刺芒柄花素对去势骨质疏松大鼠P38MAPK/MMP-9信号通路的影响

目的:观察刺芒柄花素（formononetin，FOR）对大鼠去势骨质疏松的干预作用，探讨其防治骨质疏松的机制。方法:采用切除雌性大鼠双侧卵巢法复制骨质疏松大鼠模型，按随机对照分组原则将60只清洁级SD雌性大鼠分为对照组、骨质疏松模型组、雌二醇组(50 μg/kg)及刺芒柄花素高、中、低剂量组(160、80、40 mg/kg)，每组10只，除对照组外，其他组于造模后一周给予雌二醇、刺芒柄花素治疗，每天1次，连续12周，末次给药后24 h，取大鼠血清检测酒石酸酸性磷酸酶（TRACP）、Ca2+、Mg2+等指标变化；取大鼠左侧股骨采用RT-PCR检测基质金属蛋白酶-9（MMP-9）、TNF-α、IL-6 mRNA表达；Western blot法检测骨组织总P38MAPK、磷酸化P38MAPK、MMP-9、TNF-α、IL-6蛋白表达；取右侧股骨进行骨密度(bone mineral density，BMD)测定。结果:与骨质疏松模型组比较，刺芒柄花素高、中剂量组可增加骨质疏松大鼠骨密度，降低骨质疏松大鼠血清中TRACP含量，升高血清 Mg2+、Ca2+含量，使骨组织MMP-9、TNF-α、IL-6 mRNA和蛋白表达下调;磷酸化P38MAPK mRNA蛋白表达上调。结论:刺芒柄花素对去势骨质疏松大鼠有较好的防治作用，其机制可能与抑制骨组织MMP-9的合成和分泌、破骨细胞的骨吸收及调控P38MAPK/MMP-9 信号通路有关。

Effects of formononetin on P38MAPK/MMP-9 signaling pathway in ovariectomized rats

AIM: To observe the intervention effects of formononetin (FOR) on castration osteoporosis in rats, and the mechanism of osteoporosis prevention. METHODS: The bilateral ovariectomy was performed on female rats to induce osteoporosis rat model. Sixty female SD rats were divided into normal control group, osteoporosis model group, estradiol group (50 μg/kg) and formononetin in high, medium and low dose group (160, 80, 40 mg/kg), 10 rats in each group. except the control group,All groups were administered with estradiol or formononetin treatment,(1 time a day) for 12 weeks one week after operation;rats serum tartrate resistant acid phosphatase (TRACP), change Ca ²⁺, Mg ²⁺ and other indicators were detected 24 h after the last injection;the left femur was observed to detect MMP-9 and TNF-alfa, IL-6 mRNA expression by RT-PCR;Western-blot was used to detect P38MAPK, total bone tissue phosphorylation of P38MAPK, MMP-9,TNF-alfa, IL-6 protein; the right femoral bone was observed for density (bone mineral densit Y, BMD) determination. RESULTS: Compared with osteoporosis model rats, formononetin high dose group can increase the bone density of rats with osteoporosis, lower TRACP content in serum of rats with osteoporosis, and increase serum Mg ²⁺, Ca ²⁺ content, reduce MMP-9, TNF-alfa and the bone IL-6, mRNA expression regulated by phosphorylation; P38MAPK mRNA protein expression. CONCLUSION: Formononetin has a good preventive and therapeutic effect on ovariectomized osteoporotic rats, and its mechanism may be related to inhibiting MMP-9 synthesis and secretion, osteoclast bone resorption and regulating P38-MAPK/MMP-9 signaling pathway.