淫羊藿苷对大鼠椎间盘退变的影响

Effect of icariin on intervertebral disc degeneration in rats

[目的]研究淫羊藿苷(ICA)对大鼠尾椎椎间盘退变(IDD)的影响。[方法]体内实验部分,SD大鼠随机分为假手术组(Sham组)、空白对照组(IDD组)和3个淫羊藿苷处理组(ICAL、ICAM、ICAH)。制备椎间盘退变模型,给予相应处理,并行MIR和细胞因子检测。体内实验部分,自假手术与IDD动物椎间盘髓核分离细胞,体外培养,并给予淫羊藿苷处理(ICAL、ICAM、ICAH),检测细胞和上清的细胞因子等。[结果]体内实验表明,术后2周,与IDD组比较,ICAM组椎间盘T2加权信号强度高;术后6周,IDD组信号消失,ICAM组则无明显改变;ICAM组MRI的Pfirrmann评级均明显低于IDD组(P<0.05)。与Sham组比较,IDD组的IL-1β、IL-6蛋白表达显著增高(P<0.05);与IDD组比较,ICA组的IL-1β、IL-6蛋白表达显著降低(P<0.05)。体外实验显示:IDD组NP细胞增殖率明显降低(P<0.05);而ICA组的NP细胞增殖率明显升高(P<0.05)。与Sham组相比,IDD组NP细胞IL-1β、IL-6表达显著上调(P<0.05);与IDD组比较,ICA各组NP细胞IL-1β、IL-6表达显著下调(P<0.05)。与正常对照组相比,IDD组NP细胞Aggrecan和CollagenⅡmRNA及蛋白表达水平显著降低(P<0.05);与IDD组相比,ICA各组Aggrecan和CollagenⅡmRNA及蛋白表达水平显著升高(P<0.05)。[结论]淫羊藿苷可促进退变髓核细胞增殖、蛋白聚糖及II型胶原蛋白合成,减少IL-1β及IL-6表达,延缓椎间盘退变。

[Objective] To study the effects of icariin(ICA) on the intervertebral disc degeneration(IDD). [Methods] In the study in vivo the SD rats were randomly divided into the sham, IDD, and 3 ICA groups, including low ICA(ICAL), mid ICA(ICAM) and high ICA(ICAH) in dosage. After model creating and corresponding treatments MRI examination and assessments for cytokines were conducted. With respect to the study in vitro, the cells isolated from nucleus pulposus(NP) from the sham and IDD animals were incubate with or without ICA in different dosages, and then the cytokines and other factors in the cells and supernatant were assessed. [Results] As results of the study in vivo, higher signal in T2 weighted images of MRI was noted in ICAM group than the IDD group at 2 weeks after operation. Furthermore, the signal disappeared in the IDD group, whereas that remained unchanged significantly in ICAM group at 6 weeks after operation, associated with significantly lower Pfirrmann grade in ICAM group than the IDD group(P<0.05). The levels of IL-1 and IL-6 detected in the IDD group proved significantly higher than the sham group(P<0.05), while those in the IDD group were also significantly higher than ICA groups(P<0.05). In addition, the NP cell proliferation significantly decreased in the IDD group, whereas significantly increased in the ICA groups in the study in vitro(P<0.05). Regarding to cytokines detected in the cells, the levels of IL-1β and IL-6 significantly increased in the IDD group compared with the sham group(P<0.05), whereas those significantly decreased in the ICA groups in contrast to the IDD group(P<0.05). On the other hand, the expressions of mRNAs of aggrecan and collagen type II in the IDD group significantly declined in the IDD group compared with the sham group(P<0.05), which were significantly reversed by ICA(P<0.05). [Conclusion] The icariin does promote the proliferation of degenerative nucleus pulposus cells, down-regulate the expression of IL-1β and IL-6, and promote the expression of proteoglycan and collagen type II, thereby delaying the degeneration of intervertebral disc.