Expression of specific gene related to the heroin dependence
- VernacularTitle:与海洛因依赖有关的特异性基因表达
- Author:
Huaqing MENG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2006;10(42):182-184
- CountryChina
- Language:Chinese
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Abstract:
BACKGROUND: Heroin, which is characterized by strong liposolubility and immediate effect, can rapidly enter central nervous system through blood brain barrier; however, mechanism of its dependence is still unclear up to now. Therefore, it becomes a hot topic to investigate mechanism of molecular biology of drug dependence, especially to research changes of gene expression.OBJECTIVE: To find out the expression of specific gene related to heroin dependence so as to elucidate the mechanism of molecular biology of heroin dependence.DESIGN: Randomized controlled observation on the basis of experimental animals.SETTING: Center of Psychohygiene, the First Hospital affiliated to Chongqing Medical University.MATERIALS: Eight C57BL/6J mice were randomly divided into experimental group and control group with 4 in each group. Drug and equipments: heroin, Tripure separating agent, PolyATtract mRNA separating system Ⅲ, etc.METHODS: Heroin-dependent models in mice were established by dose-increasing hypodermical injection of heroin for 7 days. On the 7th day,80 mg/kg heroin was injected once, and 2 hours later, 5 mg/kg naloxone was injected'. Mice in control group were injected with the same volume of saline and the same dosage of naloxone. Then, mice were put in glass cage to observe jumping times and numbers of animals which had ptosis, diarrhea, wet-dog movement and trembling of anterior claws within 15 minutes.Mice were immediately sacrificed and total RNA and mRNA were extracted from the brains in experimental and control groups with Trripure separating kit, and then, suppressive subtractive hybridization was proceeded.The hybridization sample was amplified by PCR and analyzed with gel electrophoresis.MAIN OUTCOME MEASURES: Quantity of total RNA and mRNA in brain tissue and different expression of cDNA sections in brain.RESULTS: Mice in each group were involved in the final analysis. ①Results of abstinent symptom after injection of naloxone: Abstinent symptom was observed on mice in experimental group. Jumping times within 15 minutes were 9.75±1.65, which was significant difference from those in control group (P ＜ 0.01). ② Comparison of total RNA and mRNA in brain tissue and different expression of cDNA sections in brain: Quantities of total RNA and mRNA were higher in experimental group than those in control group, and difference of mRNA was significant (P ＜ 0.05). After the second suppressive subtractive hybridization, mixture was amplified with PCR and nest-like PCR to differently express cDNA sections.CONCLUSION: Withdrawal of heroin dependence may cause changes of gene expression, and there is probably an expression of the specific gene in the brain of heroin-dependent mice.
