基于Wnt/β-cat信号通路探讨蒙药润骨灵治疗绝经后骨质疏松症小鼠的疗效及作用机制

To investigate the effect and mechanism of Mongolian medicine Runguling on postmenopausal osteoporosis mice based on Wnt/β-cat signaling pathway

目的:研究蒙药润骨灵对小鼠去卵巢绝经后骨质疏松症(Postmenopausal osteoporosis,PMOP)的疗效及作用机制。方法:选取SPF级6月龄未曾交配雌性C57BL/6小鼠70只,去卵巢法(Ovariectomy,OVX)建立小鼠PMOP模型,每组随机10只:正常组(N),假手术组(S),模型组(M),润骨灵高剂量组(RGL-H),润骨灵中剂量组(RGL-M),润骨灵低剂量组(RGL-L),阿仑膦酸钠片组(AS)。N组和S组之外其余组进行双侧卵巢切除术,灌胃治疗12周后,ELISA法检测血清PINP、BALP、β-CTX、TRACP含量,骨组织和肾组织Wnt3a蛋白含量,RT-PCR法对各组小鼠肾及股骨组织中Wnt3a mRNA及β-catenin的表达进行检测。结果:一般情况,N组小鼠活动活跃、机灵、毛发光顺正常;手术各组小鼠精神较为萎靡,术后体重与N组相比明显下降(P<0.05)。治疗后子宫重量,与N组,S组对比模型组及其他给药组有显著差异(P<0.05)。各组小鼠骨密度BMD值、血清PINP、BALP、β-CTX、TRACP、骨组织和肾组织Wnt3a蛋白含量、Wnt3a mRNA及β-cateni蛋白检测结果:与N组和S组比较M组和各给药组均有下降(P<0.05);与M组对比各给药组均有显著差异(P<0.05);与RGL-M组对比RGL-H组,RGL-L组有明显差异(P<0.05);与AS组对比M组,RGL-M组有显著差异(P<0.05);结论:蒙药润骨灵可以改善骨细胞代谢情况,起到预防和治疗绝经后骨质疏松症的作用。

Objective To investigate the effect and mechanism of Mongolian medicine Runguling for Postmenopausal osteoporosis(PMOP)after ovariectomized mice.Methods:Seventy 6-month-old female C57BL/6 mice with SPF grade were selected to study PMOP model with Ovariectomy(OVX),and 10 mice were selected in each group:normal group,sham group,model group,Runguling high-dose group,Runguling medium-dose group,Runguling low-dose group,alendronate sodium tablet group.The other groups except the normal group and the sham group underwent bilateral ovariectomy.After 12 weeks of intragastatic treatment,serum PINP,BALP,β-CTX,TRACP contents,and Wnt3a protein contents in bone and kidney tissues were detected by ELISA.The expression of Wnt3a mRNA andβ-catenin in kidney and femur tissues of mice in each group were detected by RT-PCR.Results:In general,the mice in the normal group were active,clever,smooth and normal.The spirits of the mice in the operation group were lower than those in the normal group(P<0.05).Uterine weight after treatment was significantly different from that of normal group,sham group and model group as well as other administration groups(P<0.05).Bone mineral density BMD,serum PINP,BALP,β-CTX,TRACP,Wnt3a protein content,Wnt3a mRNA andβ-cateni protein content in bone and kidney of mice in each group were decreased compared with normal and sham operation groups(P<0.05).Compared with model group,all administration groups had significant differences(P<0.05).Compared with Runguling medium dose group,Runguling high dose group and Runguling low dose group had significant differences(P<0.05);Compared with alendronate sodium group,Runguling medium dose group had significant difference(P<0.05);Conclusion:Mongolian medicine Runguling can improve bone cell metabolism and play a role in the prevention and treatment of postmenopausal osteoporosis.