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Objective To explore the influence of IFN-? on anti-proliferation and apoptosis of the Caspase-8 silenced neuroblastoma cell line induced by doxorubicin and the influence mechanism.Methods MTT and flow cytometric analysis were used to detect the survival and apoptosis rates before and after the combined treatment of IFN-? and doxorubicin.Immunohistochemical staining was used to detect the expression of caspase-8 protein before and after the treatment of IFN-?.Results The survival rates of doxorubicin(0.03,0.10,0.30?g/ml)used alone for 24h were(95.62?13.03)%,(82.62?7.94)%,(64.84?9.19)%,IFN-?(10,100,1000U/ml)used alone for 48h were(98.37%?11.25)%,(97.15?5.36)%,(98.84?7.41)%.The survival rates of IFN-?(1000U/ml)combined with different concentration of doxorubicin(0.03,0.10,0.30?g/ml)were significantly lower than that of doxorubicin(0.03,0.10,0.30?g/ml)alone.The apoptosis rate of doxorubicin(0.30?g/ml)alone was(22.00?6.55)%,IFN-?(1000U/ml)alone was (8.22?4.00)%,whereas the apoptosis rate of the combined treatment of IFN-? and doxorubicin wassignificantly higher than that of doxorubicin alone;Immunohistochemical staining showed that caspase-8 protein was negative in SH-SY5Y cell line,whereas it become positiveafter the treatment of IFN-?(1000U/ml)for 48h.Conclusion Treating human neuroblastoma cell line SH-SY5Y cell with doxorubicin may induce anti-proliferation and apoptosis.Combined treatment with IFN-? may significantly increase this effect.The mechanism may be realized by upregulating the expression of caspase-8 protein.
ABSTRACT
BACKGROUND: The active ingredients of Chinese herbs, such as hongjingtian (rhodiola root, Radix Rhodiolae), srenshen (ginseng, Radix Ginseng), and danggui (Chinese angelica root, Radix Angelicae Sinensis),were found to be effective to inhibit the hypoxia injury.OBJECTIVE: To observe the protecting effect of xionggui diwan (Dripping Pills) on the hippocampal neurons from hypoxia by whole-cell recording patch-clamp technique.DESIGN: An auto-control observation on cells.SETTING: The 88 Hospital of Chinese PLA and Academy of Military Medical Science of Chinese PLA.MATERIALS: The experiment was performed during March to August 2002 in the Institute of Toxicology and Pharmacology of Academy of Military Medical Sciences of Chinese PLA. The neonatal Wistar rats, clean degree, were used. The hippocampal neuron cells out of 9 rats were cultured and divided into the control groups of before and after hypoxia-oxygenation,and xionggui diwan 1 × 10-6 mol/L and 10 × 10-6 mol/L groups. (Xionggui diwan was provided by the 88 Hospital of Chinese PLA and its active ingredient was the volatile ingredient of chuanxiong and danggui.)METHODS: While hypoxia-reoxygenation groups were not given any medicine intervention as control, the treatment of xionggui diwan 1×10-6mol/L was subdivided into xionggui diwan 1 × 10-6 mol/L + non-hypoxia and xionggui diwan 1 × 10-6 mol/L + hypoxia groups, and the treatment of xionggui diwan 10 × 10-6 mol/L was subdivided into xionggui diwan 10 × 10-6 mol/L + non-hypoxia and xionggui diwan 10 × 10-6 mol/L + hypoxia groups. The patch-clamp technique was performed to determine the Na+ and K+ currents of rat hippocampal neurons cultured in vitro before and after hypoxia-reoxygenation.MAIN OUTCOME MEASURES: The Na+ and K+ current amplitude of rat hippocampal neurons cultured in vitro.RESULTS:① The Na+ current amplitude of hippocampal neurons: After hypoxia-reoxygenation, it was significantly lower than that before [(-3.8±1.0, -10.3±0.5) nA, t=3.49, P ＜ 0.01]. ② The K+ current amplitude of hippocampal neurons: After hypoxia-reoxygenation, the activation thresholdof K+ current was changed from -40 mV to -20 mV.③The Na+ and K+ currents of hippocampal neurons before and after hypoxia-reoxygenation with xionggui diwan pretreatment: Before hypoxia-reoxygenation, the ampli tudes of Ⅰ Na and J K of hippocampal neurons at the threshold point were very similar, without significant difference. After hypoxia-reoxygenation,those amplitudes of xionggui diwan 10.0 × 10-6 mol/L group were higher than that of xiongguidiwan 1.0 × 10-6 mol/L group [(-58.5±1.9, -6.9±1.4,-3.8±1.0) nA, t=7.51,P ＜ 0.05].CONCLUSION: It was suggested that xionggui diwan propect the function of hippocampal neurons from hypoxia.
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OBJECTIVE:To observe the effect of Xionggui dripping pills on sensitivity of baroreflex in selective sinoaortic denervated rats.METHODS:The dynamic blood pressures were monitored in selective sinoaortic denervated rats in waking stait,and the changes of baroreflex senitivity(BRS)were measured by modified SU Smyth method.RESULTS:Xionggui drip_ ping pill could improve BRS in sinus denervated rats.CONCLUSION:Since Xionggui dripping pill rectifies autonomic nerve function,it can protect target organs from damage in cardiovascular diseases.