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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 observe the effect of Liuweidihuangwan on the signal cascade of JNK and p38MAPK in nucleus pulposus cells in the model of degenerative disc degeneration in rabbits. Methods: Eighty clean-grade New Zealand white rabbits aged 6 months were randomly divided into blank group, sham operation group, model group and Liuweidihuangwan group, 20 in each group. Model group and Liuweidihuangwan group were established animal model of disc degeneration. Liuweidihuangwan group was administrated with Liuwei Dihuang Pill suspension, and the rats in the blank group, sham operation group and model group were given intragastric administration of normal saline. Animals were sacrificed in 4 weeks, 4 weeks, 6 weeks and 8 weeks after the last intragastric administration, L 4/5 and L 5/6 intervertebral discs were removed or paravertebral soft tissues were removed, Maintain the ligament structure of the spine specimens, frozen to be measured. Results: The results of RT-PCR and Western Blot showed that the m RNA content and protein expression of JM, p38MAPK in Liuweidihuangwan group and model group were significantly higher than those in sham operation group and blank control group (P <0.05). There was no significant difference in the mRNA and protein expressions of JNK and P38MAPK in the sham operation group and the blank group (P> 0.05). The m RNA content and protein expression of JNK and P38MAPK in Liuweidihuangwan group were significantly lower than those in the model Group, the difference was statistically significant (P <0.05). Conclusion: Liuweidihuang Pill can inhibit the activation of JNK and p38MAPK pathway, protect the nucleus pulposus cells, stabilize the cells and prevent the degeneration of intervertebral disc. Liuweidihuang Pill can exert the effects of JNK and P38MAPK inhibitors and regulate the expression of JNK and The expression of p38MAPK slowed the apoptosis of nucleus pulposus cells and protected the nucleus pulposus cells.