| Object： This study was to investigate exogenous H2S by learning from rats’neuroethology and brain cells of fine structure morphology. We studiedexogenous H2S intervention for heroin addiction in rats and its mechanism byimmunohistochemical staining diagnosis, TUNEL method and other methods toprovide reference for clinical treatment of heroin addiction.Methods:1.Forty female SD rates were randomly divided into fourgroups:control group, Heroin group, NaHS group and NaHS intervention group.2.Applying the method of gradual increase of dosage,we gave the rats ofHeroin group and NaHS intervention group subcutaneous injection ofheroin.The control group and NaHS group were given subcutaneous injectionwith the equivalent normal saline.30minutes before subcutaneous injection,according to body weight of rats, NaHS group and NaHS intervention groupwere given intraperitoneal injection of NaHS. The control group and Heroingroup were injected with the equivalent normal saline.3. Naloxone was used to replace the test of hastening dependence after modeling and intervention.The symptoms of abstention from taking heroin wereevaluated by the method of Maldonado.4. Modeling and intervention confirmed through the use of MWM in each grouprats were detected on learning and memory ability.5.After MWM test，we selected from each group8rats randomly.Then wewere done general anesthesia, perfusion, removed rats’ brain, embedded inparaffin and done paraffin sections. We used immunohistochemical method todetect GFAP and NF and do Nissl staining for the SD rats’ prefrontal cortex,nucleus accumbens and three brain regions of hippocampus to do qualitativeand semi-quantitative analysis.6. TUNEL method was used to do qualitative and semi-quantitative analysis ofapoptotic cell in four groups rats’ CA1CA2CA3of hippocampus respectively.7. Two SD rats from each group were selected randomly. Ultrastructuralchanges in the rat prefrontal cortex, nucleus accumbens, three brain regions ofhippocampus were observed by transmission electron microscopy.Result：1.Compare with control group, NaHS group, NaHS intervention groupand the heroin addicts Heroin group in Morris water maze test show that thenumber of cross-platform decreased significantly and the1st time was longer tofind the platform, the difference was statistically significant (P <0.01)；However, control group, NaHS group and NaHS intervention group have nosignificant differences between each other；The record of rats in NaHSintervention group have greatly improved over Heroin group (P <0.01).2. Nissl staining: Compare with control group, NaHS group and NaHSintervention group, the number of Nissl bodies is reduced in the nucleusaccumbens and CA1,CA2,CA3areas of hippocampus, the difference wasstatistically significant (P <0.01), While among control group, NaHS group and NaHS intervention group, the number of Nissl bodies have no significantdifference.3.Immunohistochemical detection of GFAP: Compare with the other threegroups,Heroin group is increased in the number of positive cells GFAP in thenucleus accumbens and CA1,CA2,CA3areas of hippocampus, the differencewas statistically significant (P <0.01) but the control group and NaHS groupcomparison were not significant statistically significant.4.Immunohistochemical detection of NF: Compare with the other threegroups,Heroin group is increased in the number of positive cells NF in thenucleus accumbens and CA1, CA2, CA3areas of hippocampus, the differencewas statistically significant (P <0.01) but the control group and NaHS groupcomparison were not significant statistically significant.5.Apoptotic cells in brain tissue: For heroin addicts,heroin promote cellapoptosis significantly in CA1, CA2, CA3areas of hippocampus in Heroingroup compared with control group, the difference was statistically significant(P <0.01) but the control group and NaHS group comparison were notsignificant statistically significant.6.SEM showed: In the control group nerve cells as normal size, no edema, nodemyelination phenomenon, neurofilament rules capillary wall thickness andendothelial cell loss less. Compar with the control group, nerve cells is changedin Heroin group:cytoplasmic dissolution becomes sparse, increase in thenumber of apoptotic cells. neuropil structure becomes disordered, edema,disintegration. synaptic swelling, increased disintegration open synaptic vesiclesneurofilament appeared disorder, fracture. demyelination phenomenon can beseen; capillary permeability increased significantly. Conclusion：1.Using the incremental method of subcutaneous injection of heroin addiction, ratmodel, the modeling method is stable, effective.2.Heroin can lead to the decline of learning and memory of rats, increasingcerebral astrocytes content, decreased actin protein content, number of braincells apoptosis increased, capillary, neurons and nerve cell damage, andthencause a series of changes in brain tissue.3.Exogenous H2S can inhibit heroin induced brain injury on learning andmemorydecline, increasing cerebral astrocytes content, decreased actin proteincontent,number of brain cells apoptosis increased, capillary, neuronsand nerve cell damage, improve the structure and function of brain.4.Over expression of GFAP induced by exogenous H2S decreased, NF contentincreased, the survival of neurons is beneficial to the recovery of theability of learning and memory. |
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