[Influence of acupuncture plus moxibustion on changes of synaptic structure and expression of synaptic skeleton related genes and proteins in prefrontal cortex of heroin re-addicted rats].

OBJECTIVE: To observe the effect of acupuncture plus moxibustion on the synaptic ultrastructure and expression of synaptic skeleton related proteins in the prefrontal cortex (PFC) of heroin re-addicted rats, so as to reveal its mechanisms underlying improvement of heroin addiction. METHODS: Twenty-four Wister rats (half male and half female) were randomly divi-ded into normal control, model and acupuncture groups (n＝8 in each group). The heroin re-addicted model was established by muscular injection of heroin into the hind limbs for 8 days (incremental 0.8-3.6 mg, once daily for 6 days, and twice daily for 2 days), followed by conventional breeding for 5 days (detoxification), the procedure (addition－detoxification) was repeated 3 cycles. For rats of the acupuncture group, "Baihui" (GV20) was needled with filiform needles which were retained for 30 min, and moxibustion was then applied to bilateral "Shenshu" (BL23) for 30 min. The treatment was conducted once daily during the deto-xification. On the 39th day of experiment, the bilateral prefrontal cortex tissues were sampled for examining the ultrastructure by using transmission electron microscope (TEM) after fixative solution immersion and for determining the expression of genes and proteins of activity-regulated cytoskeleton-associated protein (Arc), microtubule-asso-ciated protein-2 (MAP-2) and microtubule-associated protein Tau (Tau) with quantitative real-time PCR and Western blot, respectively. RESULTS: After modeling, the expression levels of Arc mRNA and protein were significantly up-regulated, and those of MAP-2 and Tau mRNA and proteins ob-viously down-regulated in the model group relevant to the normal control group (P<0.05). Following the intervention, the up-regulated Arc protein and mRNA and the down-regulated MAP-2 and Tau were obviously reversed relevant to the model group (P<0.05). Outcomes of TEM showed unclear pre- and post-membranes of the synapses, narrowing of the synaptic gap and non-uniform of the density of the thickened dense plaque after modeling, which was relatively milder in the acupuncture group. CONCLUSION: Acupuncture plus moxibustion can improve changes of synaptic ultrastructure in heroin re-addicted rats, which may be related to their effect in regulating the expression of some synaptic skeleton proteins and genes.

Regulation of the endoplasmic reticulum stress response and neuroprotective effects of acupuncture on brain injury caused by heroin addiction.

OBJECTIVES: To evaluate regulation of the endoplasmic reticulum stress (ERS) response by acupuncture and to investigate its neuroprotective effect on brain injury caused by heroin addiction. METHODS: A total of 48 male Sprague-Dawley rats were randomly divided into a healthy control group (Control), an untreated heroin exposed group (Heroin) and a heroin exposed group receiving electroacupuncture (EA) treatment at GV14 and GV20 (Heroin+acupuncture) with n=16 rats per group. A rat model of heroin addiction was established by intramuscular injection of incremental doses of heroin for 8 consecutive days. A rat model of heroin relapse was established according to the exposure (addiction) → detoxification method. Apoptotic changes in nerve cells in the hippocampus and ventral tegmental area (VTA) were evaluated in each group of rats using terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay. PERK, eIF2a, CHOP, IRE1 and JNK gene expression and protein expression were measured using quantitative real-time PCR (RT-qPCR) assay and immunohistochemical assay, respectively. RESULTS: The total number of positive nerve cells in the hippocampus and VTA was significantly lower in the Heroin+acupuncture group than in the Heroin group (p<0.01). Compared with the Heroin group, mRNA and protein expression of PERK, eIF2a, CHOP, IRE1 and JNK in the hippocampus and VTA were significantly downregulated in the Heroin+acupuncture group (p<0.05). CONCLUSION: The acupuncture-regulated ERS response appears to mediate the neuroprotective effect of acupuncture in heroin-addicted rats with brain injury. Inhibition of CHOP and JNK upregulation and reduction of nerve cell apoptosis may be the main mechanisms underlying the effects of acupuncture on heroin addiction-induced brain injury.

Acupuncture regulates the unfolded protein response and inhibits apoptosis in a rat model of heroin relapse.

OBJECT: To explore the unfolded protein response (UPR) in the hippocampus of rats undergoing heroin relapse and the mechanisms underlying the acupuncture-mediated inhibition of brain damage caused by heroin relapse. METHODS: 60 Sprague-Dawley rats (30 females and 30 males) were randomly divided into four groups: Control group, Heroin group, Heroin+acupuncture group, and Heroin+methadone group (n=15 each). In the latter three groups, a model of heroin addiction was established by successive increments of intramuscular heroin injections for 8 days, according to the exposure (addiction)→detoxification method. A UPR RT2 Profiler PCR array was used to screen for differentially expressed genes in the hippocampus. Apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) staining. The protein expression levels of the following three differentially expressed genes were detected by Western blot to validate the results of the PCR array: heat shock protein (HSP)70, HSP105, and valosin-containing protein (Vcp). RESULTS: The UPR RT2 Profiler PCR Array detection results indicated that acupuncture increased the expression levels of the molecular chaperones HSP70, HSP105, and Vcp. The degree of neuronal apoptosis in the hippocampus of rats in the Heroin+acupuncture and Heroin+methadone groups was significantly reduced compared with the untreated Heroin group (p<0.01). Protein expression of HSP70, HSP105, and Vcp in the Heroin+acupuncture and Heroin+methadone groups was significantly higher than the Heroin group (p<0.01). CONCLUSIONS: The positive effects of acupuncture on brain damage caused by heroin may be closely related to up-regulation of HSP70, HSP105, and Vcp, and reduced apoptosis.

[Effect of manual acupuncture stimulation of "Baihui" (GV 20) and "Dazhui" (GV 14) on contents of 5-HT, dopamine and ACh and expression of 5-HT mRNA, DA mRNA and AChE mRNA in the hippocampus in methamphetamine addiction rats].

OBJECTIVE: To observe the effect of manual acupuncture stimulation on changes of hippocampal monoamine neurotransmitter levels and expression of 5-hydorxytryptamine (5-HT) mRNA, dopamine (DA) mRNA and acetylcholine esterase (AChE) mRNA in methamphetamine addiction rats, so as to explore its mechanism underlying improvement of drug addiction. METHODS: SD rats were randomly divided into normal control, model and manual acupuncture groups (n=10 in each group). Drug addiction model was established by i.p. of methamphetamine (5 mg/kg), once a day for 15 days. Manual acupuncture stimu- lation was applied to "Baihui" (GV 20) and "Dazhui" (GV 14) once daily for 10 days. The contents of hippocampal 5-HT, DA, acetylcholine (ACh), AChE were measured by ELISA. The expressive Ilieels of hippocampal 5-HT mRNA, DA mRNA and AChE mRNA were determined by fluorescence quantitative RT-POR. RESULTS: In comparison with the normal control group, the con- tents of 5-HT, DA, ACh and AChE and the expression levels of 5-HT mRNA, DA mRNA and AChE mRNA were significantly increased in the model group (P<0.01, P<0.05). After acupuncture intervention, the levels of the above-mentioned 7 indexes were uniformly and significantly down-regulated in the manual acupuncture group (P<0.01, P<0.05). CONCLUSION: Manual acupuncture stimulation of GV 20 and GV 14 can adjust methamphetamine addiction-induced changes of some hippocampal monoa- mine neurotransmitters and expression levels of 5-HT, DA and AChE genes.

[Survey of studies on drug abstinence with acupuncture in recent 10 years].

The effect of acupuncture on substance withdrawl syndromes and craving relapse prevention of the recent 10 years were reviewed as well as its mechanism. The therapeutic effect and the possible mechanism were analyzed on the basis. From the three aspects of anti protracted abstinence symptoms, craving relapse prevention and mechanism of acupuncture, the development tendency and the prospect of application on drug withdrawl with acupuncture were expected. And it is proposed that clinical observation of acupuncture intervention on craving should be developed, the mechanism of acupuncture impact on cognitive behavior, blocking study and memory processing related to drug addiction should be explored, so as to further give play to the advantages of acupuncture on anti-drug addiction.

[Influence of acupuncture and moxibustion on conditional position preference and prefrontal cortical ultrastructure in heroin re-addicted rats].

OBJECTIVE: To observe the effect of acupuncture and moxibustion on the conditional position preference (CPP) and prefrontal cortical ultrastructure in heroin re-addicted rats. METHODS: Thirty-two Wistar rats were randomly divided into normal, model, medication and acu-moxibustion (acu-moxi) groups, with 8 cases in each. Re-addiction model was established by repeated intramuscular injection of heroin into the hindlimbs. Rats of medication group were treated with intragastric administration of Methadone during detoxification. For rats in acu-moxi group, acupuncture needle was inserted into "Baihui" (GV 20) and moxibustion treatment was used to bilateral "Shenshu" (BL 23) for 30 min during detoxification. Rats in normal group were given with intramuscular injection of normal saline during addiction. CPP tests were conducted once daily in a spatial place preference box for 8 days. On the 39th day of the experiment, the rats under anesthesia were killed for sampling the prefrontal cortex (PFC) tissue on an ice plate, then, the ultrastructure of the neurons was observed by using a transmission electron microscope. RESULTS: After modeling, the rats' staying duration in the dark-box was shortened obviously (P < 0. 1). After the treatment, the staying duration in dark-box in acu-moxi group was significantly longer than that in model and medication groups (P < 0.01). Compared with normal control group, electron-microscopic results indicated that in PFC tissues of model and medication groups, vacuoles and edema of cytoplasm at different degrees, enlargement of the rough endoplasmic reticulum (REPR), vacuoles or disappearance of mitochondria, reduction of ribosome in number, and widening of the nuclear intermembrance interstice were seen. While in acu-moxi group, the number of mitochondria and ribosome increased slightly, the mitochondrial cristae was clear, the number of REPR increased relatively but dilated slightly, and the intermembrance space was basically normal. CONCLUSION: Acu-moxibustion can improve heroin re-addicted rats' conditional position preference and has a protection effect on the cerebral cortical neurons. disappearance of mitochondria, reduction of ribosome in number, and widening of the nuclear intermembrance interstice were seen. While in acu-moxi group, the number of mitochondria and ribosome increased slightly, the mitochondrial cristae was clear, the number of REPR increased relatively but dilated slightly, and the intermembrance space was basically normal. CONCLUSION: Acu-moxibustion can improve heroin re-addicted rats' conditional position preference and has a protection effect on the cerebral cortical neurons.