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目的观察运动联合阿仑膦酸钠片(Aln)对去卵巢(OVX)大鼠核因子κB受体活化因子配体(RANKL)/骨保护素(OPG)表达及丝裂原活化蛋白激酶(MAPK)信号通路的影响。方法 90只6月龄雌性SD大鼠随机分为假手术组(sham组)、OVX组、Aln治疗组(OVX+Aln组)、运动治疗组(OVX+EX组)、Aln与运动联合治疗组(OVX+Aln+EX组),每组18只,后4组大鼠切除双侧卵巢。大鼠切除卵巢8周后,各治疗组大鼠分别进行1mg/(kg·d)Aln灌胃和(或)跑台运动干预。干预12周后采用双能X线骨密度仪检测大鼠左、右股骨骨密度(BMD),ELISA法检测大鼠血清RANKL、OPG含量,real-timePCR检测大鼠骨组织RANKL、OPGmRNA表达,蛋白质印迹分析法检测骨组织c-Jun氨基末端激酶(JNK)、p38MAPK(p38)、细胞外调节激酶(ERK)的磷酸化水平及c-Fos表达。结果干预治疗12周后:(1)与sham组相比,OVX组大鼠左、右股骨BMD降低(P<0.05),血清RANKL含量增加、OPG含量降低、RANKL/OPG比值增大(P均<0.05),骨组织RANKLmRNA表达增加、OPGmRNA表达减少(P均<0.05)。(2)与OVX组相比,OVX+Aln组、OVX+EX组及OVX+Aln+EX组大鼠左、右股骨BMD均增加(P均<0.05),血清RANKL含量降低、OPG含量增加、RANKL/OPG比值减小(P均<0.05),骨组织RANKLmRNA表达减少、OPGmRNA表达增加(P均<0.05)。(3)与OVX+Aln组和OVX+EX组相比,OVX+Aln+EX组大鼠左、右股骨BMD,血清RANKL、OPG含量,骨组织RANKL、OPGmRNA表达差异有统计学意义(P均<0.05)。(4)与sham组相比,OVX组大鼠骨组织p-JNK、c-Fos表达增加(P均<0.05);OVX+Aln组、OVX+EX组及OVX+Aln+EX组与OVX组相比,大鼠骨组织p-JNK、c-Fos表达降低(P均<0.05),但3组间表达差异无统计学意义;p-p38、p-ERK在各组间的表达差异无统计学意义。结论运动联合Aln治疗能够提高OVX大鼠左、右股骨骨密度,其作用可能与调节RANKL/OPG平衡、抑制JNK磷酸化及c-Fos表达有关。
Objective To observe the effects of exercise combined with alendronate sodium tablets on the expression of RANKL / OPG and mitogen-activated protein kinase (MAPK) in ovariectomized rats ) Signaling pathways. Methods Ninety 6-month-old female Sprague-Dawley rats were randomly divided into Sham group, OVX group, Aln group (OVX + Aln group), exercise group (OVX + EX group), Aln group and exercise group (OVX + Aln + EX group), each group of 18, the latter 4 groups of rats were removed bilateral ovaries. After ovariectomy in rats for 8 weeks, the rats in each treatment group were given 1 mg / (kg · d) Aln gavage and / or treadmill exercise intervention respectively. Bone mineral density (BMD) of the left and right femurs were measured by dual-energy X-ray absorptiometry after 12 weeks of intervention. The levels of RANKL and OPG were detected by ELISA. RANKL and OPG mRNA expressions were detected by real-time PCR. Protein, The phosphorylation of c-Jun N-terminal kinase (JNK), p38MAPK (p38), extracellular regulated kinase (ERK) and the expression of c-Fos in bone tissue were detected by Western blotting. Results After intervention for 12 weeks: (1) Compared with sham group, BMD of left and right femurs in OVX group decreased (P <0.05), serum RANKL content increased, OPG content decreased, RANKL / OPG ratio increased <0.05). The expression of RANKL mRNA increased and the expression of OPG mRNA decreased (P <0.05). (2) Compared with OVX group, the left and right femur BMD of OVX + Aln group, OVX + EX group and OVX + Aln + EX group were increased (P <0.05), serum RANKL content decreased, OPG content increased, RANKL / OPG ratio decreased (P <0.05). The expression of RANKLmRNA and OPGmRNA in bone tissue decreased (P <0.05). (3) Compared with OVX + Aln group and OVX + EX group, left and right femur BMD, serum RANKL, OPG content, RANKL and OPG mRNA expression in bone in OVX + Aln + EX group were significantly different (P < <0.05). (4) Compared with sham group, the expression of p-JNK and c-Fos in OVX group increased (all P <0.05); OVX + Aln group, OVX + EX group and OVX + Aln + (P <0.05), but the expression of p-JNK and c-Fos was not significantly different among the three groups; there was no statistical difference in the expression of p-p38 and p-ERK Significance of learning. Conclusion Exercise combined with Aln treatment can improve left and right femur bone mineral density in OVX rats, which may be related to the regulation of RANKL / OPG balance and the inhibition of JNK phosphorylation and c-Fos expression.