益气活血方对大鼠腰椎间盘突出的影响及机制

Effects and mechanism of Yiqi huoxue decoction on lumbar disc herniation in rats

目的探讨益气活血方(YQHX)对大鼠腰椎间盘突出的影响及机制。方法大鼠随机分为假手术组、模型组、核因子κB(NF-κB)抑制剂组(QNZ组,1 mg/kg)、YQHX组(9.1 g/kg)及联合组(YQHX+QNZ组,剂量同各单药组),每组10只。除假手术组外,其余各组大鼠建立腰椎间盘突出症模型,各给药组大鼠腹腔注射QNZ和/或灌胃YQHX,每天给药1次,连续8周。评估各组大鼠椎间盘突出严重程度,观察椎间盘组织形态学变化、髓核细胞自噬变化,检测血清中肿瘤坏死因子α(TNF-α)水平、椎间盘组织中B细胞淋巴瘤2/腺病毒E1B相互作用蛋白3(BNIP3)、苄氯素1(Beclin-1)阳性细胞百分比,核因子κB(NF-κB)p65磷酸化水平,肿瘤坏死因子受体相关因子2(TRAF-2)、TRAF-3、BNIP3、微管相关蛋白轻链3B(LC3B)蛋白表达水平,以及NF-κB p65、LC3B、p62、BNIP3及Beclin-1 mRNA表达水平。结果与模型组比较,YQHX组大鼠椎间盘Pfirrmann分级评分显著降低,病理损伤减轻;髓核细胞自噬小体数量增加;血清中TNF-α水平和椎间盘组织中p62 mRNA表达水平均显著降低;椎间盘组织中BNIP3、Beclin-1阳性细胞百分比,NF-κB p65磷酸化水平,TRAF-2、TRAF-3、BNIP3、LC3B蛋白表达水平,NF-κB p65、LC3B、BNIP3、Beclin-1 mRNA表达水平均显著升高(P<0.05)。加入QNZ可部分逆转YQHX组上述指标的变化。结论YQHX可能通过激活NF-κB信号通路促进大鼠椎间盘自噬水平升高,减轻炎症反应,延缓腰椎间盘突出症进展。

OBJECTIVE To investigate the effects and mechanism of Yiqi huoxue decoction(YQHX)on lumbar disc herniation in rats.METHODS Rats were randomly divided into sham operation group,model group,NF-κB inhibitor group(QNZ group,1 mg/kg),YQHX group(9.1 g/kg)and combination group(YQHX+QNZ group,the same dose as each single drug group),with 10 rats in each group.Except for sham operation group,lumbar disc herniation model of rats was induced in other groups;administration groups were given QNZ intraperitoneally or/and YQHX intragastrically,once a day,for 8 consecutive weeks.The severity of intervertebral disc herniation was evaluated in each group;the pathological changes of intervertebral discs and the changes of autophagy of nucleus pulposus cells were all observed;the level of tumor necrosis factor-α(TNF-α)in serum,and the ratios of Bcl-2/adenovirus E1B interacting protein 3(BNIP3)and Beclin-1 positive cells in intervertebral disc tissues were detected;the phosphorylation of nuclear factor-κB(NF-κB)p65,the expressions of tumor necrosis factor receptorassociated factor-2(TRAF-2),TRAF-3,BNIP3 and LC3B protein,and mRNA expressions of NF-κB p65,LC3B,p62,BNIP3 and Beclin-1 were determined.RESULTS Compared with model group,Pfirrmann grading score decreased significantly,the pathological injury of intervertebral disc tissue was relieved in YQHX group;the number of autophagosomes in nucleus pulposus cells increased;serum level of TNF-αand mRNA expression of p62 in intervertebral disc tissue decreased significantly;the ratios of BNIP3 and Beclin-1 positive cells,the phosphorylation of NF-κB p65,the expressions of TRAF-2,TRAF-3,BNIP3 and LC3B protein as well as the mRNA expressions of NF-κB p65,LC3B,BNIP3 and Beclin-1 decreased significantly in intervertebral disc tissues(P<0.05).The changes of above indexes in YQHX group were reversed partly in YQHX+QNZ group.CONCLUSIONS YQHX promotes the elevation of autophagy level of intervertebral discs,slows down the inflammatory response and the progression of lumbar disc herniation by activating the NF-κB signaling pathway.