基于Akt/PPARγ通路探讨培本固疏方通过促成骨干预绝经后骨质疏松症的机制

Mechanism Exploration of PBGSF in Intervening PMOP by Promoting Bone Formation Based on Akt/PPARγPathway

目的:结合前期研究结果,完善培本固疏方(PBGSF)通过促成骨干预绝经后骨质疏松症(Postmenopausal osteoporosis,PMOP)的具体机制。方法:将36只大鼠随机分为假手术组、模型组、PBGSF低剂量组(低剂量组)、PBGSF中剂量组(中剂量组)、PBGSF高剂量组(高剂量组)和西药组。末次给药后,测定胫骨骨密度(BMD),骨组织弹性位移、极限位移、脆性,骨细胞的矿化水平,Akt/PPARγ的蛋白表达以及血清ALP、ROS的变化。结果:与假手术组比较,模型组大鼠BMD、骨组织弹性及极限位移降低,脆性升高(P<0.05),骨组织未出现钙化结节;Akt的表达水平降低,PPARγ蛋白的表达水平、血清中ALP和ROS含量均显著升高(P<0.01)。与模型组相比,各治疗组BMD均增高(P<0.05,P<0.01),极限位移增高,脆性显著降低(P<0.05,P<0.01);高剂量组、西药组弹性位移、极限位移显著增高(P<0.01),脆性降低(P<0.05)。各治疗组均可见大量钙化结节,低、中、高剂量组骨组织钙沉积现象与给药浓度正相关。低、中剂量组Akt蛋白表达水平均上升(P<0.05),高剂量组、西药组Akt蛋白表达水平显著增高(P<0.01)。低、中、高剂量组和西药组PPARγ蛋白的表达水平降低显著(P<0.01)。各治疗组ROS均明显降低(P<0.01),低、中、高剂量组效应更优,且具有浓度依赖性。结论:培本固疏方能有效改善绝经后骨质疏松,其作用效果在一定范围内具有剂量依赖性。该方通过促进成骨细胞增殖成熟发挥其促成骨作用,并可通过Akt/PPARγ通路以及抗氧化应激作用改善骨质疏松。

Objective:To complete the specific mechanism of Peiben Gushu Formula(PBGSF)in intervening postmenopausal osteoporosis(PMOP)by promoting bone formation on the basis previous research results.Methods:36 rats were randomly divided into the sham operation group,the model group,the PGBSF groups of low-dose,medium-dose and high-dose,as well as the western medication group.After the last administration,the tibia bone mineral density(BMD),bone tissue displacement elasticity and ultimate displacement,bone fragility,bone cell mineralization level,the protein expressions of Akt and PPARγ,as well as the serum changes of ALP and ROS were measured in the groups.Results:Compared to those in the sham operation group,BMD,bone tissue elasticity and ultimate displacement were decreased,and bone fragility was increased in the model group(P < 0.05);no calcified nodules appeared in the model group;the protein expression of Akt was decreased(P < 0.01),the protein expression of PPARγ was increased,and the serum levels of ALP and ROS were also increased significantly in the model group(P < 0.01).Compared to those in the model group,BMD was increased in all treatment groups(P < 0.05,P < 0.01);the ultimate displacement was increased,and the fragility was significantly reduced(P < 0.05,P < 0.01);the elastic displacement and ultimate displacement were significantly increased(P < 0.01),and the fragility was greatly reduced in the PBGSF group of high-dose and in the western medication group(P < 0.05).A large number of calcified nodules were seen in all treatment groups,and the calcium deposition in bone tissue of the PBGSF groups was positively correlated with the administration concentration.The protein expression of Akt was increased in the PBGSF groups of low-dose and medium-dose(P < 0.05),and the protein expression of Akt was significantly increased in the PBGSF group of high-dose and in the western medication group(P <0.01).The protein expression of PPARγ of bone tissue was significantly decreased in the PBGSF groups of medium-dose and high-dose,as well as in the western medication group(P < 0.01).ROS was significantly reduced in each treatment group(P < 0.01),the reductions were more significant in the PBGSF groups of low-dose,medium-dose and high-dose,which showed a concentration-dependent effect.Conclusion:PBGSF can effectively improve PMOP,and its effect is dose-dependent within a certain range.It can promote the proliferation and maturation of osteoblasts to play its role in promoting bone formation.It can improve osteoporosis through Akt/PPAR-γ pathway and anti-oxidative stress.