基于Nrf2/SIRT3/SOD2途径探讨正清风痛宁抗骨质疏松的作用机制

The mechanism of Zhengqing Fengtongning on anti-osteoporosis based on Nrf2/SIRT3/SOD2 pathway

目的观察正清风痛宁对骨质疏松大鼠的改善作用,并基于Nrf2/SIRT3/SOD2途径探讨其内在的作用机制。方法80只SD大鼠随机分为正常对照组(n=20),骨质疏松组(n=20),正清风痛宁组(n=20),对照药白藜芦醇组(n=20)。除正常对照组外,其余大鼠灌胃醋酸泼尼松[5 mg/(kg·d)]4周诱导骨质疏松症,正清风痛宁组给予正清风痛宁[20 mg/(kg·d)],对照药组给予白藜芦醇[5 mg/(kg·d)],正常对照组及骨质疏松组给予等量0.5%CMC-Na溶液,连续8周。检测各组大鼠血清降钙素(CT)、碱性磷酸酶(ALP)、超氧化物歧化酶2(SOD2)、丙二醇(MDA),血清骨钙素(BGP)、Ⅰ型胶原羧基末端交联肽(β-CTX)水平;骨密度及股骨近端孔隙。HE染色观察股骨病理改变;测定骨组织中Nrf2、SIRT3蛋白的表达情况。结果骨质疏松组大鼠血清CT、ALP、BGP水平及SOD2活性明显降低,β-CTX水平及MDA水平明显升高,骨密度明显降低,股骨近端孔隙率明显升高,HE染色显示骨小梁稀疏、变细、断裂、结构紊乱,骨组织中Nrf2、SIRT3蛋白表达明显降低。与骨质疏松组比较,正清风痛宁组大鼠血清ALP、CT、BGP水平及SOD2活性明显升高,β-CTX及MDA水平明显降低,骨密度明显升高,股骨近端孔隙率明显降低,骨组织结构有明显改善,骨小梁排列整齐且明显增粗,骨组织中Nrf2、SIRT3蛋白表达明显升高。结论正清风痛宁能显著改善醋酸泼尼松诱导的大鼠骨质疏松,其机制可能与Nrf2/SIRT3/SOD2通路作用有关。

Objective To explore the effect and internal mechanism of Zhengqingfengtongning on the osteoporotic rats by Nrf2/SIRT3/SOD2 pathway.Methods 80 male SD rats were randomly divided into normal control group(n=20),osteoporosis group(n=20),Zhengqing Fengtongning group(n=20),control drug group(n=20).Rats were given prednisone acetate(5 mg/kg·d)by 4 weeks induced osteoporosis except for the normal control group,Zhengqing Fengtongning group were given Zhengqing Fengtongning(20 mg/kg·d),control drug group were given resveratrol(5 mg/kg·d),normal control group and osteoporosis group were given 0.5%CMC-NA solution for consecutive 8 weeks.Detected the serum alkaline phosphatase(ALP),calcitonin,serum SOD2,MDA,osteocalcin(BGP),collagen typeⅠcarboxyl terminal crosslinking peptide and bone density,proximal femur pore;The protein expression of Nrf2,SIRT3 of the femoral organization were detected by WB,observe the pathological of the femoral organizatio by HE staining.Results In the model group,serum(ALP),calcitonin,BGP levels and SOD2 activity were significantly reduced,β-CTX levels and MDA levels were significantly increased,bone density was significantly reduced,and proximal femur porosity was significantly increased,HE staining showed sparse bone trabecula,thinning,structure was disorder,the protein expression of Nrf2,SIRT3 were significantly reduced.Compared with the osteoporotic group,in Zhengqing Fengtongning group,the content of ALP,calcitonin,BGP and SOD2 activity were significantly increased,the content of MDA andβ-CTX were significantly reduced,the bone density were significantly increased,the porosity of the proximal femur were significantly reduced,the bone trabeculae were thickened and arranged neatly,and the bone tissue structure were significantly improved.the protein expression of Nrf2 and SIRT3 in the bone tissue were significantly increased.Conclusion Zhengqing Fengtongning can significantly improve osteoporosis in rats induced by prednisone acetate,and its mechanism may be related to the effect of Nrf2/SIRT3/SOD2 pathway.