brain in a dosagedependent manner. Conclusion: Betahistine is effective in controlling spasms and improves the learning and mnemonic ability in the NMDA-inducing infantile spasms rat model. 3.335 DUAL-RESONANCE GENERATED BY SK-CURRENT AND H-CURRENT AT HYPERPOLARIZED MEMBRANE POTENTIALS AT THETA FREQUENCY IN RAT SNC NEURONS W.-N. Xue, Y. Wang, Z.-Q. Yan, J.-L. Zhu, S.-M. He, X.-L. Wang, G.-D. Gao. Neurosurgery, Tangdu Hospital, Fourth Military Medical University, Xi’an, China Objectives: Oscillation activities are the feature of neural network, and correlated to different physiological states. The theta (q) oscillation (2–7 Hz) has been reported in basal ganglia and the intrinsic resonance properties of individual neurons have provided a basis for this network oscillation. The basal ganglia neurons receive comprehensive modulation arising from dopaminergic neurons located in substantia nigra pars compacta (SNc), but how the oscillation is regulated in SNc neurons remains to be poorly understood.
mTOR, Bcl-2, Tau [pS199] and Tau [pS396] were detected using Western-blot. Results: In aged SAMP8 mice, mTOR activity signaling upregulated in the aged SAMP8 mice and neurons-SAMP8 at the tenth day. In the neurons-SAMP8 group, the levels of Tau [pS199] and Tau [pS396]) were significantly higher than the control neurons-SAMR1 group, and when pretreated with 0.5 mM rapamycin for 3days, the presentation of Tau [pS199] and Tau [pS396] all decreased significantly. Rapamycin administration could significantly increase the expression of LC3-II and Beclin 1 in neurons-SAMP8, and significantly reduced the levels of phosphorylated p70S6K at Thr389. Rapamycin could induce antophagy activity and inhibit mTOR signaling. Conclusion: mTOR signaling participate the neurodegenerative process of SAMP8 mice and rapamycin administration could protect neurons and alleviate the Tau phosphorylation of SAMP8 mice. Rapamycin may be useful for curing the cognitive decline of SAMP8 mice. 3.331 DYSFUNCTIONAL AUTOPHAGY CONTRIBUTES TO LPS-STIMULATED INFLAMMATION IN MACROPHAGES L.-D. Cao1,2 , L.-F. Hu2 , Y.-P. Yang1,2 , H.-F. Zheng1,2 , C.-F. Liu1,2 . 1 Department of Neurology, Second Affiliated Hospital of Soochow University, 2 Institute of Neuroscience, Soochow University, Suzhou, China Objective: To investigate whether autophagy was involved in lipopolysaccharide (LPS)-stimulated inflammation in macrophages. Methods: LPS was used to stimulate and induce the production of inflammatory cytokines in Raw264.7 macrophage. The expression of autophagy related proteins (LC3, P62 and Beclin 1) were determined by Western blot analysis. The levels of inflammatory cytokines such as TNF-a, ICAM-1and NO in cell-free culture supernatant were determined by ELISA and Griess Reagent, respectively. Results: The stimulation with LPS markedly increased the level of autophagy related proteins of LC3, P62 and Beclin1 in Raw 264.7 macrophage. Pretreatment with 3-MA, an autophagy inhibitor significantly decreased the levels of autophagy related proteins and also suppressed the production of inflammation cytokines including TNF-a, ICAM-1 and NO induced by LPS. But, pretreatment with Rapamycin, an autophagy activator significantly increased the levels of autophagy related proteins and enhanced the LPS-stimulated expression of inflammation cytokines. Conclusion: LPS can trigger the dysfunction of autophagy and inflammatory responses in macrophage. Autophagy inhibition, at early stage, can alleviate the accumulation of P62 and inflammatory cytokines in LPS-stimulated macrophage. 3.333 DETERMINATION OF COPPER AS TRACE METAL IN BLOOD SERUM OF MALARIAL PATIENTS BY ATOMIC ABSORPTION SPECTROSCOPY S. Baloch1 , G.S. Gachal1 , S.A. Memon2 , M. Baloch2 . Chemistry, Zoology, Biochemistry, 1 Department of Zoology, 2 Dr. M.A. Kazi, Institute of Chemistry, University of Sindh, Jamshoro, Pakistan Malaria is one of the most serious tropical diseases in the world which has been threatens the humans by health risk from several generations. It is very widespread disease covering indefinite boundaries of Europe, North America, South America, Asia and Africa. It is also a severe public health problem in Pakistan due to poor Hygienic condition, malnutrition borne non-defensive immunity system. Copper is a very stimulating mineral to the nerves and nervous system. Copper can increase the production of neurotransmitter, and these ascending levels can burst forth the psychological
S227
disequilibrium which could be seen as mood swings, depression, mental agitation, feeling over-stimulated, restlessness, anxiety, insomnia and a racing mind with too many thoughts are all hallmarks of elevated Copper toxicity. The Plasmodium parasite invades an erythrocyte host cell containing copper. In the present study, copper level was determined in the blood serum of Malarial Patients (n = 12) with comparison to control subjects (n = 12). Metal copper was determined using Atomic Absorption Spectroscopy (AAS, Model Varian A-20) The blood serum level of Copper determined to be 2.6917 ppm, is higher as compared to the 2.045 ppm in normal subjects, with p < 0.001. 3.334 BETAHISTINE IS EFFECTIVE IN CONTROLLING SPASMS AND IMPROVES LEARNING AND MNEMONIC ABILITY IN NMDA-INDUCING INFANTILE SPASMS RAT MODEL F. Jia1 , C. Niu2 , H. Jiang1 , H. Li1 . 1 First Hospital of Jilin University, 2 Japan-China Union Hospital of Jilin University, Changchun, China Objective: To investigate the effects of betahistine on spasms control and learning and memory in NMDA-inducing infantile spasms rat model. Methods: Eighty-eight 12-day-old Wistar rats were divided into saline control group, NMDA model group and BH treatment group. Rats in saline control group were intraperitoneally injected with saline; rats of NMDA model group were intraperitoneally injected with NMDA; Rats of betahistine group were intragastrically administrated with betahistine dihydrochloride then followed by NMDA injection. Pups of betahistine group were divided into high and low dosage groups, and the rats were administered with betahitine dihydrochloride at the dosages as 300 mg/kg and 150 mg/kg, respectively. The treatment was carried out for continous 14 days. Behaviors of each rat were monitored using a video camera for two hours following the drug administration. Latency to tail-flicking and emprosthonous were recorded. Learning and mnemonic ability was assessed by Morris water test. Histamine content of prefrontal cortex and hippocampus was evaluted by HPLC technique. Histamine H3 receptor expression was examined by immunohistochemistry. Result: Incidence of tail-flicking and emprothonus was significantly lower in the betahistine groups when compared with NMDA group. Latencies to tail-flicking and emprothonus were significantly longer in betahistine groups than those in the NMDA model group. Betahistine shortened the escape time in a dose-dependent manner when compared with NMDA group. Betahistine increased histamine content and H3 receptor expression in the brain in a dosagedependent manner. Conclusion: Betahistine is effective in controlling spasms and improves the learning and mnemonic ability in the NMDA-inducing infantile spasms rat model. 3.335 DUAL-RESONANCE GENERATED BY SK-CURRENT AND H-CURRENT AT HYPERPOLARIZED MEMBRANE POTENTIALS AT THETA FREQUENCY IN RAT SNC NEURONS W.-N. Xue, Y. Wang, Z.-Q. Yan, J.-L. Zhu, S.-M. He, X.-L. Wang, G.-D. Gao. Neurosurgery, Tangdu Hospital, Fourth Military Medical University, Xi’an, China Objectives: Oscillation activities are the feature of neural network, and correlated to different physiological states. The theta (q) oscillation (2–7 Hz) has been reported in basal ganglia and the intrinsic resonance properties of individual neurons have provided a basis for this network oscillation. The basal ganglia neurons receive comprehensive modulation arising from dopaminergic neurons located in substantia nigra pars compacta (SNc), but how the oscillation is regulated in SNc neurons remains to be poorly understood.