基于Notch通道右归丸对大鼠退变椎间盘的影响△

Effect of Notch channel-based Yougui Pill on the degenerative intervertebral disc in rats

摘要:

[目的] 探讨右归丸对大鼠退变腰椎间盘结构的潜在干预机制。[方法] 将 30 只雄性 SD 大鼠随机分为 3 组，每组 10 只。正常对照组为健康大鼠，不做特殊处理；退变组采用纤维环穿刺法建立大鼠椎间盘退变模型。药物组在退变组基础上予右归丸灌胃 2 周。ELISA 检测血清炎症因子 IL-10、MIF、TNF-α 水平；Western blot 检测椎间盘组织中 II 型胶原、Notch1 蛋白表达水平。[结果]与正常对照组比较，退变组和药物组的大鼠血清白细胞介素-10（interleukin-10, IL-10）[(22.3±2.8) pg/ml vs (30.9±1.7) pg/ml vs (53.2±7.5) pg/ml, P＜0.001]、巨噬细胞移动抑制因子（macrophage migration inhibitory factor, MIF）[(0.3±0.0) pg/ml vs (1.3±0.3) pg/ml vs (0.6±0.2) pg/ml, P＜0.001]、肿瘤坏死因子 α（tumor necrosis factor-α, TNF-α） [(12.5±3.0) pg/ml vs (52.6±1.5) pg/ ml vs (34.9±3.2) pg/ml, P＜0.001] 含量均显著增加；此外，组织 Notch1 蛋白表达水平显著升高 [(0.1±0.0) vs (0.8±0.1) vs (0.4±0.0), P＜ 0.001]，但是，II 型胶原蛋白表达水平 [(0.9±0.1) vs (0.2±0.0) vs (0.6±0.3), P＜0.001] 显著降低。与退变组相比，药物组的大鼠血清炎症因子 IL-10 含量显著增加 （P＜0.05），但 MIF、TNF-α 含量显著减少 （P＜0.05），II 型胶原蛋白表达水平升高 （P＜0.05）。[结论]右归丸能够抑制椎间盘退变大鼠的炎症反应，减少细胞外基质降解，从而缓解大鼠椎间盘退变，其机制可能与调控 Notch 信号通路相关。

Abstract:

[Objective] To investigate the potential intervention mechanism of Yougui Pills on the degenerative lumbar disc structure in rats. [Methods] Thirty male SD rats were randomly divided into 3 groups with 10 rats in each group. The normal control group was healthy rats without special treatment, while, the model group had intervertebral disc degeneration established by annulus fibrosus punctured. In addition, the drug group was given Yougui pill for 2 weeks on the basis of the model group. Serum levels of inflammatory cytokines, including IL-10, MIF and TNF-α were detected by ELISA, while the expression levels of type II collagen and Notch1 protein in intervertebral disc tissues were detected by western blot. [Results] Compared with normal control group, the serum interleukin-10 (IL-10) [(22.3±2.8) pg/ml vs (30.9±1.7) pg/ml vs (53.2±7.5) pg/ml, P＜0.001], macrophage migration inhibitory factor (MIF) [(0.3±0.0) pg/ml vs (1.3±0.3) pg/ml vs (0.6± 0.2) pg/ml, P＜0.001] and tumor necrosis factor-α (TNF-α) [(12.5±3.0) pg/ml vs (52.6±1.5) pg/ml vs (34.9±3.2) pg/ml, P＜0.001], additionally, the tissue expression of Notch1 protein significantly increased [(0.1±0.0) vs (0.8±0.1) vs (0.4±0.0), P＜0.001], while the type II collagen protein expression decreased significantly [(0.9±0.1) vs (0.2±0.0) vs (0.6±0.3), P＜0.001] in the model group and drug group. However, the drug group proved significantly higher serum IL-10 (P＜0.05), whereas significantly lower MIF and TNF-α (P＜0.05), as well as significantly higher tissue expression of type II collagen than the model group (P＜0.05). [Conclusion] TheYougui Pill can inhibit the inflammatory response and reduce the degradation of extracellular matrix in rats with intervertebral disc degeneration, thus alleviating the intervertebral disc degeneration in rats. The mechanism may be related to the regulation of Notch signaling pathway.