摘要：

目的: 探讨养帕方对亚急性帕金森病模型小鼠行为学和疼痛症状的治疗效果及其可能的作用机制。方法: 共42只无特定病原体级C57BL/6J小鼠随机分为对照组，模型组，养帕方低剂量组[13.50 g/（kg·d）]、中剂量组[40.50 g/（kg·d）]和高剂量组[121.50 g/（kg·d）]，多巴丝肼组（每组各7只）。采用爬杆实验、悬挂实验和旷场实验评估小鼠行为学改变，热痛反应潜伏期测量仪和电子压力测痛仪测量热刺激伤害性感受阈和机械刺激伤害性感受阈，免疫组化染色检测黑质和杏仁核酪氨酸羟化酶（TH）阳性表达量。结果: 不同处理组小鼠爬杆时间（F=14.625，P=0.000）、悬挂评分（F=24.493，P=0.000）和静止时间（F=24.506，P=0.000），热刺激伤害性感受阈值（F=24.726，P=0.000）和机械刺激伤害性感受阈值（F=21.052，P=0.000），黑质（F=19.663，P=0.000）和杏仁核（F=36.513，P=0.000）TH阳性表达量差异均具有统计学意义。与对照组相比，模型组爬杆时间（P=0.000）和静止时间（P=0.000）延长，悬挂评分（P =0.000）、热刺激伤害性感受阈值（P =0.000）和机械刺激伤害性感受阈值（P =0.000）、黑质（P=0.000）和杏仁核（P=0.000）TH阳性表达量降低。与模型组相比，养帕方低剂量组、中剂量组、高剂量组和多巴丝肼组爬杆时间（P=0.020，0.000，0.000，0.000）和静止时间（P=0.000，0.000，0.000，0.000）缩短，但仍长于对照组（均P < 0.05）；悬挂评分（P=0.000，0.000，0.000，0.000）、热刺激伤害性感受阈值（P=0.008，0.000，0.000，0.000）和机械刺激伤害性感受阈值（P=0.003，0.000，0.000，0.000）、黑质（P=0.031，0.001，0.000，0.000）和杏仁核（P=0.007，0.000，0.000，0.000）TH阳性表达量增加，除多巴丝肼组热刺激伤害性感受阈值恢复至对照组水平（P=0.063）外，其余3组仍低于对照组（均P < 0.05）。与养帕方低剂量组比较，养帕方中剂量组、高剂量组和多巴丝肼组爬杆时间（P=0.009，0.009，0.006）和静止时间（P=0.024，0.018，0.001）亦缩短，但仍长于对照组（均P < 0.05）；悬挂评分（P=0.015，0.011，0.002）、热刺激伤害性感受阈值（P=0.001，0.001，0.000）和机械刺激伤害性感受阈值（P=0.035，0.001，0.001）、黑质（P=0.043，0.023，0.001）和杏仁核（P=0.007，0.005，0.000）TH阳性表达量亦增加，除多巴丝肼组杏仁核TH阳性表达量高于养帕方中剂量组（P=0.009）和高剂量组（P=0.012）外，养帕方中剂量组、高剂量组和多巴丝肼组各项指标差异无统计学意义（均P > 0.05）。结论: 养帕方可以有效改善亚急性帕金森病模型小鼠行为学和疼痛症状，推测是通过保护黑质和杏仁核多巴胺能神经元实现的。

关键词: 帕金森病, 行为和行为机制, 疼痛, 感觉阈, 酪氨酸单氧化酶, 免疫组织化学, 疾病模型, 动物

Abstract:

Objective: Explore the therapeutic effect of Yangpafang on behavior and pain in subacute Parkinson's disease (PD) model mice and its possible mechanism. Methods: Total 42 specific pathogen-free (SPF) C57BL/6J mice were randomly divided into control group, model group, Yangpafang low-dose group [13.50 g/(kg·d)], medium-dose group [40.50 g/(kg·d)] and high-dose group [121.50 g/(kg·d)], levodopa and benserazide group (7 mice in each group). The behavioral changes of mice were evaluated by Pole Climbing Test, Suspension Test and Field Test. The thermal nociceptive threshold and mechanical nociceptive threshold were measured by thermal pain response latency measuring instrument and electronic pressure pain measuring instrument. Immunohistochemical staining was used to detect the positive expression of tyrosine hydroxylase (TH) in substantia nigra and amygdala. Results: There were significant differences in pole climbing time (F =14.625, P=0.000), suspension score (F =24.493, P=0.000), resting time of Field Test (F =24.506, P=0.000), thermal nociceptive threshold (F =24.726, P=0.000) and mechanical nociceptive threshold (F =21.052, P=0.000), TH positive expression in substantia nigra (F =19.663, P=0.000) and amygdala (F =36.513, P=0.000) among different treatment groups. Compared with the control group, the pole climbing time (P=0.000) and resting time (P=0.000) of the model group were prolonged, and the suspension score (P=0.000), thermal nociceptive threshold (P=0.000) and mechanical nociceptive threshold (P=0.000), substantia nigra (P=0.000) and amygdala (P=0.000) TH positive expression decreased. Compared with the model group, the pole climbing time (P=0.020, 0.000, 0.000, 0.000) and resting time (P=0.000, 0.000, 0.000, 0.000) of Yangpafang low-dose group, medium-dose group, high-dose group and levodopa and benserazide group were shortened, but still longer than those of the control group (P < 0.05, for all). The suspension score (P=0.000, 0.000, 0.000, 0.000, 0.000), thermal nociceptive threshold (P=0.008, 0.000, 0.000, 0.000) and mechanical nociceptive threshold (P=0.003, 0.00, 0.000, 0.000), TH positive expression in substantia nigra (P=0.031, 0.001, 0.000, 0.000) and amygdala (P=0.007, 0.000, 0.000, 0.000) increased. The thermal nociceptive threshold in other 3 groups was still lower than that in the control group (P < 0.05, for all), except that the levodopa and benserazide group returned to the level of the control group (P=0.063). Compared with the Yangpafang low-dose group, the pole climbing time (P=0.009, 0.009, 0.006) and resting time (P=0.024, 0.018, 0.001) in the Yangpafang medium-dose group, high-dose group and the levodopa and benserazide group were also shortened, but still longer than those in the control group (P < 0.05, for all). Suspension score (P=0.015, 0.011, 0.002), thermal nociceptive threshold (P=0.001, 0.001, 0.000) and mechanical nociceptive threshold (P=0.035, 0.001, 0.001) also increased, TH positive expression was also increased in the substantia nigra (P=0.043, 0.023, 0.001) and amygdala (P=0.007, 0.005, 0.000). Except that the TH positive expression in the amygdala of the levodopa and benserazide group was higher than that of the Yangpafang medium-dose group (P=0.009) and the high-dose group (P=0.012), there was no significant difference in each index between the latter 3 groups (P > 0.05, for all). Conclusions: Yangpafang can improve the behavior and pain of subacute PD model mice, which is related to the protection of amygdala and substantia nigra dopaminergic neurons.

Key words: Parkinson disease, Behavior and behavior mechanisms, Pain, Sensory thresholds, Tyrosine 3-monooxygenase, Immunohistochemistry, Disease models, animal