灵芝孢子油干预治疗6-羟多巴帕金森病大鼠模型的实验研究

Effect of Ganoderma Lucidum Spores Oil on 6-Hydroxydopamine Rats Model of Parkinson′s Disease

【目的】研究灵芝孢子油对6-羟多巴(6-OHDA)帕金森病(PD)大鼠模型行为学及病理改变的影响,探讨该药对黑质多巴胺能神经元的保护作用。【方法】100只SD大鼠随机分为正常对照组20只、6-OHDA组40只、灵芝孢子油+6-OHDA组40只。通过脑部立体定向法将6-OHDA注射到SD大鼠一侧黑质致密部建立6-OHDA大鼠PD模型,灵芝孢子油+6-OHDA组在造模前3d开始给灵芝孢子油每天500mg/kg,连续10d。6-OHDA组喂食生理盐水做对照。造模后每周给予阿朴吗啡观察大鼠旋转行为的变化,4周后用高效液相色谱法检测大鼠纹状体多巴胺及其代谢物含量,免疫组织化学法检测黑质致密部酪氨酸羟化酶(TH)阳性细胞数量,westernblot法对TH蛋白进行半定量测定。【结果】①灵芝孢子油+6-OHDA组出现旋转行为的大鼠比例为10%,6-OHDA组为45%,两组比较有显著性差异。②纹状体多巴胺及其代谢物含量在两组间存在差异,多巴胺手术侧含量/手术对侧含量在灵芝孢子油+6-OHDA组为(60.12±7.5)%,在6-OHDA组为(38.58±7.26)%。③在黑质致密带区灵芝孢子油+6-OHDA组的TH免疫阳性细胞及TH蛋白表达量均较6-OHDA组显著增多。【结论】灵芝孢子油能明显改善6-OHDA大鼠模型行为学,增加纹状体多巴胺及其代谢物含量并提高黑质多巴胺能神经元的残存率,提示灵芝孢子油可能具有减缓PD病变进程的神经保护作用。

[Objective] To develop neuroprotective treatment that will slow or discontinue the progression of Parkinson′s disease (PD), we explored the ganoderma lucidum spores oil, a rare Chinese crude drug used to cure chronic brain function failure, to evaluate its effect on the pathological condition of 6-hydroxydopamine (6-OHDA) rats model in the substantia nigra pars compacta (SNpc). [Method] SD rats were pretreated with ganoderma lucidum spores oil feeding for 3 days before they were injected with 6-hydroxydopamine into one side substantia nigra, and were continually feeded by ganoderma spores oil for 1 week. The control group rats had not been treated. The animals' cataplasia turning behavior induced by apomorphine were observed every week. Four weeks later, dopamine and its metabolic products in the striatum of both groups were determined by HPLC, and the tyrosine hydroxylase (TH) positive cells were detected by immunohistochemical method,semi-determination expression of TH were detected by Western blot analysis. [Result]① In apormorphine test, rats of the ganoderma spores oil group (10%) presented significant lesser turning than the untreated control group (45%). ② The levels of dopamine in the lesion side striatum were increased in the ganoderma spores oil group. ③ The amount of survival TH positive neurons of the ganoderma spores oil group in SNpc were larger than that of the control group's, either in lesion side or another side. The result of semi-determination indicated that TH expression was significantly increased in the ganoderma spores oil group. [Conclusions] Ganoderma lucidum spores oil can efficiently improve the levels of dopamine in the lesion side striatum, and increase the amount of survival TH neurons as well as TH protein expression in 6-hydroxydopamine rats PD model. It is possible that Ganoderma spores oil may has neuroprotective role to PD.