Effect of electroacupuncture intervention on the spinal cord PPIA/NF-κB signaling pathway in mice with amyotrophic lateral sclerosis. / ç”µé’ˆå¯¹è‚ŒèŽç¼©ä¾§ç´¢ç¡¬åŒ–ç—‡å°é¼ è„Šé«“PPIA/NF-κB信号通路的影响.

OBJECTIVES: To observe the effects of electroacupuncture (EA) on motor function, expression of extracellular cyclophile A(PPIA) and PPIA/nuclear factor-κB (NF-κB) signaling pathway in spinal cord of amyotrophic la-teral sclerosis (ALS) mice, so as to explore the mechanism of EA intervention in regulating extracellular PPIA on neuroinflammation in ALS mice. METHODS: Thirty ALS-SOD1G93A mice with hSOD1-G93A gene were randomly divided into model, EA and Riluzole groups , with 10 mice in each group, and other 10 ALS-SOD1G93A negative mice were used as the blank group. EA was applied to bilateral "Yanglingquan"(GB34) and "Zusanli"(ST36) for 20 min once daily, 5 days a week for 2 weeks. In the Riluzole group, riluzole solution (30 mg·kg-1·d-1) was administrated intragastrically, and the treatment time was the same as that in the EA group.Rotating rod experiment and open field experiment were used to evaluate the changes in motor function of mice .The morphology of motor neurons in the anterior horn of spinal cord was observed by HE staining.The relative protein expression levels of PPIA, TDP-43 and NF-κB in the spinal cord were detected by Western blot.The positive expression level of TDP-43 in the spinal cord was detected by immunohistochemistry. The positive expression level of PPIA in spinal cord was marked by immunofluorescence. Serum PPIA content was determined by ELISA. RESULTS: Compared with the blank group, the time of rod dropping and the total distance of open field movement in the model group were shortened (P<0.01), the number of motor neurons in the anterior horn of the spinal cord was reduced, the cell morphology was incomplete, the cell body was atrophied, the protein expression and positive expression of TDP-43 were increased (P<0.01), the protein expressions of PPIA and NF-κB in the spinal cord were increased(P<0.01), the serum content of PPIA and immunofluorescence expression of PPIA in spinal cord were increased (P<0.01). Compared with the model group, the time of rod dropping and the total distance of open field movement of mice in the EA group and the Riluzole group were prolonged (P<0.05, P<0.01), and the injury of motor neuron in the anterior horn of the spinal cord was decreased, the protein expression and positive expression of TDP-43 in the spinal cord were decreased (P<0.05, P<0.01)；the relative expression levels of PPIA and NF-κB proteins were decreased (P<0.05, P<0.01), and the content of PPIA in serum and the immunofluorescence expression of PPIA in the spinal cord were decreased (P<0.05, P<0.01) in the EA group；the relative protein expression of NF-κB and fluorescence expression of PPIA in spinal cord of mice in the Riluzole group were decreased (P<0.05). CONCLUSIONS: EA intervention can improve motor function in ALS mice, and its mechanism may be related to the inhibition of PPIA/NF-κB signaling pathway by EA to alleviating neuroinflammatory response.

[Effect of electroacupuncture on hippocampal synaptic plasticity and complement dependent memory impairment in Parkinson's disease dementia mice].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Xiusanzhen" ï¼»bilateral "Yingxiang"(LI20)+"Yintang"(GV24+)ï¼½ on synaptophysin (SYN), postsynaptic density protein-95 (PSD-95), Iba-1+ CD68+ microglia and complement C related protein expression of hippocampus in Parkinson's disease dementia (PDD) mice, so as to explore its mechanism in improving memory impairment of PDD. METHODS: Male C57BL/6 mice were randomly divided into control, sham operation, model and EA groups, with 10 mice in each group. The PDD model was established by injecting 6-OHDA into the medial forebrain tract. EA (2 Hz, 1 mA) was applied to unilateral LI20 and GV29 for 20 min once daily for consecutive 14 days. Morris water maze and new object recognition test were used to evaluate the learning and memory ability. Western blot was used to detect the expression of SYN and PSD-95 proteins in hippocampus. Immunofluorescence was used to label Iba-1+ CD68+ microglia and C1q positive cells in hippocampal CA1 region. The content of C3 protein in hippocampus was detected by ELISA. RESULTS: Compared with the control group, there was no statistical significance in all the observed indexes in the sham operation group. Compared with the sham operation group, the average escape latency (AEL) prolonged significantly (P<0.01), the target platform crossing times (TPCT) and new object recognition index (NORI) decreased remarkably (P<0.01); the expressions of SYN and PSD-95 proteins in hippocampal CA1 region were significantly decreased (P<0.01); the rate of Iba-1+CD68+ microglia, the rate of C1q positive cells and the content of C3 protein were significantly increased (P<0.01) in the model group. In comparison with the model group, the AEL was shortened (P<0.01), the TPCT and NORI were increased (P<0.05) remarkably; the expressions of SYN and PSD-95 proteins in hippocampal CA1 region were increased (P<0.01, P<0.05); the rate of Iba-1+ CD68+ microglia, the rate of C1q positive cells and the content of C3 protein were significantly decreased (P<0.01) in the EA group. CONCLUSION: "Xiusanzhen" can alleviate the learning and memory impairment of PDD model mice, and improve the synaptic plasticity of hippocampal CA1 area. The mechanism may be related to the reduction of C1q and C3 deposition in hippocampal CA1 region and the reduction of microglia phagocytosis.

[ZHANG Wei-hua's experience in clinical treatment of somatalgia with "point-zone-strip stimulation technique" of acupuncture].

The clinical experience in treatment of somatalgia with "point-zone-strip stimulation technique" of acupuncture of professor ZHANG Wei-hua was summarized. Professor ZHANG integrates the theories of the cutaneous region of meridian, biaoben, "taking tender point as acupoint" and local holography as a whole in treatment of somatalgia. The "point-stimulation technique" of acupuncture (subcutaneous needling technique) is adopted for the painful site less than 3 cm in diameter, the "zone-stimulation technique" (surrounding needling technique) is for the site larger than 3 cm in diameter and the "strip-stimulation technique" (cubit-tibia needling technique) is for various acute and chronic somatalgia. These three needling techniques are applicable singly or in combination in clinical practice.

Electroacupuncture inhibition of hyperalgesia in rats with adjuvant arthritis: involvement of cannabinoid receptor 1 and dopamine receptor subtypes in striatum.

Electroacupuncture (EA) has been regarded as an alternative treatment for inflammatory pain for several decades. However, the molecular mechanisms underlying the antinociceptive effect of EA have not been thoroughly clarified. Previous studies have shown that cannabinoid CB1 receptors are related to pain relief. Accumulating evidence has shown that the CB1 and dopamine systems sometimes interact and may operate synergistically in rat striatum. To our knowledge, dopamine D1/D2 receptors are involved in EA analgesia. In this study, we found that repeated EA at Zusanli (ST36) and Kunlun (BL60) acupoints resulted in marked improvements in thermal hyperalgesia. Both western blot assays and FQ-PCR analysis results showed that the levels of CB1 expression in the repeated-EA group were much higher than those in any other group (P = 0.001). The CB1-selective antagonist AM251 inhibited the effects of repeated EA by attenuating the increases in CB1 expression. The two kinds of dopamine receptors imparted different actions on the EA-induced CB1 upregulation in AA rat model. These results suggested that the strong activation of the CB1 receptor after repeated EA resulted in the concomitant phenomenon of the upregulation of D1 and D2 levels of gene expression.

[Effects of repeated electroacupuncture on gene expression of cannabinoid receptor-1 and dopamine 1 receptor in nucleus accumbens-caudate nucleus region in inflammatory-pain rats].

OBJECTIVE: To observe the effect of repeated electroacupuncture (EA) on the expression of cannabinoid receptor-1 (CB 1) mrRNA and dopamine 1 receptor (D 1) mRNA in Nucleus Accumbens (NAC)-Caudate Nucleus (CN) region in inflammatory-pain rats, so as to study its underlying mechanism in analgesia. METHODS: A total of 30 SD rats were randomized into normal control, model, EA, EA+ AM 251 and WIN 55212-2 groups, with 6 cases in each group. EA (2 Hz/100 Hz, 1 -3 mA) was applied to "Zusanli"(ST 36) and "Kunlun"(BL 60) for 30 min, once every other day, and 4 sessions all together. Arthritis model was established by injection of Freund's complete adjuvant 0.05 mL in the rat's left ankle. Thermal pain threshold (paw withdrawal latency, PWL) was detected before and after modeling and after repeated EA and/or intraperitoneal injection of AM 251(an inverse antagonist at the CB 1 cannabinoid receptor, 0. 1 mg/100 g) and WIN 55212-2 (a potent cannabinoid receptor agonist, 0. 2 mg/100 g). The expression of CB 1 receptor mRNA and D 1 receptor mRNA in the NAC-CN region was measured by real time fluorescence quantitative-polymerase chain reaction. RESULTS: Compared with the control group, the pain threshold values of the model group was decreased significantly (P<0.01). In comparison with the model group, the pain threshold values of the EA group and WIN 55212-2 group were increased considerably on day 10 (P<0. 01). No significant differences were found between the EA+ AM 251 and model groups and between the EA and WIN 55212-2 groups in PWL after the treatment (P>0.05). Compared with the control group, both CB 1 R mRNA and D 1 R mRNA expression levels in the model group were increased slightly, while in comparison with the model group and EA+ AM 251 group, CB 1 R mRNA and D 1 R mRNA expression levels in the EA group and WIN 55212-2 group were upregulated obviously. No significant differences were found between the EA + AM 251 and model groups and between the EA and WIN 55212-2 groups in CB 1 R mRNA and D 1 R mRNA expression levels.