强骨合剂通过Wnt3a/β-catenin/VEGF信号通路促进血管生成改善围绝经期大鼠骨质疏松的研究

Qianggu Decoction Promotes Angiogenesis through Wnt3a/β-catenin/VEGF Pathway to Improve Osteoporosis in Perimenopausal Rats

    摘要
  • 摘要:
      目的  探讨强骨合剂对围绝经期骨质疏松SD大鼠的疗效及其可能的作用机制。
      方法  采用HPLC建立强骨合剂标准指纹图谱。将50只SD大鼠随机分为假手术组、模型组、雌二醇组、强骨合剂高剂量(12 g · kg-1)组、低剂量(6 g · kg-1)组, 每组10只。双侧卵巢切除法制备围绝经期大鼠骨质疏松模型, 分别灌胃给予生理盐水，雌二醇，高、低剂量强骨合剂, 连续给药30 d。记录大鼠体质量和子宫质量, 检测大鼠骨力学相关指标, 股骨HE染色观察成骨细胞量及骨小梁面积。qPCR检测Wnt3a、β-catenin、VEGF、OPG mRNA表达水平; 免疫荧光检测VEGFR-2、CD31, 评价血管生成情况; Western blot检测股骨组织Wnt3a、p-GSK3β、VEGF、β-catenin、Lamin蛋白表达情况。
      结果  10批次提取的强骨合剂指纹图谱相似度均大于0.9, 表明10批样品质量稳定。与模型组比较, 雌二醇组，强骨合剂高、低剂量组可显著改善大鼠子宫指数、骨密度、骨力学性能指标(P < 0.05, P < 0.01), 明显改善骨微结构, 提高股骨组织中Wnt3a、β-catenin、VEGF、OPG mRNA水平(P < 0.05, P < 0.01), 以及VEGF、CD31、Wnt3a、p-GSK3β、β-catenin、Lamin蛋白表达水平(P < 0.05, P < 0.01)。
      结论  强骨合剂可能通过Wnt3a/β-catenin信号通路调节VEGF表达, 促进血管生成, 从而提高大鼠骨密度, 改善骨组织形态, 进而发挥抗骨质疏松的作用。

     

    Abstract:
      OBJECTIVE  To investigate the efficacy and possible mechanism of action of the Chinese medicinal preparation Qianggu decoction on perimenopausal osteoporotic SD rats.
      METHOD  HPLC was used to establish the standard fingerprint profile of Qianggu decoction. Fifty SD rats were randomly divided into sham-operated group, model group, estradiol valerate group, and high (12 g · kg-1) and low (6 g · kg-1) dose groups of Qianggu decoction, with 10 rats in each group. The perimenopausal osteoporosis rat model was prepared by bilateral ovariectomy method, and saline, estradiol or high and low doses of Qianggu decoction were administered by gavage for 30 days. The weight and uterine weight of the rats were recorded, and the bone mechanics related indexes were measured, and the number of osteoblasts and the area of bone trabeculae in the femur were observed by HE staining. Wnt3a, β-catenin, VEGF and OPG gene expression levels were detected by PCR; VEGFR-2 and CD31 were detected by immunofluorescence, and angiogenesis was evaluated. The protein expression of Wnt3a, p-GSK3β, VEGF, β-catenin, Lamin were detected by Western blot.
      RESULTS  The fingerprint similarity of the 10 batches of Qianggu decoction was greater than 0.9, indicating that the quality of the 10 batches of samples was stable. Compared with the model group, estradiol valerate-treated group and high and low dose Qianggu decoction-treated groups could significantly improve the uterine index, bone density and bone mechanical properties of rats(P < 0.05, P < 0.01), significantly improve bone microstructure, and increase the levels of Wnt3a, β-catenin, VEGF, OPG mRNA in femoral tissue(P < 0.05, P < 0.01), as well as VEGF, CD31, Wnt3a, p-GSK3β, β-catenin, Lamin protein expressions(P < 0.05, P < 0.01).
      CONCLUSION  Qianggu decoction may regulate VEGF expression and promote angiogenesis through Wnt3a/β-catenin signaling pathway, thus increasing bone density and improving bone tissue morphology in rats, and then exerted anti-osteoporosis effects.

     

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