Influence of morphine on levels of type Ⅱ inhibitory guanine nucleotide binding protein in primary hippocampal neurons★
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Publisher：Quzwzb Publish Time：Tuesday, November 25, 2008
Source：Neural Regen Res,2008,3(5),465-8 |
Qinghua Wu1, Qiang Fu2, Xinhua Wang3, Jianhua Zhao4, Liwei Liu3, Shirong Tang1
1Department of Anesthesiology, the 150 Hospital of Chinese PLA, Luoyang 471031, Henan Province, China
2Department of Anesthesiology, General Hospital of Chinese PLA, Beijing 100853, China
3Department of Anesthesiology, Orient Hospital Affiliated to Shanghai Tongji University, Shanghai 200120, China
4Department of Anesthesiology, General Hospital of Jinan Military Area Command of Chinese PLA, Jinan 250031, Shandong Province, China
Qinghua Wu★, Master, Attending physician, Department of Anesthesiology, the 150 Hospital of Chinese PLA, Luoyang 471031, Henan Province, China
Wu QH, Fu Q, Wang XH, Zhao JH, Liu LW, Tang SR. Influence of morphine on levels of type Ⅱ inhibitory guanine nucleotide binding protein in primary hippocampal neurons. Neural Regen Res 2008;3(5):465-8
| Abstract
BACKGROUND: The pharmacological action of opioid drugs is related to signal transduction of inhibitory guanine nucleotide binding protein.
OBJECTIVE: To quantitatively and qualitatively analyze the influence of morphine on levels of type Ⅱ inhibitory guanine nucleotide binding protein (Gi2 protein) in primary cultured hippocampal neurons at different time points.
DESIGN, TIME AND SETTING: A randomized controlled study, which was performed at the Department of Neurobiology, Changzheng Hospital, Second Military Medical University of Chinese PLA between September 2002 and March 2004.
MATERIALS: Cerebral hippocampal neurons were obtained from newborn SD rats at 1-2 days of age. Biotin-antibody II-avidin fluorescein isothiocyanate (Avidin-FITC) was purchased from Sigma Company (USA) and the Gi2 protein polyclonal antibody from Santa Cruz Biochemistry Company (USA).
METHODS: Seven days after culture, mature hippocampal neurons were randomly divided into six groups: 4-, 8-, 16-, 24-, and 48-hour morphine groups, and a blank control group. Neurons in the morphine groups received morphine (10 μmol/L), which could cause alterations of G-protein mRNA and cAMP expression in the prefrontal cortex. Neurons in the blank control group were given the same volume of saline.
MAIN OUTCOME MEASURES: Gi2 protein levels were detected by an immunofluorescence technique, and were analyzed by the image analytic system with the use of green fluorescence intensity.
RESULTS: Gi2 protein levels in hippocampal neurons gradually decreased in the 4-, 8-, 16-, 24-, and 48-hour morphine groups. In particular, Gi2 protein levels in the 16-, 24-, and 48-hour morphine groups were significantly lower than that in the blank control group (P < 0.05-0.01).
CONCLUSION: Morphine may decrease Gi2 protein level in primary hippocampal neurons, and the decreasing trend is positively related to morphine-induced time.
Key Words: morphine; hippocampal neurons; immunofluorescence; type Ⅱ inhibitory guanine nucleotide binding protein
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