【摘要】 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。更多还原

【Abstract】 Objective:To observe the effects of Zi-Bu-Pi-Yin recipe,tacrolimus(FK506) and their combined use on the growth of dorsal root ganglia neuron(DRGn) in newborn rats and discuss their inner mechanisms.Method:The dorsal root ganglia(DRG) in eight newborn SD rats(within 24 hours) were harvested and the epineurium of the DRG was isolated.After dissected into pieces of about 1mm3 size,the DRG were digested by 0.25% trypsin.Then DRGn were purified and cultured.After cultured for 96 hours,the DRGn were assigned into 5 groups as follows,blank-control group(group A),blank-serum control group(group B),Zi-Bu-Pi-Yin recipe group(1%,group C),FK506 group(1nmol/L,group D) and combined use group(group E).After culturing the DRGn for additional 48 hours,the vitality of the DRGn was examined by MTT method and the expression of GAP-43′s mRNA was tested by RT-PCR respectively.Result:TThere was no statistical difference as for the OD value of MTT and the expression of GAP-43′s mRNA between group B and group A(P>0.05),the OD value of MTT and the expression of GAP-43′s mRNA in group C,D and E were significantly higher than those in group A(P<0.05);the OD value of MTT and the expression of GAP-43′s mRNA in group C and D were significantly higher than those in group B(P<0.05).There was no statistical difference between group E and group B(P>0.05);no significant difference was noted between group D,E and group C(P>0.05) and significant difference was observed between group E and D either(P<0.05).Conclusion:Both Zi-Bu-Pi-Yin recipe and FK506 have effects of neuroprotection and neuroregeneration on the DRGn in newborn rats in vitro,which weakened when combined use.更多还原