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目的:探讨恒古骨伤愈合剂(osteoking)对子宫摘除去势骨质疏松症(osteoporosis)树鼩骨细胞增殖的促进作用及通过激活TGF-β/BMP-7信号通路改善骨质疏松症的分子机制。方法:36只雌性实验树鼩随机分为6组,空白对照组不做处理,骨质疏松症模型组摘除子宫(全身骨密度检测下降25%)每天进行生理盐水灌胃,恒古骨伤愈合剂0.05g/kg、0.10g/kg和0.20g/kg灌胃治疗,阿仑膦酸钠0.01g/kg灌胃治疗,10周后处死树鼩,记录体重,收集血清和右下肢骨骼。ELISA法测定各组树鼩血清骨碱性磷酸酶(BALP)、骨钙素(OC)、I型原胶原N-端前肽(PINP),抗酒石酸酸性磷酸酶5b(TRACP-5b)、I型胶原交联C-末端肽(CTX)、雌二醇(E2)、肿瘤坏死因子α(TNFα)、白介素1β(IL-1β)和白介素6(IL-6)的含量;分别采用RT-q PCR和Western blot法检测TGF-β和BMP-7的表达水平。结果:与对照组比较,骨质疏松症组E2含量下降明显,CTX、TRACP-5b、OC、PINP和BALP的含量都显著上升,经恒古骨伤愈合剂治疗后上述现象均有改善效果,且具有剂量依赖性;RT-q PCR与Western Blot结果显示Nrf2和HO-1表达水平经恒古骨伤愈合剂治疗后都有升高趋势,其中当恒古骨伤愈合剂的剂量低于(0.05g/kg)时,以上改善效果均不明显;而当恒古骨伤愈合剂的剂量大于(0.10g/kg)时疗效具有统计学意义,在恒古骨伤愈合剂的剂量达到(0.20g/kg)时治疗组疗效接近于正常组水平。结论:恒古骨伤愈合剂能够有效抑制骨吸收和促进骨形成,并通过激活TGF-β/BMP-7信号通路改善骨质疏松症。
OBJECTIVE: To investigate the effect of osteoking on the proliferation of dendritic cells in ovariectomized osteoporosis and the improvement of osteoporosis by activating TGF-β / BMP-7 signaling pathway Molecular mechanism. Methods: Thirty-six female tree shrews were randomly divided into 6 groups, without treatment in the blank control group. The uterus in the osteoporosis model group (25% reduction in the whole body BMD) was orally administered with normal saline, 0.05 g / kg, 0.10 g / kg and 0.20 g / kg of gavage and gavage of alendronate 0.01 g / kg respectively. After 10 weeks, the tree shrews were sacrificed and the body weight and serum and right lower extremity bone were collected. ELISA was used to detect the serum levels of bone alkaline phosphatase (BALP), osteocalcin (OC), type I procollagen (PINP), tartrate-resistant acid phosphatase 5b (TRACP-5b) (CTX), estradiol (E2), tumor necrosis factor α (TNFα), interleukin 1β (IL-1β) and interleukin 6 (IL-6) PCR and Western blot were used to detect the expression of TGF-β and BMP-7. Results: Compared with the control group, the content of E2 in osteoporosis group decreased significantly and the content of CTX, TRACP-5b, OC, PINP and BALP increased significantly. After the treatment of heal old bone healing agent, The results of RT-q PCR and Western Blot showed that the expression of Nrf2 and HO-1 increased after the treatment of heal-old bone healing agent. When the dose of heel-ancient bone healing agent was lower than 0.05g / kg), the above-mentioned improvement effect was not obvious. When the dose of heel old wound healing agent was more than (0.10g / kg), the curative effect was statistically significant. When the dose of heal old wound healing agent reached 0.20 g / kg), the effect of the treatment group was close to that of the normal group. CONCLUSION: The healing agent of Heng Gu can effectively inhibit bone resorption and promote bone formation, and improve osteoporosis by activating TGF-β / BMP-7 signaling pathway.