补肾化痰方调控去势骨质疏松大鼠骨脂分化改善绝经后骨丢失的机制

Mechanism of Bushen Huatan Prescription in Regulation of Osteogenesis and Adipogenic Differentiation and Improvement of Postmenopausal Bone Loss in Ovariectomized Rats with Osteoporosis

目的:探讨补肾化痰方维持去势骨质疏松模型大鼠成骨/成脂分化平衡防治绝经后骨质疏松症(PMOP)的作用机制。方法:6月龄雌性未孕SD大鼠75只,随机分为假手术组、模型组、阿托伐他汀组、利维爱组、补肾化痰组,除假手术组外,其余4组大鼠均切除双侧卵巢,假手术组切除卵巢周围相应体积的脂肪组织。术后第5周起给予对应药物灌胃,剂量分别为补肾化痰组9.4 g·kg^(-1)、阿托伐他汀组0.92 mg·kg^(-1)、利维爱组0.23 mg·kg^(-1),模型组和假手术组给予等容生理盐水,1次/d,持续8周。采用微计算机断层扫描技术(Micro CT)检测各组大鼠胫骨骨密度(BMD),苏木素-伊红(HE)染色观察各组大鼠骨髓脂肪组织(BMAT)相对面积,实时荧光定量聚合酶链式反应(Real-time PCR)、蛋白免疫印迹法(Western blot)检测骨组织Runt相关转录因子2(Runx2)、过氧化物酶体增殖物激活受体γ(PPARγ)、瘦素(LPN)、瘦素受体(OBR)表达量。结果:与假手术组比较,模型组大鼠BMD明显下降(P<0.05),BMAT相对面积明显升高(P<0.05),LPN、OBR、Runx2表达明显下降(P<0.05),PPARγ明显升高(P<0.05)。与模型组比较,阿托伐他汀组、利维爱组、补肾化痰组BMD明显升高(P<0.05),BMAT相对面积明显下降(P<0.05),LPN、OBR、Runx2表达明显上升(P<0.05),PPARγ明显下降(P<0.05)。结论:补肾化痰方可能通过上调骨组织中LPN及OBR的表达,维持去势骨质疏松大鼠的骨脂分化平衡,从而减少绝经后骨丢失,发挥防治PMOP的作用。

Objective:To explore the underlying mechanism of Bushen Huatan prescription in alleviating postmenopausal osteoporosis(PMOP)by maintaining the balance of osteogenesis and adipogenic differentiation in ovariectomized rats with osteoporosis.Method:Seventy-five 6-month-old non-pregnant female SD rats were randomly divided into sham-operation group,model group,atorvastatin group,liviol group,and Bushen Huatan prescription group.Bilateral ovaries were removed in the four groups except the sham-operation group,while only the same mass of adipose tissue around the ovaries was removed in the shamoperation group.On the 5thweek after surgery,drugs were consecutively administrated for 8 weeks.Rats in the Bushen Huatan prescription group received 9.4 mg·kg^(-1) of the prescription,rats in the atorvastatin group received 0.92 mg·kg^(-1) of atorvastatin,rats in the Liviol group received 0.23 mg·kg^(-1) of liviol,and rats in the model group and the sham-operation group received saline once a day.Micro-computed tomography(Micro CT)was used to detect bone mineral density(BMD)of rat tibia in each group.Hematoxylin-eosin(HE)staining was used to detect the relative area of rat bone marrow adipose tissue(BMAT)in each group.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)and Western blot were used to detect the relative expression levels of Runt-related transcription factor 2(Runx2),peroxisome proliferator-activated receptor(PPARγ),leptin(LPN),and leptin receptor(OBR)in bone tissues.Result:As compared with the sham operation group,the BMD of rats in the model group decreased(P<0.05),while the relative area of BMAT increased(P<0.05).In addition,the expression levels of LPN,OBR,and Runx2 decreased in the model group(P<0.05),while the level of PPARγincreased(P<0.05).As compared with the model group,the BMD of rats in the atorvastatin group,the Livial group,and the Bushen Huatan prescription group increased(P<0.05),and the relative area of BMAT decreased(P<0.05).The expression levels of LPN,OBR,and Runx2 in these groups increased(P<0.05),while the expression level of PPARγdecreased(P<0.05).Conclusion:Bushen Huatan prescription plays the anti-osteoporosis role in the rat model of PMOP through up-regulating LPN and OBR in bone tissues and maintaining the balance of osteogenesis and adipogenic differentiation,thereby reducing postmenopausal bone loss and playing a role in the prevention and treatment of PMOP.