双筋龙汤对奥沙利铂所致大鼠后根神经节及神经生长因子表达的影响

Effects of the Shuangjinlong decoction on dorsal root ganglia from Oxaliplatin and nerve growth factor in rats

目的：进一步研究双筋龙汤在奥沙利铂应用期间与后根神经节的形态学改变及DRG 内神经生长因子表达之间的关系，从而为双筋龙汤促进奥沙利铂导致周围神经系统损伤的恢复作用提供依据。方法：将62 只SD 大鼠随机分为空白组、模型组、甲钴胺组、双筋龙汤低、高剂量组。双筋龙汤高、低剂量组分别采用双筋龙汤灌胃，连续10d；甲钴胺组腹腔注射甲钴胺注射液0.135mg/kg，2 次/ 周；模型组、空白组给予双筋龙汤低、高剂量组等体积的生理盐水灌胃，连续10d。第8d 除空白组外均腹腔注射草酸铂20mg/kg，给药草酸铂48h，处死大鼠，取左侧L4 ～ 5 后根神经节。用HE 染色与免疫组织化学的方法检测DRG 形态学改变及NGF 的表达。结果：HE 染色及图像分析，实验组细胞核及核仁发生固缩，可见神经元胞体缩小，胞浆浓缩，与周围细胞分离；部分胞核染色变浅，核膜轮廓模糊甚至消失；核仁碎裂比例增高。免疫组化染色分析，NGF 的表达与双筋龙汤的使用有关，实验组阳性细胞着色较深。结论：双筋龙汤可减轻化疗致大鼠周围神经毒性，其机制可能是通过上调大鼠L4 ～ 5 后根神经节中NGF 的表达来介导。对于奥沙利铂靶器官后根神经节导致的形态学改变作用可减轻其节内感觉神经元的核仁固缩，缓减周围神经系统形态与功能损伤。

Objective: To further investigate the relationship between morphological changes of DRG in the co-administrationof oxaliplatin and the Shuangjinlong decoction and expression of NGF, to provide evidence that the SJLD could recover peripheralnervous system injury from oxaliplatin. Methods: 62 adult female SD rats were divided into the blank group, the model group, themecobalamin group, the Shuangjinlong decoction low-dose group and SJLD high-dose group. The rats in SJLD low dose and highdose groups were treated by irrigation for 10 days. The rats in mecobalamin group were treated with mecobalamin inj ection (0.135mg/kg), twice per week. The model group and the blank group were treated with the same volume saline as high-dose and low-doseSJLD by gavage and lasted for 10 days. Oxaliplatin (20mg/kg) was given to all groups except for the blank group in 8 days. The ratswere sacrificed in 48 hours after Oxaliplatin treatment. The left sides of L4~5 in DRG were harvested. The morphological changes ofDRG and expression of NG were detected with HE staining and immunohistochemistry assay. Results: Through the analysis of HEstaining and image assay, the cell nucleus and kernel in the SJLD groups were pyknosis; meanwhile neurons cell body was reduced.And the cytoplasm were concentrated and separated from the surrounding cells. Some nuclear membranes became lighter and hadhazy outlines, or even disappeared. The ratio of nucleolus fragmentation also were increased. Immunohistochemical staining analysisshows that there is an indeed relationship between the NGF’s expression and SJLD, due to cells stained in the positive group was darker than the others. Conclusion: The Shuangjinlong decoction was effective on chemotherapy-induced neurotoxicity in PNS, andits mechanism may be relevant to up-regulating the expression of NGF in L4~5 of DRG in rats. The morphological effects of thetargets’ dorsal root ganglia within Oxaliplatin can reduce the section of the sensory neurons in nucleolus pyknosis, ease morphologyand the function of the peripheral nervous system injury .