桑叶中芦丁含量的动态变化研究

目的：研究不同时期桑叶中芦丁的含量。方法 Kromasil C18柱（4.6 mm ×250 mm,5μm）;流动相：甲醇-0.5％磷酸水溶液（梯度洗脱）,检测波长：358 nm;柱温：30℃;结果芦丁在0.1262~2.02μg范围内有良好的线性关系,r＝0.9999,平均回收率为97.88％,桑叶中芦丁含量最高在霜降前后。结论初步表明了桑叶中芦丁含量随季节变化的规律。     

Rutin determination in pharmaceutical formulations using a carbon paste electrode modified with poly(vinylpyrrolidone)

A carbon paste electrode modified with poly(vinylpyrrolidone) (PVP) was developed for the determination of rutin. PVP enhances the adsorption of rutin on the electrode surface due to the presence of hydrogen bonding between the imide group in PVP and the hydroxyl group in rutin. The current responses in cyclic and linear sweep voltammetric experiments were investigated and an oxidation peak was observed at +0.87 V vs. Ag/AgCl. Several parameters were investigated to evaluate the performance of the modified electrode. The best analytical response was obtained employing (75:15:10%, w/w/w) graphite powder:Nujol:PVP, with an accumulation time of 10 min, scan rate of 100 mV s(-1) and 0.1M phosphate buffer solution (pH 6.0) as supporting electrolyte. The analytical curve was linear for rutin concentrations of 3.9 x 10(-7) to 1.3 x 10(-5)M (r=0.9991) and the detection limit was 1.5 x 10(-7)M. The recovery of rutin from pharmaceutical samples ranged from 98.3 to 101.7% and the relative standard deviation was 3.3% for a solution containing 7.7 x 10(-6)M rutin (n=5). This electrode was successfully applied to the determination of rutin in pharmaceutical formulations. The results obtained using the proposed modified carbon paste electrode and those obtained by the standard method are in agreement at the 95% confidence level.     

炎症微环境下芦丁对牙周膜干细胞成骨分化能力的影响

目的 研究芦丁（rutin）对炎症微环境下牙周膜干细胞成骨分化能力的影响。方法 采用有限稀释法分离纯化获得牙周膜干细胞,采用流式细胞术鉴定牙周膜干细胞。以脂多糖（lipopolysaccharide, LPS）刺激牙周膜干细胞,建立体外炎症模型。实验分为4组,第1组使用α-MEM培养基培养牙周膜干细胞,第2组使用含有脂多糖的α-MEM培养基培养牙周膜干细胞,第3组在含有脂多糖的α-MEM培养基加入芦丁培养牙周膜干细胞,第4组使用含有芦丁的α-MEM培养基培养牙周膜干细胞。通过碱性磷酸酶染色、碱性磷酸酶活性测试、茜素红染色、RT-PCR以及蛋白免疫印迹等方法检测成骨分化能力的改变。采用SPSS 17.0软件包对数据进行统计分析。结果 CCK-8和碱性磷酸酶活性测试结果显示,10 μmol/L芦丁对炎症状态下牙周干细胞增殖和成骨分化作用最明显。碱性磷酸酶染色和茜素红染色结果显示,10 μmol/L芦丁可以改善炎症微环境下牙周膜干细胞的成骨分化能力。RT-PCR、蛋白免疫印迹结果显示,芦丁可以增强炎症状态下COL1、ALP、RUNX2等成骨基因和成骨蛋白的表达。结论 芦丁可以增强炎症微环境下牙周膜干细胞的成骨分化能力。     

Effects of Watercress Containing Rutin and Rutin Alone on the Proliferation and Osteogenic Differentiation of Human Osteoblast-like MG-63 Cells

Most known osteoporosis medicines are effective for bone resorption, and so there is an increasing demand for medicines that stimulate bone formation. Watercress (N. officinale R. Br.) is widely used as a salad green and herbal remedy. This study analyzed a watercress extract using ultra-performance liquid chromatography/mass spectrometry, and identified a rutin as one of its major constituents. Osteogenic-related assays were used to compare the effects of watercress containing rutin (WCR) and rutin alone on the proliferation and differentiation of human osteoblast-like MG-63 cells. The reported data are expressed as percentages relative to the control value (medium alone; assigned as 100%). WCR increased cell proliferation to 125.0±4.0% (mean±SD), as assessed using a cell viability assay, and increased the activity of alkaline phosphatase, an early differentiation marker, to 222.3±33.8%. In addition, WCR increased the expression of collagen type I, another early differentiation marker, to 149.2±2.8%, and increased the degree of mineralization, a marker of the late process of differentiation, to 122.9±3.9%. Rutin alone also increased the activity of ALP (to 154.4±12.2%), the expression of collagen type I (to 126.6±6.2%), and the degree of mineralization (to 112.3±5.0%). Daidzein, which is reported to stimulate bone formation, was used as a positive control; the effects of WCR on proliferation and differentiation were significantly greater than those of daidzein. These results indicate that WCR and rutin can both induce bone formation via the differentiation of MG-63 cells. This is the first study demonstrating the effectiveness of either WCR or rutin as an osteoblast stimulant.     

畲药食凉茶中三种黄酮类成分含量测定的方法学研究

目的：建立同时测定畲药食凉茶中三种黄酮类成分含量的方法。方法：采用高效液相色谱法测定,色谱柱为Agilent Zorbax SB-C18色谱柱（250 mm×4.6 mm,5μm）;流动相为乙腈-0.1%磷酸溶液,梯度洗脱;流速为1.0 ml·min^-1,柱温30℃;检测波长360 nm。结果：芦丁、槲皮素和山柰酚的线性范围分别为0.040 9-1.637 0 mg·ml^-1（r=0.999 2）,0.44-88.00μg·ml-1（r=0.999 8）,0.41-77.63μg·m^l-1（r=0.999 2）;平均加样回收率分别为99.35%（RSD=1.64%）,101.14%（RSD=1.88%）,99.69%（RSD=1.92%）。结论：该方法简便易行、快速准确、重复性好,可用于同时测定食凉茶中的芦丁、槲皮素和山柰酚的含量。     

Protective effect of rutin on testicular ischemia-reperfusion injury

Purpose

The pathophysiology of testicular torsion-detorsion is an ischemia-reperfusion injury caused by overgeneration of reactive oxygen species (ROS). This study aimed to investigate the effect of rutin, a well-known antioxidant, on testicular ischemia-reperfusion injury.

Methods

Sixty male Sprague-Dawley rats were randomly divided into 3 groups, each containing 20 rats. Rats in the control group underwent a sham operation of the left testis. In the torsion-detorsion group, the left testis was rotated 720° for 2 hours. Rats in the treatment group received the same surgical procedure as the torsion-detorsion group, but rutin was administered intravenously at the time of detorsion. Bilateral orchiectomy was performed on half of the rats in each experimental group at 4 hours after detorsion for measurement of malondialdehyde, an indicator of intratesticular ROS content, and for evaluation of superoxide dismutase and catalase, which are endogenous antioxidant enzymes. Orchiectomy was performed on the remaining rats at 3 months after detorsion for analysis of testicular spermatogenesis.

Results

Unilateral testicular torsion-detorsion caused a significant increase in malondialdehyde level and caused significant decreases in superoxide dismutase, catalase activities, and spermatogenesis in ipsilateral testes. The rats treated with rutin had a significant decrease in malondialdehyde level and had significant increases in superoxide dismutase, catalase activities, and spermatogenesis in ipsilateral testes, compared with torsion-detorsion group.

Conclusions

Rutin protects testes from ischemia-reperfusion injury. The protective effect of rutin may be caused by scavenging ROS by increasing superoxide dismutase and catalase activities.     

测定新疆阔克阿木提果皮中绿原酸和芦丁的含量

目的:建立采用超声波提取,高效液相色谱法测定新疆阔克阿木提果皮中绿原酸和芦丁的含量的方法。方法:色谱柱:安捷伦HC-C18柱(250mm×4.6mm,5μm);流动相体积比50∶50的甲醇-0.4%磷酸溶液;检测波长为360nm;柱温为30℃;流速为1.0ml/min低压梯度洗脱。结果:绿原酸和芦丁分别在12.5~150μg/ml(r=0.9998)和12.5~150μg/ml(r=0.9998)浓度范围内呈良好的线性关系,平均回收率分别为101.08%(RSD=1.38%,n=5)和99.56%(RSD=1.17%,n=5)。结论:该方法简便快速、重现性好、准确可靠,适用于新疆阔克阿木提果皮中绿原酸和芦丁的含量测定。     

不同品种丁香叶中芦丁及丁香苦苷的含量测定

目的:建立多波长高效液相色谱法(HPLC)同时测定不同品种丁香叶中芦丁及丁香苦苷的含量。方法:采用AgilentZORBAXEclipseXDB-C18色谱柱(4.6mm×250mm,5μm),流动相为乙腈-0.1%磷酸水(20∶80),流速为1.0mL/min,定性检测波长200～400nm,定量检测波长255nm和225nm,柱温为室温;利用化学计量学中的光谱相关色谱法分析确认定量色谱峰。结果:芦丁和丁香苦苷在20min内被很好地分离,经对照品比较及光谱分析得到确认。芦丁、丁香苦苷分别在0.125～2.0μg和0.625～10.0μg范围内与色谱峰面积的线性关系良好(r=0.9999)。平均回收率分别为99.9%、99.4%,RSD分别为1.4%、1.1%。结论:不同品种丁香叶中芦丁和丁香苦苷的含量有较大的差别,建立的含量测定方法可用于丁香叶中芦丁及丁香苦苷的同时定量分析。     

高效液相色谱法同时测定不同产地、不同炮制时间连翘和连翘叶中5种活性成分的含量

目的:采用HPLC法测定不同产地、不同炮制时间连翘和连翘叶中芦丁、连翘酯苷A、(+)松脂素-β-D-葡萄糖苷、(+)表松脂素-β-D-葡萄糖苷、连翘苷。方法:色谱柱:Waters Symmetry C18(4.6 mm×250 mm,5μm),柱温25℃,流动相乙腈-0.4%冰醋酸溶液,梯度洗脱,流速1 mL.min-1,检测波长277 nm。结果:芦丁、连翘酯苷A、(+)松脂素-β-D-葡萄糖苷、(+)表松脂素-β-D-葡萄糖苷、连翘苷在检测的范围内线性关系良好,r2均大于0.999 7,方法回收率在97.69%～100.30%之间。结论:此方法简便、准确,快速,为全面评价连翘和连翘叶的质量提供可靠的分析方法。     

紫外分光光度法测定金莲花口服液含量

目的建立金莲花口服液中黄酮类物质的含量测定方法。方法采用紫外分光光度法,通过紫外扫描确定最大吸收波长,在最大吸收波长处以芦丁为对照品测量吸光度制定标准曲线,并进行样品溶液稳定性试验、重现性试验、精密度试验、回收率试验等。结果精密度试验所测得的吸光度分别为:0.235、0.234、0.234、0.234、0.233,相对标准偏差(RSD)=0.30%,数据表明该试验具有良好的精密度;稳定性试验测定在0、5、10、15、20、25min时吸光度分别为0.249、0.249、0.248、0.245、0.245、0.244,RSD=0.83%,数据表明在25min内对待测物进行吸光度的测定是稳定的;重现性试验在500nm波长处测定吸光度为:0.412、0.411、0.413、0.407、0.407,RSD=0.69%,表示重现性良好;回收率试验测定的平均回收率为98.5%,RSD=0.56%;样品含量的测定测定的3种批号样本的平均含量分别为0.334mg/ml、0.412mg/ml、0.375mg/ml。结论紫外分光光度法简单方便、准确可靠,可以用来测定金莲花口服液中黄酮类物质的含量。