补肾活血中药对骨质疏松大鼠Wnt信号的影响

Effect of enriching kidney and promoting blood flow TCM on Wnt signal in osteoporotic rats

目的探讨补肾活血方对卵巢摘除骨质疏松大鼠Wnt通路β-Catenin mRNA表达的影响。方法 SPF级250±20 g雌性SD大鼠40只,随机分为假手术组(sham group)、模型组(model group)、雌激素药物治疗组(E group)、中药组(TCM group),每组10只。对假手术组大鼠施行假手术,其余4组行双侧卵巢摘除术,术后30天进行相应的给药实验,假手术组和模型组给予灌服等量的生理盐水。12周后测定各组大鼠血清中I型胶原羧基端肽(β-Crosslaps)、I型前胶原氨基端延长肽(P1NP)含量;以双能X线骨密度测定仪测定大鼠的骨密度;用RT-PCR检测右侧股骨髓LRP5、Wnt2、β-Catenin mRNA表达。结果model组动物骨密度值(0.137±0.013 g/cm2)、P1NP浓度(4.388±0.400 ng/ml)、β-Catenin mRNA表达(1.05±0.18)、LRP5mRNA表达(0.96±0.24)、Wnt2 mRNA表达(0.53±0.11)。与模型组比较,sham组及TCM组BMD(0.168±0.008,0.173±0.008)显著增高(P<0.05),血清β-Crosslaps(0.575±0.068,0.618±0.119)、LRP5 mRNA表达(1.63±0.51,2.630.22)显著增高(P<0.05),sham、E、TCM组Wnt2mRNA表达(1.03±0.02,1.23±0.12,1.58±0.20)及β-Catenin mRNA表达(1.95±0.16,1.62±0.17,2.12±0.20)显著增高(P<0.05);与sham组比较,TCM组β-Crosslaps、BMD(0.618±0.119,0.173±0.008)没有差异(P>0.05),P1NP、Wnt2、LRP5 mRNA表达(6.310±0.990,1.58±0.20,2.63±0.22)显著增高(P<0.05);与E组比较,TCM组PINP、BMD、Wnt2、β-catenin、LRP5显著增高(P<0.05)。结论补肾活血中药通过调节Wnt信号促进去卵巢术后骨质疏松大鼠成骨细胞增殖。

Objective To investigate the effect of traditional Chinese medicine（ TCM） enriching kidney and promoting blood flowreceipt on β-catenin mRNA in Wnt signal pathway in osteoporosis rat model. Methods Forty 250 ± 20 g female SPF grade SD rats were randomly divided into 4 groups： the sham operation group（ sham group）,the model group,the estradiol treatment group（ E group）,and the TCMenriching kidney and promoting blood flowtreatment group（ TCMgroup）. Ten rats were in each group. Rats in the sham group were sham operated,and rats in the other groups were bilateral ovariectomized. The rats received corresponding treatment for 30 days after operation. After 12 weeks,bone mineral density（ BMD） of all rats was measured using dual-energy X-ray absorptiometry. Serum concentration of type I collagen carboxy-terminal peptide（ β-Crosslaps） and aminoterminal propeptide of type 1 procollagen（ P1NP） were determined. The expression of Wnt2,LRP5,and β-catenin in the right femoral bone marrowwas measured. Re sults In the model group,BMD was 0. 137 ± 0. 013 g / cm^2,and serum concentration of P1 NP was 4. 388 ± 0. 400 ng / ml. The expression of β-Catenin mRNA,LRP5 mRNA,and Wnt2 mRNA was 1. 05 ± 0. 18,0. 96 ±0. 24,and 0. 53 ± 0. 11,respectively. Compared to the model group,BMD in sham（ 0. 168 ± 0. 008） and TCMgroup（ 0. 173 ±0. 008） increased significantly（ P〈0. 05）,and β-Crosslaps（ 0. 575 ± 0. 068,0. 618 ± 0. 119） and the expression of LRP5 mRNA（ 1. 63 ± 0. 51,2. 63 ± 0. 22） increased significantly（ P〈0. 05）. The expression of Wnt2 mRNA and β-Catenin mRNA in sham,E,and TCMgroup increased obviously compared to the model group. P1 NP and the expression of Wnt2 and LRP5 mRNA in TCMgroup increased significantly compared to the sham group（ P〈0. 05）,but β-Crosslaps and BMD between TCMand the sham group were no difference. Compared to E group,PINP,BMD,Wnt2,β-catenin,and LRP5 in TCMgroup increased significantly（ P〈0. 05）. Conclusion TCMenriching kidney and promoting blood flowpromotes osteoblast proliferation by regulating Wnt signal in ovariectomized osteoporotic rats.