针刺及内源性c-Fos、c-Jun对体外培养备用背根节的影响

The Effect of Acupuncture and Endogenous c-Fos,c-Jun on Regeneration of Neuronal Dendrite of Spared DRG in vitro Following Partial Ganglionectomy

目的探讨部分背根切除和针刺及内源性c-Fos和c-Jun对体外培养备用背根节(DRG)的作用。方法制备备用根猫模型;采用针刺备用根猫模型、细胞培养以及在部分培养液中加入抗c-Fos/c-Jun抗体的方法,将实验分为针刺组、抗c-Fos抗体组、抗c-Jun抗体组及备用DRG组;并用抗神经元特异性烯醇化酶(NSE)抗体行免疫细胞化学ABC法染色进行神经元鉴定。结果免疫组化染色可见95%以上的细胞为N SE阳性反应的细胞,且为典型的体外培养的DRG神经元。体外培养备用根组、抗c-Fos抗体组、抗c-Jun抗体组神经元突起的平均长度均比针刺组短(P<0.05);抗c-Fos/c-Jun抗体组均比备用根组长(P<0.05)。结论针刺可促进体外培养DRG神经元的突起生长作用;内源性c-Fos、c-Jun可能对体外培养DRG神经元突起生长有促进作用。

Objective To explore the effect of acupuncture, endogenous c-Fos and c-Jun on the regeneration of neuronal dendrite of spared dorsal root ganglion（DRG） in vitro following partial ganglionectomy.Methods Five adult male cats were used in this experiment.Their bilateral L1-L5,L7-S2 DRG were removed,and L6 DRG were spared.Then unilaterally,two sets of acupoints（Zusanli（St.36） and Xuanzhong（G.B.39）;Futu（St.32） and Sanyinjiao（Sp.6） located in the distribution area of spinal nerve L6） were electro-stimulated alternately 30 min everyday by electro-needling.Seven days after operation,bilateral L6 DRGs were taken out and were cultured respectively in vitro.Some cultured mediums of the acupuncture lateral wells were totally replaced by each corresponding antibody-cultured medium including respectively 100 ng/mL anti-c-Fos and anti-c-Jun antibody at the 24^th hour and terminated after 7 days.The length of the neurite was measured by upside-down light microscopy.Then,cultured cells were stained by the immunohistochemistry ABC method.Data were analyzed by One-way ANOVA and q test.Results Immunocytochemical staining revealed that over 95% cells were NSE positive cells which were the typical neuron of DRG in vitro.On the 7^th day,the average neurite length of the spared DRG group,the anti-c-Fos antibody and the anti-c-Jun antibody group were shorter than that of the acupuncture group （P〈0.05）;the average neurite length of the two antibody groups were longer than that of the spared DRG group（P〈0.05）.Conclusion These results indicate that acupuncture,endogenous c-Fos and c-Jun probably promote regeneration of neuronal dendrite of spared DRG in vitro.