补肾活血汤对大鼠腰椎间盘退行性变模型Fas/FasL信号通路的影响

Effects of Bushen Huoxue Decoction on Fas/FasL signaling pathway of lumbar intervertebral disc degeneration rat model

目的 探索补肾活血汤对大鼠腰椎间盘退行性变模型脂肪酸合成酶(fatty acid synthase, Fas)/脂肪酸合成酶配体(fatty acid synthase ligand, FasL)信号通路的影响。方法 50只大鼠随机分成空白组、假手术组、补肾活血汤组、腰痹通组、模型组,每组10只。空白组、假手术组、模型组每日10 mL/kg生理盐水灌胃;补肾活血汤组每日28.98 g/kg补肾活血汤灌胃;腰痹通组每日0.34 g/kg腰痹通灌胃,各组均干预4周。干预结束后,取腰椎间盘组织,番红固绿、Masson染色观察病理学变化,免疫组织化学法计算积分光密度(integral optical density, IOD)值,Western bolt检测Fas、FasL蛋白表达水平,RT-PCR检测Fas、FasL mRNA表达水平。结果 空白组、假手术组细胞形态基本正常,细胞核清晰可见;模型组纤维环碎裂明显,纤维环与髓核间中断,结构不清晰,且染色较深;补肾活血汤组纤维环轻度破裂,髓核细胞及空泡数量略有减少,髓核中基质轻度凝结。与空白组比较,模型组Fas、FasL IOD值均明显升高(P<0.01),Fas、FasL蛋白和mRNA表达水平均明显降低(P<0.01)。假手术组与空白组间Fas、FasL IOD值、蛋白和mRNA表达水平比较,差异均无统计学意义(P>0.05)。与模型组比较,补肾活血汤组、腰痹通组Fas、FasL IOD值均明显降低(P<0.01),Fas、FasL蛋白和m RNA表达水平均明显升高(P<0.01)。与腰痹通组比较,补肾活血汤组Fas、FasL IOD值均明显降低(P<0.01),Fas、FasL蛋白和m RNA表达水平均明显升高(P<0.01)。结论 补肾活血汤能够有效治疗因腰椎退行性变对髓核及纤维环造成的损伤,促进相关蛋白的表达,可能与其调控Fas/FasL信号通路相关。

Objective To study the effects of Bushen Huoxue Decoction(BSHXD) on Fas/FasL signaling pathway of lumbar intervertebral disc degeneration rat model. Methods Fifty rats were randomized into blank group, sham-operated group, BSHXD group, Yaobitong group and model group, with 10 rats in each group. Blank group, sham-operated group and model group were given 10 mL/kg saline by gavage daily;BSHXD group received 28.98 g/kg BSHXD daily by gavage;Yaobitong group was administered with 0.34 g/kg Yaobitong capsule daily by gavage, all groups were intervened for 4 weeks. At the end of the intervention,safranine fast green and Masson staining were used to observe the pathological changes of the lumbar disc tissues, the integral optical density(IOD) values were calculated by immunohistochemistry, Fas and FasL protein expression levels were detected by Western blot and Fas and FasL mRNA expression levels were identified by RT-PCR. Results The cell morphology of specimens in blank and sham-operated groups was normal, and the nuclei were clearly visible;the fibrous rings of model group were obviously fragmented, with interruptions between fibrous rings and nuclei, the structures were not clear, and the staining was quite dark;BSHXD group showed mild rupture of the fibrous rings, a slight decrease in the number of nucleus cells and vacuoles, and mild condensation of the matrix in the nuclei. Compared with blank group, Fas and FasL IOD values were significantly higher(P<0.01)and Fas, FasL protein and mRNA were significantly lower(P<0.01) in model group. The differences in Fas, FasL IOD values, protein and mRNA expression levels between sham-operated group and blank group were not statistically significant(P>0.05). Compared with model group, Fas and FasL IOD values were significantly lower(P<0.01), Fas, Fasl protein and mRNA expression levels were significantly higher(P<0.05). Compared with Yaobitong group, Fas and FasL IOD values were significantly lower(P<0.01), and the expression levels of Fas, FasL proteins and mRNA were significantly higher(P<0.01) in BSHXD group. Conclusion BSHXD can treat the damage to the nucleus pulposus and the fibrous ring caused by degenerative lesions of the lumbar spine, and promote the expression of related proteins. It may be related to the regulation of the Fas/FasL signaling pathway.