杜仲腰痛丸调控p53对老年骨质疏松症患者骨密度、骨代谢及骨形成影响的作用机制

The mechanism of Eucommia Lumbago Pill in regulating the effect of p53 on bone mineral density,bone metabolism and bone formation in elderly patients with osteoporosis

目的探讨杜仲腰痛丸调控p53对老年骨质疏松症(osteoporosis,OP)患者骨密度(bone mineral density,BMD)、骨代谢及骨形成影响的作用机制。方法选取自2018年3月至2019年8月期间,在甘肃省中医院门诊就诊的老年骨质疏松症患者30例,口服碳酸钙D3咀嚼片(钙尔奇D)+骨化三醇胶丸(罗钙全)+杜仲腰痛丸(eucommia lumbago pill,ELP)治疗6个月;并选取同期于本院体检健康者30名为对照组。分别用0.2、0.4、0.8 mg/mL ELP培养液处理p53基因过表达的成骨细胞MC3T3-E1。通过RT-PCR及Western blot检测p53、ALP、OCN及Runx 2 mRNA与蛋白的表达水平;进行骨密度及骨代谢生化指标检测并分析与p53表达的相关性。结果与对照组相比,OP患者的P53 mRNA与蛋白的表达量及BGP、TRAP、DPD/CR显著增加,各部位BMD显著降低;ELP治疗6个月后,p53的mRNA与蛋白的水平及BGP、TRAP、DPD/CR均明显降低,而L2~4BMD显著升高。相关性分析表明,p53的表达水平与L2~4BMD含量呈显著负相关,而与BGP、BALP及DPD/CR含量呈显著正相关。与对照组相比,不同浓度的ELP明显降低MC3T3-E1中p53的表达水平;同时0.4 mg/mL的ELP可以逆转p53过表达的影响及对ALP、OCN和Runx 2的表达。结论杜仲腰痛丸可能通过抑制p53的表达影响老年骨质疏松症患者的骨密度、骨代谢,促进骨形成。

Objective To explore the mechanism of Eucommia Lumbago Pill’s regulation of p53 on bone mineral density,bone metabolism and bone formation in elderly patients with osteoporosis(OP).Methods From March 2018 to August 2019,30 elderly patients with osteoporosis(OP)who were treated in the outpatient clinic of Gansu Provincial Hospital of Traditional Chinese Medicine were treated with calcium Erqi,Luoquanquan+oral Eucommia Lumbago Pills(ELP)treated for 6 months,and selected 30 healthy people who were examined in our hospital during the same period as the control group.Osteoblasts MC3T3-E1 with p53 gene overexpression were treated with 0.2,0.4,and 0.8 mg/mL ELP medium.RT-PCR and Western blot were used to detect the expression levels of p53,ALP,OCN and Runx2 mRNA and protein;bone mineral density and bone metabolic biochemical indicators were detected and the correlation with p53 expression was analyzed.Results Compared with the control group,the expression levels of P53 mRNA and protein and BGP,TRAP,DPD/CR in serum samples of OP patients were significantly increased,while BMD in each was significantly reduced.ELP treatment for 6 months After that,the levels of p53 mRNA and protein and BGP,TRAP,DPD/CR were significantly reduced,while L2-4BMD was significantly increased.Correlation analysis showed that the expression level of p53 was significantly negatively correlated with the content of L2-4BMD,but significantly positively correlated with the content of BGP,BALP and DPD/CR.Compared with the Control group,different concentrations of ELP significantly reduced the expression level of p53 in MC3T3-E1.Meanwhile,0.4 mg/mL ELP reversed the effect of p53 overexpression and the expression of ALP,OCN and Runx2.Conclusion Eucommia Lumbago Pill may affect the bone mineral density,bone metabolism balance and promote bone formation in elderly osteoporosis patients by inhibiting the expression of p53.