摘要
目的探讨补骨脂素通过调控无翅型MMTV整合位点家族(wingless type MMTV integration site family,Wnt)信号通路对绝经后骨质疏松症(postmenopausal osteoporosis,PMOP)大鼠的骨保护作用机制。方法将60只SPF级SD雌性大鼠随机分为空白组、模型组、雌二醇片组(0.09mg/kg,每日1次)和补骨脂素低、中、高剂量组(分别为11、22、44mg/kg,每日3次),每组10只。除空白组外,其余组以去卵巢方法构建PMOP大鼠模型。药物干预6周后麻醉取材,行股骨骨密度测定;苏木精-伊红(hematoxylin-eosin,HE)染色观察股骨组织病理形态;逆转录聚合酶链式反应(reverse transcription polymerase chain reaction,RT-PCR)检测大鼠股骨组织Dickkopf相关蛋白1(dickkopf-related protein 1,DKK1)、β-连环蛋白(β-catenin)、Runt相关转录因子2(runt-related transcription factor 2,Runx2)mRNA表达。结果①股骨骨密度方面,与空白组比较,模型组大鼠的双侧股骨骨密度明显降低(P均<0.05);与模型组比较,雌二醇片组、补骨脂素高剂量组大鼠的双侧股骨骨密度及补骨脂素低、中剂量组大鼠的左侧股骨骨密度均明显升高(P均<0.05);与雌二醇片组比较,补骨脂素低剂量组大鼠的双侧股骨骨密度较低(P<0.05)。②股骨组织病理形态方面,与空白组比较,模型组大鼠的股骨骨小梁结构破坏明显且排列紊乱,骨吸收孔增多,骨质疏松特征明显;与模型组比较,雌二醇片组和补骨脂素低、中、高剂量组大鼠的股骨骨小梁结构有所改善,其中,雌二醇片组与补骨脂素高剂量组的骨小梁连续性明显增加,骨陷窝缩小,骨吸收孔减少。③mRNA表达方面,与空白组比较,模型组大鼠股骨组织的DKK1mRNA表达水平明显升高,而β-catenin、Runx2mRNA表达水平明显降低(P均<0.05);与模型组比较,雌二醇片组以及补骨脂素中、高剂量组大鼠股骨组织的DKK1mRNA表达水平明显降低(P<0.05),而β-catenin、Runx2mRNA表达水平明显升高(P均<0.05);与雌二醇片组比较,补骨脂素低剂量组β-catenin、Runx2mRNA表达水平降低(P均<0.05),补骨脂素中、高剂量组大鼠股骨组织的DKK1、β-catenin、Runx2mRNA表达水平差异无统计学意义(P均>0.05)。结论补骨脂素可能通过抑制DKK1表达,进而激活Wnt/β-catenin信号通路,上调Runx2水平,增强骨形成,最终对PMOP大鼠发挥骨保护作用。
Objective To explore the mechanism of the osteoprotective effect of psoralen on postmenopausal osteoporosis(PMOP)rats by regulating wingless type MMTV integration site family(Wnt)signaling pathway.Methods A total of 60SPF grade female SD rats were randomly divided into blank group,model group,estradiol tablet group(0.09mg/kg,once a day)and low-dose psoralen,medium-dose psoralen,high-dose psoralen groups(11,22,44mg/kg,3times a day),with 10rats in each.Except for the blank group,the PMOP rat models were established in the remaining groups by ovariectomy.After 6 weeks of drug intervention,the rats were anesthetized and the samples were collected,the bone mineral density of the femur was measured;the pathological morphology of the femoral tissues was observed by hematoxylin-eosin(HE)staining;the mRNA expressions of dickkopf-related protein 1(DKK1),β-catenin and runt-related transcription factor 2(Runx2)in the femoral tissues of rats were detected by reverse transcription polymerase chain reaction(RT-PCR).Results①In terms of femoral bone mineral density,compared with the blank group,the bilateral femoral bone mineral density of rats in the model group significantly decreased(all P<0.05);compared with the model group,the bilateral femoral bone mineral density of rats in estradiol tablet group and high-dose psoralen group as well as left femoral bone mineral density of rats in low-dose psoralen group and middle-dose psoralen group significantly increased(all P<0.05);compared with the estradiol tablet group,the bilateral femoral bone mineral density of rats in low-dose psoralen group was lower(P<0.05).②In terms of the pathological morphology of femoral tissues,compared with the blank group,the trabecular bone structures of rats in model group was significantly damaged and arranged disorderly,with an increase in bone resorption pores,showing obvious characteristics of osteoporosis;compared with the model group,the trabecular bone structures of rats in estradiol tablet group,low-dose psoralen group,medium-dose psoralen group and high-dose psoralen group were improved,among which,the continuity of trabecular bone in the estradiol tablet group and the high-dose psoralen group significantly increased,the bone lacunaeﾋ=were reduced,and the bone resorption pores decreased.③In terms of mRNA expression,compared with the blank group,the expression level of DKK1mRNA in the femoral tissues of rats in the model group significantly increased,while the expression levels ofβ-catenin and Runx2mRNA significantly decreased(all P<0.05);compared with the model group,the expression levels of DKK1mRNA in the femoral tissues of rats in estradiol tablet group,medium-dose psoralen and highdose psoralen group significantly decreased(P<0.05),while the expression levels ofβ-catenin and Runx2mRNA significantly increased(all P<0.05);compared with the estradiol tablet group,the expression levels ofβ-catenin and Runx2 mRNA decreased in low-dose psoralen group(all P<0.05),there were no statistically significant differences in the expression levels of DKK1,β-catenin and Runx2mRNA in the femoral tissues of rats in medium-dose psoralen and highdose psoralen group(all P>0.05).Conclusion Psoralen may exert an osteoprotective effect on PMOP rats ultimately by inhibiting the expression of DKK1,thereby activating the Wnt/β-catenin signaling pathway,upregulating the level of Runx2and enhancing bone formation.
作者
何田田
唐芳
叶海霞
赵超越
王莎莎
刘欢
HE Tiantian;TANG Fang;YE Haixia;ZHAO Chaoyue;WANG Shasha;LIU Huan(Department of Second Clinical Medical School,Guizhou University of Traditional Chinese Medicine,Guiyang Guizhou 550002,China)
出处
《联勤军事医学》
2025年第3期194-198,共5页
Military Medicine of Joint Logistics
基金
国家自然科学基金项目(82160917)
贵州省高等学校中西医结合防治疾病转化医学重点实验室建设项目[黔教技(2023)017号]
贵州中医药大学国家与省级科技创新人才团队培育项目[贵中医TD合字(2022)004号]
贵州中医药大学大学生创新创业训练计划项目[贵中医大创合字(2021)57号]
