基于活性成分探讨山茱萸酒制前后干预绝经后骨质疏松症大鼠的动态药效学变化及其机制

Dynamic Pharmacodynamic Changes and Mechanism of Corni Fructus Before and After Wine-processed in Postmenopausal Osteoporosis Rats Based on Active Components

目的 比较研究山茱萸和酒茱萸中5-羟甲基糠醛(5-hydroxymethylfurfural,5-HMF)、环烯醚萜苷(莫诺苷、马钱苷、獐牙菜苷)、没食子酸5种活性成分含量差异,并探讨酒制前后山茱萸干预绝经后骨质疏松症(postmenopausal osteoporosis,PMOP)模型大鼠术后30,60,90 d的动态药效变化及其机制。方法 采用Hypersil C_(18)色谱柱(4.6 mm×250 mm,5μm),以乙腈(A)-0.3%磷酸溶液(B)为流动相,梯度洗脱,流速0.6 mL·min^(–1),柱温30℃,检测波长:240 nm(环烯醚萜苷),274 nm(5-HMF、没食子酸);将雌性SD大鼠随机分为假手术组,模型组,山茱萸/酒茱萸高、低剂量组,阳性药物组,除假手术组外均采用去卵巢法造模,分别在术后30,60,90 d,检测大鼠骨密度(bone mineral density,BMD)值、血清中的雌二醇(E_2)、钙(Ca)、磷(P)含量及骨组织显微结构的动态变化。结果 酒茱萸中5种活性成分总含量、5-HMF、没食子酸的含量均高于山茱萸;与模型组比较,酒茱萸高剂量组可升高模型大鼠术后30,60 d的E_2、Ca及P的含量(P<0.05),且术后60 d还可升高其BMD值(P<0.05);酒制前后山茱萸高剂量组均可升高模型大鼠术后90 d的BMD值和E_2、Ca及P含量(P<0.05),且酒茱萸高剂量组还可减少骨小梁的断裂及骨髓腔中脂肪细胞的含量。结论 酒制前后山茱萸均可改善PMOP,并以高剂量为优;且酒茱萸可早期干预该症的发生发展。其机制可能与升高E_2水平,减缓骨中Ca、P的丢失,调节骨代谢,延缓BMD值下降有关。酒茱萸中活性成分5-HMF、没食子酸的含量高于山茱萸可能是其药效较优的主要因素之一。

OBJECTIVE To compare the differences of five active components in Corni Fructus and wine-processed Corni Fructus: 5-hydroxymethyl furfural(5-HMF), iridoid glycosides(monoside, loganin, swertioside), gallic acid, and to explore the dynamic pharmacodynamic changes and mechanism on postmenopausal osteoporosis(PMOP) model rats of Corni Fructus and wine-processed Corni Fructus on 30, 60, 90 d after operation. METHODS The analysis was performed on Hypersil C18 chromatography(4.6 mm×250 mm, 5 μm), with the gradient elution of acetonitrile(A)-0.3% phosphoric acid(B) solution at flow rate of 0.6 mL·min^(-1). The column temperature was 30 ℃ and the detection wavelength was set at 240 nm (iridoid glycoside), 274 nm (5-HMF, gallic acid). The female SD rats were randomly divided into sham group, model group, low-dose and high-dose of Corni Fructus and wine-processed Corni Fructus groups, and all of them were emasculated and modeled except the sham group. On 30, 60 and 90 d after operation, the dynamic changes of bone mineral density(BMD), serum estradiol(E2), calcium(Ca), phosphorus(P) and microstructure of bone tissue were measured. RESULTS The total content of five active components, 5-HMF and gallic acid in wine-processed Corni Fructus was higher than that of Corni Fructus. Compared with the model group, the high-dose group of wine-processed Corni Fructus could increase the levels of serum E2, Ca and P on the 30 and 60 d after operation, the BMD could also be increased on the 60th day after operation(P<0.05). On the 90 d after operation, both the high-dose of Corni Fructus and wine-processed Corni Fructus groups could increase the BMD, serum E2, Ca and P levels of the model rats(P<0.05), the high-dose wine-processed Corni Fructus group could also reduce the fracture of bone trabeculae and the content of adipocytes in the bone marrow cavity. CONCLUSION Corni Fructus and wine-processed Corni Fructus can alleviate PMOP,high-dose is better;wine-processed Corni Fructus can intervene the occurrence and development of the disease in the early stage.The mechanism may be related to increasing E2 level,slowing down the loss of Ca and P in bone,regulating bone metabolism and delaying the decline of bone density,the higher 5-HMF and gallic acid than that of Corni Fructus may be one of the main factors for the better efficacy of wine-processed Corni Fructus.