吡咯喹啉醌通过降低氧化应激和RANKL/OPG比率减少去卵巢大鼠骨量流失研究

Pyrroloquinoline quinone reduces oxidative stress and RANKL/OPG ratio to reduce bone loss in ovariectomized rats

目的观察补充吡咯喹啉醌(PQQ)对去卵巢诱导的骨质疏松大鼠骨量和骨强度的影响。方法30只12周龄(225~260 g)雌性SD大鼠随机分为3组(每组n=10),即OVX组、OVX+PQQ组和Sham组;其中OVX组和OVX+PQQ组进行手术切除双侧卵巢,而Sham组进行假手术;OVX+PQQ组术后食物添加4 mg/kg PQQ。治疗12周,在治疗结束收集血液和股骨,分别进行微型计算机断层扫描(Micro-CT)、生物力学检测、组织切片检测、蛋白印迹检测(WB)以及血清指标检测。结果Micro-CT和HE切片表明,与Sham组相比,OVX组的股骨骨密度和骨微观参数显著降低(P<0.05);OVX组的小梁间距显著增加(P<0.05),而OVX+PQQ上述指标明显优于OVX组(P<0.05)。生物力学检测表明Sham组具有最高的极限载荷、能量和刚度;与OVX组比较,OVX+PQQ组极限载荷、能量和刚度明显增加(P<0.05)。血清学表明,与Sham组大鼠相比,OVX大鼠血清CTX-1、TRACP-5b和MDA水平均显著升高,而T-AOC和SOD活性显著降低;PQQ治疗后显著改善(P<0.05)。WB检测结果表明与Sham组大鼠相比,OVX大鼠RANKL和RANKL/OPG比率均显著升高,而OPG和FOXO1表达显著降低;经过PQQ后得到RANKL和RANKL/OPG比率均显著降低,而OPG和FOXO1表达显著增加(P<0.05)。结论PQQ通过抑制氧化应激和降低RANKL/OPG比率来增加去卵巢大鼠骨量和骨强度。

Objective To observe the effect of(PQQ)supplementation on bone mass and bone strength in ovariectomized rats with osteoporosis.Methods Thirty 12-week-old(225-260 g)female SD rats were randomly divided into three groups:OVX group,OVX+PQQ group and Sham group.Bilateral ovaries were resected in OVX group and OVX+PQQ group,while sham operation was performed in Sham group.4 mg/kgPQQ was added to the food after operation in the OVX+PQQ group.Blood and femurs were collected at the end of treatment for 12 weeks.Microcomputer tomography(Micro-CT),biomechanical examination,tissue section examination,Western Blot(WB)and serum index detection were performed respectively.Results Micro-CT and HE sections showed that the bone mineral density and bone microscopic parameters of HE group were significantly lower than those of Sham group(P<005).The trabecular spacing in OVX group was significantly increased,and the above indexes of OVX+PQQ group were significantly better than those in OVX group(P<005).Biomechanical test showed that the ultimate load,energy and stiffness of Sham group were the highest,and the ultimate load,energy and stiffness of OVX+PQQ group were significantly higher than those of OVX group.Serum CTX-1,TRACP-5b and MDA levels were significantly higher and T-AOC and SOD activities were significantly lower in OVX rats than those in Sham group,but these parameters were improved by supplementation of PQQ.WB test result showed that compared with Sham group rats,the RANKL and RANKL/OPG ratios of OVX rats were significantly increased,while the expressions of OPG and FOXO1 were significantly reduced;after PQQ,RANKL and RANKL/OPG ratios were significantly reduced,while OPG And FOXO1 expression increased significantly(P<0.05).Conclusion PQQ can improve bone mass and bone strength and inhibit bone strength by inhibiting oxidative stress and bone resorption.