中国骨质疏松杂志

滋阴补肾方对糖尿病合并去卵巢骨质疏松大鼠的作用及对Wnt/β-catenin信号的影响

Effect of nourishing Yin and tonifying kidney recipe on the treatment of diabetes mellitus combined with osteoporosis after ovariectomy and the Wnt/beta-catenin signal pathway in rats

DOI：10.3969/j.issn.1006.7108.2020.03.024

中文关键词: Wnt/β-catenin 自噬 NF-κB

英文关键词:Wnt/β-catenin autophagy NF-κB

基金项目:湖北省卫计委科研项目（项目名称：滋阴补肾方对糖尿病合并骨质疏松大鼠的治疗及信号通路的影响）

作者	单位
孙勤国1 徐鸿婕1 罗蒙1 江波1* 吕琨1 杨倩1 黄天慧2 张贤梅1	1．武汉大学同仁医院（武汉市第三医院）中医科，湖北武汉 430060 2．湖北中医药大学第一临床学院，湖北武汉 430060

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中文摘要:

目的探索滋阴补肾方对糖尿病合并去卵巢骨质疏松大鼠治疗及信号通路的影响，为骨质疏松的治疗奠定基础。方法 50只SPF级雌性SD大鼠被分为正常对照组（10只），模型组（8只），骨化三醇组（8只），滋阴补肾方低剂量组（8只），滋阴补肾方中剂量组（8只），滋阴补肾方高剂量组（8只）。投喂高糖高脂饲料并注射链脲佐菌素建立糖尿病合并去卵巢骨质疏松大鼠模型。造模成功后，滋阴补肾方低剂量组，滋阴补肾方中剂量组，滋阴补肾方高剂量组分别灌胃滋阴补肾方5、10、20 g/(kg?d)，骨化三醇组大鼠每天灌胃0.1 μg/(kg?d)骨化三醇，灌胃给药8周，每日1次。给药结束后，以HE染色的方法观察胫骨结构变化；以Western blot的方法检测p-NF-κB、NF-κB、IkB、MyD88的蛋白表达水平；以RT-PCR的方法检测Wnt-3α、LRP-5、β-catenin的表达。结果和模型组比较，滋阴补肾方高剂量组胫骨骨小梁排列较密集，骨小梁之间连接增多；中剂量组和高剂量组的p-NF-κB、NF-κB、IkB、MyD88表达水平均降低，TNF-α、IL-6、IL-1β、IL-8、IL-10水平显著降低，Wnt-3α、LRP-5、β-catenin表达水平均显著升高（P<0.05）。结论滋阴补肾方治疗糖尿病合并去卵巢骨质疏松大鼠可显著增加其成骨细胞的数量，增加骨量，可能上调Wnt/β-catenin信号通路来实现的。

英文摘要:

Objective To explore the effect of nourishing Yin and tonifying kidney recipe on the treatment of diabetes mellitus combined with osteoporosis after ovariectomy and the signal pathway in rats, and to lay a foundation for the treatment of osteoporosis. Methods Fifty SPF-grade female SD rats were divided into control group (n=10), model group (n=8), calcitariol group (n=8), low-dose nourishing Yin and tonifying kidney recipe group (n=8), middle-dose nourishing Yin and tonifying kidney recipe group (n=8), and high-dose nourishing Yin and tonifying kidney recipe group (n=8). The model group was fed with high-sugar and high-fat diet and injected with streptozotocin to establish a diabetic rat model with ovariectomized osteoporosis. After successful modeling, rats in the low-dose nourishing Yin and tonifying kidney recipe group, the middle-dose nourishing Yin and tonifying kidney recipe group, and the high-dose nourishing Yin and tonifying kidney recipe group received 5g/(kg?d), 10g/(kg?d), and 20g/(kg?d) of nourishing Yin and tonifying kidney recipe intragastrically, respectively. Rats in calcitariol group recieved 0.1 g/kg/d of calcitariol per day. The rats were gavaged once per day for 8 weeks. After the treatment, the structure change of the tibia was observed with HE staining. The protein expressions of p-NF-κB, NF-κB, IkB, and MyD88 were detected with Western blotting. The expressions of Wnt-3α, LRP-5, and β-catenin were detected using RT-PCR. Results Compared with those in the model group, the tibial trabeculae arranged densely and the connection between bone trabeculae increased in high-dose nourishing Yin and tonifying kidney recipe group. The expression levels of p-NF-κB/NF-κB, IkB, and MyD88 decreased, the levels of TNF-α, IL-6, IL-1β, IL-8, IL-10 decreased significantly, and the levels of Wnt-3α, LRP-5, β-catenin increased significantly in both middle-dose group and high-dose group (P<0.05). Conclusion The treatment of diabetic rats with ovariectomized osteoporosis with nourishing Yin and tonifying kidney recipe significantly increases the number of osteoblasts and bone mass possibly through up-regulation of Wnt/β-catenin signaling pathway.

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