六味地黄丸对兔椎间盘退变模型髓核细胞p38MAPK信号级联JNK通路的影响

Effect of Liuwei Dihuang Wan(六味地黄丸) on Expression of JNK and P38 Signaling Pathway of the Intervertebral Disc Injury Rabbit

目的:观察六味地黄丸对兔椎间盘退变模型髓核细胞JNK与p38MAPK信号级联的影响。方法:选取6月龄清洁级新西兰兔80只,随机分为空白组、假手术组、模型组、六味地黄丸组,每组20只。模型组、六味地黄丸组建立椎间盘退变动物模型,六味地黄丸组予六味地黄丸混悬液灌胃,空白组、假手术组、模型组灌以等量生理盐水灌胃。于最后1次灌胃后4 h分2周、4周、6周、8周分批次处死动物,留取L_(4/5),L_(5/6)椎间盘标本或去除椎旁软组织、保留韧带结构的脊柱标本,冻存待测。结果:RT-PCR和Western Blot检测结果表明,六味地黄丸组与模型组兔髓核细胞JM、p38MAPK的m RNA含量和蛋白表达明显高于假手术组与空白组,差异有统计学意(P<0.05);假手术组与空白组JNK、P38MAPK的mRNA含量和蛋白表达比较,差异无统计学意义(P>0.05);六味地黄丸组JNK、P38MAPK的m RNA含量和蛋白表达明显低于模型组,差异有统计学意义(P<0.05)。结论:六味地黄丸能抑制JNK与p38MAPK通路的全面激活,保护髓核细胞,使细胞稳定,对椎间盘退变有明显的防治作用;六味地黄丸可能发挥类JNK与P38MAPK抑制剂作用,调节JNK与p38MAPK的表达,减缓了髓核细胞的凋亡,保护了髓核细胞。

Objective： To explore the efficacy of Liuwei Dihuang Wan（六味地黄丸,LWDHW） on expression of JNK and P38 signaling pathway of the intervertebral disc injury rabbit.Methods： The 80 New Zealand white female rabbits（6 month old） were randomly divided into normal group,sham operation group,model group,LWDHW group,20 rabbits in each group.The animal models of intervertebral disc degeneration were established.LWDHW group was given for LWDHW suspension gavage,normal group,sham operation group and model group was given for the same amount NS,once a day.In the last 4 hours after intragastric administration in 2 weeks,4 weeks,6 weeks,8 weeks the animals were killed in batches,The L_（4/5） and L_（5/6） nucleus pulposus and annulus fibrosus were collected frozen for test.Results： Reverse transcription-PCR and Western Blot showed that with the culture time prolonging,the expressions of JNK collagen and genes in nucleus pulposus and p38 collage and genes in the annulus fibrosus in model group,LWDHW group were significantly higher than normal group and sham operation group,the difference was statistics significant（P〈0.05）.The expressions of JNK collagen and genes in nucleus pulposus and p38 collage and genes in the annulus fibrosus in sham operation group are not obvious different with normal group,there is no statistics significance（P〉0.05）.LWDHW group of the indicates were lower than model group,the difference was statistics significant（P〈0.05）.Conclusion： Liuwei Dihuang Wan can protect the intervertebral discs through stabilizing the expressions of JNK collagen and genes in nucleus pulposus and p38 collage and genes.