骨髓基质细胞对离体Parkinson鼠脑片单胺类神经递质的影响

INFLUENCES ON THE MONOAMINE NEUROTRANSMITTERS BY CO-CULTURING BONE MARROW STROMAL CELLS ONTO BRAIN SLICE OF PARKINSON’ RAT

为探讨骨髓基质细胞对离体Parkinson鼠脑片单胺类神经递质的影响,本研究通过建立新生大鼠离体“Parkinson鼠病”模型,取成年大鼠骨髓,培养、分离、纯化骨髓基质细胞,将骨髓基质细胞(MSCs)与离体脑片联合培养,应用高效液相法观察了脑片和培养液中多巴胺及其代谢产物3,4-二羟苯乙酸(DOPAC)和高香草酸(HVA)含量的变化。结果显示,经MPTP代谢产物mpp+损伤的脑片多巴胺含量与正常组、联合培养组相比明显减少(P<0.01),而mpp+组培养液内DOPAC和HVA含量也减少(P<0.05)。但联合培养组与正常组相比DA,DOPAC和HVA的变化无显著差异。以上结果提示,联合培养的骨髓基质细胞对mpp+毒性损伤的多巴胺能神经元具有保护作用,该模型也为在脑片上测量单胺类神经递质提供了可靠的方法。

To study the effect of monoamine neurotransmitters on brain slice of Parkinson＇s disease （PD） rats in vitro and co-culture with bone marrow stromal cells （ MSCs）, the model of PD in vitro from newborn rat was established. MSCs obtained from adult rats bone marrow were cultured, isolated and purified. MSCs were co-cultured with brain slice from PD rat. The content change of dopamine hydrochloride （DA）, 3,4-dihydroxyphenylacetic acid （DOPAC）and homovanillic acid（HYA） in the tissue and culture medium were detected under high performance liquid chromatography. The results showed that the content of dopamine in the brain slice of MPTP metabolite 1 -methyl- 4-phenylpyridinnium（ mpp^＋） toxicity was obviously decreased comparing with normal group and co-cultured group （ P 〈0.01 ） , while the content of DOPAC and HVA in the culture medium of the mpp^＋ group was also decreased （P 〈 0.05 ）. However, no siginificant changes of DA, DOPAC and HVA levels were detected in co-culture group compared with the normal group. These results suggest that co-culture with MSCs may protect dopaminergic neurons against mpp^＋ toxicity. The present model has been successfully applied a reliable method to measure monoamine neurotransmitters in brain slice.