补肾强筋胶囊对肾虚型膝骨关节炎大鼠软骨雌激素受体α和软骨代谢的影响

Effects of Bushen Qiangjin Capsule on cartilage estrogen receptor α and cartilage metabolism of knee osteoarthritis rats with kidney deficiency type

目的:研究补肾强筋胶囊(BSQJC)对肾虚型膝骨关节炎(KOA)大鼠软骨雌激素受体α(ERα)和软骨代谢的影响。方法:选用50只雌性SD大鼠随机分为空白组、模型组、BSQJC组、来曲唑(LTZ)组和LTZ+BSQJC组,每组10只。除空白组外,其余各组采用切除双侧卵巢以及关节腔注射木瓜蛋白酶和半胱氨酸(2∶1)的方法建立肾虚型KOA大鼠模型,造模成功后除空白组和模型组分别给予纯净水灌胃外,其余各组分别给予相应的药物进行灌胃。给药6周后,取大鼠膝关节软骨组织,采用HE染色和番红-固绿染色检测软骨组织。通过ELISA检测大鼠血清Ⅱ型胶原交联羧基端肽(CTX-Ⅱ)和雌二醇(E;)水平,通过RT-qPCR检测大鼠膝关节软骨ACAN、ADAMTS-5、COL2A1、ERα、MMP-13和miR-140-5p基因表达,采用Western Blot法检测大鼠膝关节软骨ACAN、ADAMTS-5、COL2A1、ERα和MMP-13蛋白表达。结果:BSQJC明显减轻肾虚型KOA大鼠软骨组织的病理损伤。并且BSQJC可以显著降低肾虚型KOA大鼠血清CTX-Ⅱ水平,显著提高E;水平(P<0.01),但对LTZ处理后的肾虚型KOA大鼠血清E;水平无显著性影响。另外,无论是否用LTZ进行干预,BSQJC均可显著提高肾虚型KOA大鼠软骨ACAN、COL2A1和ERα基因以及蛋白表达(P<0.05,P<0.01),显著提高miR-140-5p基因表达(P<0.05,P<0.01),并且显著降低ADAMTS-5和MMP-13基因以及蛋白表达(P<0.01)。结论:BSQJC对肾虚型KOA大鼠软骨有一定保护作用,其作用机制不仅与其明显提高体内E;水平有关,而且也与其所含的植物雌激素调控ERα、miR-140-5p表达以及软骨代谢有关。

Objective:To study the effects of Bushen Qiangjin Capsule (BSQJC) on cartilage estrogen receptorα(ERα) and cartilage metabolism of knee osteoarthritis (KOA) rats with kidney deficiency type.Methods:Fifty female SD rats were randomly divided into:control group,model group,BSQJC group,letrozole (LTZ) group and LTZ+BSQJC group,with 10rats in each group.Except the control group,the other groups used the method of bilateral ovariectomy and intra-articular injection of papain and cysteine (2∶1) to establish the KOA rat model with kidney deficiency type.After the model was successfully established,the control group and the model group were given oral administration of pure water respectively,and the other groups were given corresponding drugs by oral administration respectively.After 6 weeks of administration,knee cartilage tissues of rat were taken for HE staining and Safranin O-Fast Green staining.ELISA was used to detect levels of cross linked C-telopeptide of typeⅡcollagen (CTX-Ⅱ) and estradiol (E;) in rat serum,RT-qPCR was used to detect the gene expressions of ACAN,ADAMTS-5,COL2A1,MMP-13,ERαand miR-140-5p,Western Blot was used to detect the protein expressions of ACAN,ADAMTS-5,COL2A1,MMP-13 and ERαin rat knee cartilage.Results:BSQJC notably reduced pathological damage of cartilage tissues in KOA rats with kidney deficiency type.In addition,BSQJC notably reduced CTX-Ⅱlevel and elevated E;level (P<0.01),while it had no significant effect on E;level in serum of LTZ-treated KOA rats with kidney deficiency type.Furthermore,whether treated with LTZ or not,BSQJC significantly increased the gene and protein expressions of ACAN,COL2A1 and ERα(P<0.05,P<0.01),increased the gene expression of miR-140-5p (P<0.05,P<0.01),and reduced the gene and protein expressions of ADAMTS-5 and MMP-13 in KOA rats with kidney deficiency type (P<0.01).Conclusion:BSQJC has a certain protective effect on cartilage of KOA rats with kidney deficiency type,whose mechanism is not only related to the increase of E;level in vivo,but also related to the regulation of ERαand miR-140-5p expressions and cartilage metabolism by phytoestrogens contained in the BSQJC.