炙淫羊藿炮制工艺及抗骨质疏松作用

Study on the processing technology and anti-osteoporosis effect of roasted Epimedium

目的优选炙淫羊藿的炮制工艺参数,并基于斑马鱼模型评价炙淫羊藿抗骨质疏松的作用。方法以总黄酮醇苷(朝藿定A、朝藿定B、朝藿定C和淫羊藿苷)及宝藿苷Ⅰ质量分数为评价指标,通过设计单因素试验和正交试验,优选炙淫羊藿的炮制温度、炮制时间和转速,确定最佳炮制工艺参数。采用优选的炙淫羊藿进行斑马鱼的急性毒性试验,确定炙淫羊藿水提液的最大耐受浓度(MTC);采用4 dpf的野生型AB品系斑马鱼,以25μmol/L地塞米松造模,评价炙淫羊藿水提液对骨质疏松症的治疗作用。结果炮制温度为主要影响因素,炮制时间及转速对炮制结果影响不显著。优选的炙淫羊藿最佳炮制工艺参数为:炮制温度160℃、炮制时间10 min、转速12 r/min。炙淫羊藿水提液在24、48、72、96、120 h的MTC分别为2、1、1、0.5、0.5 mg/mL;与模型组比较,各给药组的斑马鱼脊椎节个数和前三节脊椎平均荧光强度差异均有统计学意义(P<0.05和P<0.01)。结论优选的炙淫羊藿炮制工艺重复性好、稳定可行;炙淫羊藿水提液可有效改善地塞米松引起的斑马鱼骨质疏松症,且存在一定的量效关系。

Objectiv To optimize the processing parameters of roasted Epimedium and evaluate the antiosteoporosis effect of roasted Epimedium based on a zebrafish model.Methods Using the total flavonol glycosides(chaohuoding A,chaohuoding B,chaohuoding C,and Epimedium glycoside)and baohuoside I as the evaluation index,single factor experiments and orthogonal experiments were designed to optimize the processing temperature,time,and speed of roasted Epimedium,and determine the optimal processing technology.Acute toxicity tests were conducted on zebrafish using optimized roasted Epimedium to determine the maximum tolerable concentration(MTC)of the water extract of roasted Epimedium.Using wild-type AB strain zebrafish with 4 days after fertilization,a 25μmol/L dexamethasone model was used to evaluate the therapeutic effect of roasted Epimedium on osteoporosis.Results Processing temperature was the main influencing factor,while time and speed had no significant impact on the processing results.The optimal processing technology for roasted Epimedium was 160℃,10 min and 12 r/min.The MTC of the water extract of roasted Epimedium at 24,48,72,96,and 120 h were 2,1,1,0.5 and 0.5 mg/mL,respectively.Compared with the model group,the number of vertebrae and the average fluorescence intensity of the first three vertebrae in each administration group showed statistical differences to varying degrees(P<0.05 and P<0.01).Conclusion The optimal processing technology for roasted Epimedium has good reproducibility,stability,and feasibility.The water extract of Epimedium can effectively improve zebrafish osteoporosis caused by dexamethasone,and there is a certain dose-response relationship.