目的:观察滋补脾阴方药血清、他克莫司(FK506)及两者联合应用对大鼠背根神经元生长的影响,探讨两种药物对神经细胞的作用机理。方法:取8只新生24hSD大鼠的背根神经节,剥除神经外膜,剪成约1mm3大小的碎块,用0.25%胰蛋白酶消化后进行纯化培养,建立体外新生大鼠背根神经元培养体系。培养96h后,将背根神经元分为空白对照组(A组)、空白血清对照组(B组)、1%滋补脾阴方药血清组(C组)、FK506(1nmol/L)组(D组)和联合用药组(E组)。继续培养48h后,应用四甲基偶氮唑盐(MTT)比色法检测细胞活力,用逆转录聚合酶链式反应法(RT-PCR)检测生长相关蛋白-43信使核糖核酸(GAP-43mRNA)的表达情况。结果:B组的MTT吸光度(OD)值及GAP-43mRNA的相对表达量与A组比较无显著性差异(P>0.05),C组、D组和E组的MTTOD值及GAP-43mRNA的相对表达量均较A组显著增高(P<0.05);C组和D组的MTTOD值及GAP-43mRNA的相对表达量均较B组显著增高(P<0.05),E组与B组比较无显著性差异(P>0.05);D组、E组与C组比较无显著性差异(P>0.05),E组与D组比较有显著性差异(P<0.05)。结论:滋补脾阴方药和FK506对体外培养的大鼠背根神经元具有保护和促再生作用,但两者联合应用时作用明显减弱。 Objective: To observe the effects of nourishing spleen Yin prescription serum (FK506) and their combination on the growth of rat dorsal root neurons and to explore the mechanism of action of the two drugs on nerve cells. Methods: The dorsal root ganglions of 8 neonatal rats with 24hSD were dissected, the epineurium was removed and cut into fragments of about 1mm3. After being digested with 0.25% trypsin, the cultured neurons were purified and cultured to establish dorsal root neurons Training system. The dorsal root neurons were divided into blank control group (group A), blank control group (group B), 1% nourishing spleen Yin prescription serum group (group C), FK506 (1 nmol / L) Group) and combination group (group E). After cultured for 48h, cell viability was measured by MTT colorimetric assay, and growth-associated protein-43 messenger RNA (GAP-43 mRNA) was detected by reverse transcription polymerase chain reaction ) Of the expression. Results: MTT OD value and relative expression level of GAP-43 mRNA in group B were not significantly different from those in group A (P> 0.05). The MTT OD value and relative expression of GAP-43 mRNA in group C, D and E (P <0.05). The MTTOD and the relative expression of GAP-43 mRNA in group C and group D were significantly higher than those in group B (P <0.05), but there was no significant difference between group E and group B (P> 0.05). There was no significant difference between group D, group E and group C (P> 0.05). There was significant difference between group E and group D (P <0.05). Conclusion: Nourishing spleen Yin prescription and FK506 can protect and promote the regeneration of dorsal root neurons cultured in vitro. However, the effect of combination of them is weakened obviously.