目的:探讨葛根素(puerarin, Pue)治疗绝经后骨质疏松症(postmenopausal osteoporosis, PMOP)大鼠的效果及作用机制。方法:将雌性SD大鼠分为假手术组(SHAM组)、模型组(PMOP组)、葛根素低剂量组(Pue-L组)和葛根素高剂量组(Pue-H组)。SHAM组取卵巢周围部分脂肪,其余各组切除双侧卵巢,术后4周检测骨矿物质密度(bone mineral density,BMD)和苏木精-伊红(HE)染色法观察股骨组织,判断造模是否成功。造模成功后Pue-L组和Pue-H组大鼠皮下注射不同浓度葛根素注射液,SHAM组和PMOP组注射等量生理盐水。给药8周后检测大鼠BMD;用显微CT(Micro-CT)扫描观察骨结构并计算相关参数;HE染色观察大鼠股骨病理改变;Western Blot(WB)法检测JAK2/STAT3信号通路蛋白表达水平。结果:(1)Pue可显著提高PMOP大鼠BMD。(2)Micro-CT分析显示,Pue可改善PMOP大鼠骨微结构。(3)HE染色显示Pue治疗后PMOP大鼠骨量丢失减少、骨小梁数量与密度增加,病理改变有所恢复。(4)WB检测结果:与SHAM组相比,PMOP组p-JAK2、p-STAT3蛋白表达水平升高(P＜0.001);与PMOP组比较,Pue-L组和Pue-H组p-JAK2、p-STAT3蛋白表达水平降低(P＜0.001)。结论:葛根素可有效治疗PMOP大鼠,并可能通过抑制JAK2/STAT3通路的激活发挥抗骨质疏松作用。
Objective: To investigate the effect and mechanism of puerarin (Pue) in the treatment of postmenopausal osteoporosis (PMOP) in rats. Methods: Female SD rats were divided into sham operation group (SHAM group), model group (PMOP group), low-dose puerarin group (Pue-L group) and high-dose puerarin group (Pue-H group). In the SHAM group, partial fat around the ovary was removed, and bilateral ovaries were removed in the other groups. Bone mineral density (BMD) and hematoxylin-eosin (HE) staining were used to observe the femoral tissue at 4 weeks after operation to determine whether the model was successful. After successful modeling, rats in Pue-L group and Pue-H group were subcutaneously injected with different concentrations of puerarin injection, and rats in SHAM group and PMOP group were injected with the same amount of normal saline. BMD of rats was detected after 8 weeks of administration. The bone structure was observed by micro-CT scanning and the related parameters were calculated. HE staining was used to observe the pathological changes of rat femur. The protein expression level of JAK2/STAT3 signaling pathway was detected by Western Blot (WB). Results: (1) Pue significantly increased BMD in PMOP rats. (2) Micro-CT analysis showed that Pue could improve the bone microstructure of PMOP rats. (3) HE staining showed that after Pue treatment, the bone loss of PMOP rats decreased, the number and density of trabecular bone increased, and the pathological changes were restored. (4) WB results showed that compared with SHAM group, the expression levels of p-JAK2 and p-STAT3 protein in PMOP group were increased (P＜0.001). Compared with PMOP group, the expression levels of p-JAK2 and p-STAT3 protein in Pue-L group and Pue-H group were decreased (P＜0.001). Conclusion: Puerarin can effectively treat osteoporosis in PMOP rats, which may play an anti-osteoporosis role by inhibiting the activation of JAK2/STAT3 pathway.
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