女贞子炮制前后5-羟甲基糠醛的含量变化

目的 研究女贞子炮制前后5-羟甲基糠醛(5-hydroxymethyl furfural,5-HMF)的含量变化.方法 制备不同炮制条件的女贞子,采用HPLC法测定女贞子生品及炮制品中5-HMF的含量.结果 女贞子生品未检测到5-HMF,炮制后产生5-HMF,不同炮制品中的含量依次为:酒炖品＞清蒸品＞酒蒸品,且随蒸制时间延长含量增加.结论 首次建立了HPLC测定女贞子中5-HMF含量的方法,该法稳定可靠,可用于其质量控制;蒸制方法和蒸制时间均对5-HMF的含量变化有影响,这种改变是否与炮制增效有关,值得进一步研究.     

女贞子中三萜类化合物研究

目的:研究木犀科植物女贞子的三萜类成分。方法:用溶剂提取、萃取、硅胶正相柱色谱、Sephadex LH-20柱色谱及结晶法进行化学成分的分离纯化,通过理化性质和波谱分析鉴定结构。结果:从女贞子中分离鉴定了13个化合物,分别为:齐墩果酸甲酯(1)、3-O-乙酰熊果酸(2)、3-羰基齐墩果酸(3)、19α-羟基熊果酸(4)、羽扇豆醇(5)、3-O-乙酰齐墩果酸(6)、齐墩果酸(7)、白桦脂醇(8)、熊果酸(9)、fouquierol(10)、19α-羟基-3-O-乙酰熊果酸(11)、(Z)-马斯里酸-3-O-对香豆酸酯(12)、(E)-马斯里酸-3-O-对香豆酸酯(13)。结论:其中,化合物1～4为首次从该植物中分离得到。     

女贞子炮制过程中2种三萜类成分及水溶性低分子糖含量变化规律研究

目的:研究不同炮制条件对女贞子中三萜类成分齐墩果酸、熊果酸及低分子糖含量的影响,探讨女贞子酒制过程中齐墩果酸、熊果酸及低分子糖含量的动态变化规律。方法:对女贞子生品、清蒸品、酒蒸品及不同加热时间酒炖品中齐墩果酸、熊果酸和低分子糖含量进行测定比较。结果:女贞子经不同方法炮制后齐墩果酸和熊果酸含量无明显变化,而水溶性低分子糖含量增加,且随着炖制时间的延长,炮制品中低分子糖含量逐渐增加。结论:炮制对女贞子中齐墩果酸和熊果酸含量无明显影响,对低分子糖含量有显著影响。     

响应面法优化女贞子中总三萜的提取工艺研究

目的采用星点设计-响应面法优选女贞子中总三萜的乙醇回流提取工艺。方法在单因素考察基础上,利用三因素三水平响应面法研究乙醇浓度、提取时间、溶媒比对女贞子总三萜得率的影响。结果确定最优提取工艺为12倍量67%乙醇提取2次,每次1.70 h;女贞子总三萜提取量为57.62 mg/g,与预测值的偏差为2.89%。结论星点设计-响应面法优化的女贞子总三萜提取工艺方法简单,精密度高,可预测性较优。     

Hepatoprotective action of an oleanolic acid-enriched extract of Ligustrum lucidum fruits is mediated through an enhancement on hepatic glutathione regeneration capacity in mice.

The fruits of Ligustrum lucidum Ait. (FLL) were fractionated into petroleum ether (FLL-Pe), chloroform (FLL-Ch), butanol (FLL-Bu) and aqueous (FLL-Aq) fractions, of which FLL-Ch and FLL-Bu were found to be enriched with oleanolic acid (OLA). The in vivo antioxidant activities of various FLL fractions and OLA were assessed by examining the effect on carbon tetrachloride (CCl(4))-induced hepatotoxicity in mice. Pretreatment of animals with various FLL fractions could protect against CCl(4)-induced hepatotoxicity to a varying degree, with OLA-enriched FLL-Bu and FLL-Ch being more potent. However, a mortality rate of 60% was observed in the FLL-Ch pretreated and CCl(4)-intoxicated mice. OLA pretreatment also produced a dose-dependent protection against CCl(4) hepatotoxicity. The hepatoprotection afforded by FLL-Bu or OLA pretreatment was associated with an enhancement of hepatic-glutathione regeneration capacity (GRC). In contrast, the inability of FLL-Aq pretreatment to enhance hepatic GRC resulted in a failure to prevent CCl(4)-induced hepatic injury. The results suggest that the hepatoprotective action afforded by OLA-enriched FLL-Bu or OLA pretreatment may be mainly mediated by the enhancement of hepatic GRC, particularly under conditions of CCl(4)-induced oxidative stress.     

甘肃产女贞子挥发油化学成分研究

 目的 研究甘肃产女贞子挥发油的化学成分,比较不同产地的女贞子挥发油药用成分。方法 采用常规水蒸汽蒸馏法对甘肃天水产女贞子提取挥发油,用气相色谱质谱联机进行分离鉴定。结果 分离出57种成分,鉴定出50种化合物,其峰面积相对含量约占挥发油总量的96.38%。结论 从本研究的结果知,药用成分主要是桉油精4.95%,苯甲醇2.50%,乙酸龙脑酯0.2%,三苯甲醇6.63%等;甘肃产女贞子与湖南产女贞子挥发油化学成分差距较大,其原因与生长条件、分离时的色谱柱有关。发现具有中等极性的SE-54色谱柱对极性较小的烷烃及分子量较大的醇醛类具有较好的分离效果。     

Protective effect of oleanolic acid on gentamicin induced nephrotoxicity in rats

Oleanolic acid is a molecule of current therapeutic interest. In the present study, oleanolic acid isolated from the cuticular epithelium of Viscum articulatum Burm. f. (Viscaceae) was investigated for its protective effects on gentamicin‐induced renal damage in rats. Nephrotoxicity was induced in rats by intraperitoneal injection of gentamicin at a dose of 100 mg/kg/day for 8 days. The effect of Oleanolic acid administered orally at doses 40, 60 and 80 mg/kg/day was assessed biochemically by determination of albumin, urea and creatinine in serum and urine samples and also through histopathological examination of the kidneys. Oleanolic acid protected the rat kidneys from gentamicin‐induced nephrotoxicity as evident from a decrease in the serum and urine levels of creatinine, albumin and urea. Oleanolic acid also protected the rat kidneys from histological alterations induced by gentamicin and also improved the glomerular filtration rate. Compared with an earlier report on intraperitoneal administration of oleanolic acid in paracetamol‐induced nephrotoxicity in rats, the data show that orally administered oleanolic acid also exerted a nephroprotective effect even in the case of a nephrotoxicant such as gentamicin, which directly deteriorates the kidney function without prior metabolism. Copyright © 2009 John Wiley & Sons, Ltd.     

女贞子提取物总三萜酸降血糖作用的实验研究

目的:观察女贞子三萜酸对实验性糖尿病大鼠的治疗作用。方法:建立大鼠糖尿病模型观察该药对空腹血糖、血清胰岛素、胆固醇、甘油三酯、糖化血清蛋白的影响,以及对正常大鼠糖耐量、血糖的影响。结果:女贞子三萜酸能明显降低血糖(P<0.01)和甘油三酯(P<0.01),改善糖耐量;对血清胰岛素、糖化血清蛋白、胆固醇等影响不明显。结论:女贞子三萜酸降血糖作用较强,对血酯具有一定的调节作用。     

Antidiabetic effect of oleanolic acid: a promising use of a traditional pharmacological agent

Diabetes mellitus (DM) is a metabolic disorder characterized by chronic hyperglycemia. Although the clear mechanisms of DM and insulin resistance are still to be cleared, it has been well documented that reactive oxygen species (ROS) play a pivotal role in DM and multiple types of insulin resistance. For the past few years, natural substances have been shown to have the potential to treatment DM. Attention has been especially focused on plants rich in triterpenoids, which generally show antioxidant and antiglycation effect. In our previous studies, it was shown that oleanolic acid (OA), a natural triterpenoid and an aglycone of many saponins, is a potent antioxidant acting as not only a free radical-scavenger through direct chemical reactions but also as a biological molecule, which may enhance the antioxidant defenses. The present study aimed to investigate the potential antidiabetic effect of OA. Oleanolic acid showed a significant blood glucose-lowering and weight-losing effect in diabetic animals induced by streptozotocin (STZ). In the insulin resistant model, it was also shown that OA may promote insulin signal transduction and inhibit oxidative stress-induced hepatic insulin resistance and gluconeogenesis, in which process the phosphorylation of ERK and the protective effect on mitochondrial function may be involved. These findings may significantly better the understanding of the pharmacological actions of OA and advance therapeutic approaches to DM.     

齐墩果酸脂质体在大鼠体内药代动力学的研究

目的:制备齐墩果酸脂质体,研究齐墩果酸脂质体在大鼠体内的药代动力学。方法:采用乙醇注入-探头超声法制备齐墩果酸脂质体;大鼠分别尾静脉注射齐墩果酸脂质体和齐墩果酸溶液剂,用反相高效液相色谱法测定不同时间血浆中齐墩果酸的浓度,采用3p97程序计算药代动力学参数,并进行统计学分析。结果:制得的脂质体透射电镜下为圆形或类圆形的小囊泡,平均粒径为(206.4±4.7)nm,包封率为(97.74±1.34)%;齐墩果酸脂质体和自制的溶液剂经大鼠尾静脉注射后,t1/2α分别为(4.22±1.86)和(0.80±0.34)min;t1/2β分别为(33.59±12.53)和(15.65±7.91)min;AUC分别为(240.13±23.62)和(168.31±18.47)μg/(mL.min),两种制剂的t1/2β和AUC0-t经方差分析后均存在显著性差异(P0.05),齐墩果酸脂质体显著增加了齐墩果酸的AUC,延长了其在血液循环中的时间。结论:制备的齐墩果酸脂质体包封率较高,粒度均匀;与非脂质体相比,齐墩果酸脂质体静脉注射给药后,药代动力学参数发生了明显的改变。     

HPLC法测定陕西女贞子中齐墩果酸与熊果酸的含量

目的测定陕西境内不同产地、不同生长期女贞子中齐墩果酸与熊果酸的含量。方法采收陕西境内关中3市(渭南、西安、宝鸡)与陕南2市(安康、汉中)10月产女贞子;并采收西安8、9、10、11和12月产女贞子,去杂质阴干,室温密闭贮藏。采用HPLC法,Lichrospher C_(18)(4.6 mm×250 mm,5μm)色谱柱,Waters600高效液相色谱仪,流动相为乙腈-甲醇-水-磷酸-三乙胺(50:30:20:0.02:0.04),流速1mL/min。建立标准工作曲线,检测各地、各时间点样品中齐墩果酸和熊果酸含量。结果安康产女贞子中齐墩果酸与熊果酸含量均达到同期最高。不同生长期女贞子在10月份出现齐墩果酸含量最高值;而熊果酸在幼果中(8月)含量最高并随生长期延长逐渐下降。到12月时二者含量均降至最低。结论女贞子在不同产地、不同生长期齐墩果酸和熊果酸的含量是不同的,故在实际应用时应根据不同需要适时采收。     

女贞子墨旱莲对多发性肌炎模型大鼠肌组织中核转录因子-κB的影响

目的研究中药女贞子墨旱莲配伍对多发性肌炎模型大鼠的治疗作用与疗效,阐明养阴滋补肝肾法对多发性肌炎模型大鼠肌组织中核转录因子-κB(NF-κB)的影响。方法将40只SPF级SD雄性大鼠随机抽取10只作为空白组,其余大鼠皮下注射完全弗氏佐剂与抗原1∶1混合乳化液,成功制备多发性肌炎大鼠模型,成模大鼠随机分为中药组、激素组、模型组,各10只。空白组、模型组给予0.9%氯化钠注射液,激素组给予甲泼尼龙溶液,中药组给予女贞子墨旱莲药物溶液,均给药4周,观察大鼠的一般状态,根据Lennon LA的评分标准进行评分,检测血清中血清肌酸激酶(CK)、乳酸脱氢酶(LDH)、天冬氨酸氨基转移酶(AST)含量。采用酶联免疫吸附法(ELISA)检测肌肉组织中NF-κB的含量水平。结果在各组大鼠进行干预治疗后,中药组的症状评分低于模型组(P0.01),中药组AST水平明显低于模型组(P0.01),中药组与激素组比较差异无统计学意义(P0.05)。中药组LDH水平明显低于模型组(P0.01),中药组与激素组比较,血清中LDH含量较高(P0.05)。中药组CK水平明显低于模型组(P0.01),中药组与激素组,血清中CK含量高(P0.05)。中药组的NF-κB表达水平明显低于模型组(P0.05),中药组与激素组比较差异无统计学意义(P0.05)。结论中药女贞子墨旱莲阻止炎症细胞凋亡,进而起到抗炎、阻止肌肉组织破坏,是通过抑制NF-κB信号通路,促进抗凋亡蛋白的表达,从而达到对多发性肌炎的治疗作用,表明养阴滋补肝肾法对多发性肌炎模型大鼠肌组织中NF-κB有一定的影响。     

RP-HPLC determination of water-soluble active constituents and oleanolic acid in the fruits of Ligustrum lucidum Ait. collected from various areas]

The contents of p-hydroxyphenethyl-beta-D-glucoside, specnuezhenide and oleanolic acid in the fruits of Ligustrum lucidum collected from various areas in China were determined by reverse phase high performance liquid chromatography. The results could be used to evaluate the quality of the crude drug more comprehensively.     

Antidiabetic and antioxidant effects of extracts from Potentilla discolor Bunge on diabetic rats induced by high fat diet and streptozotocin

Aim of the study

Potentilla discolor Bunge, commonly found at the north temperate and boreal zone, has been used for diabetes in China for a long time. Flavonoids and triterpenoids are two major types of compounds in P. discolor. This study was designed primarily to investigate the effects of total flavonoids extract (TFE) and total triterpenoids extract (TTE) of P. discolor Bunge on blood glucose, lipid profiles and antioxidant parameters on diabetic rats induced by high fat diet and streptozotocin.

Materials and methods

High fat diet-fed and streptozotocin-induced diabetic rats were treated with the TFE and TTE for 15 days, respectively. A range of parameters were tested including fasting blood glucose (FBG), serum insulin (SI), blood lipid profile, malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), glycosylated serum protein (GSP), and nitric oxide (NO).

Results

Diabetic rats treated with TFE or TTE had decreased concentration of FBG and GSP compared with the control group. Meanwhile, the levels of serum total cholesterol (TC), triglycerides (TG) and low-density lipoprotein cholesterol (LDL-c) in the TFE or TTE treated diabetic rats were lower, and the high-density lipoprotein cholesterol (HDL-c) level was higher than in the control diabetic rats. Furthermore, the extracts treatment decreased the MDA and NO level, while increased SOD and GSH levels in diabetic rats. Histopathologic examination also showed that the extracts have protective effects on β-cells in diabetic rats which are supported by the increase of SI.

Conclusions

All these experimental results highlighted the hypoglycemic and hypolipidemic properties of the two extracts from Potentilla discolor Bunge on diabetes and its complications, possibly through a strong antioxidant activity and a protective action on β-cells.     

番石榴多糖对糖尿病小鼠的血糖及抗氧化能力的影响

目的:观察番石榴多糖对四氧嘧啶诱导的糖尿病小鼠血糖及抗氧化水平的影响。方法:通过对小鼠腹腔注射四氧嘧啶建立糖尿病小鼠模型,将小鼠分成5组:正常对照组、糖尿病对照组、番石榴多糖Ⅰ组、番石榴多糖Ⅱ组、格列苯脲组。在给小鼠灌胃多糖3 d和10 d后,采用血糖仪检测小鼠血糖值,第10日麻醉处死小鼠,分别检测小鼠血清和肝脏中的SOD和MDA水平。结果:与糖尿病对照组比较,灌喂番石榴多糖的两组小鼠的血糖值均较低,同时两组小鼠的血清及肝脏中的T-SOD含量较高,MDA含量较低。结论:番石榴多糖能够显著降低糖尿病小鼠血糖,提高其抗氧化能力,是一种潜在的糖尿病治疗药物。     

HPLC-PAD法测定市售六味地黄丸中熊果酸和齐墩果酸的含量

目的:通过测定不同厂家六味地黄丸(熟地黄、山药、山莱萸、泽泻、等)中熊果酸、齐墩果酸的含量,探讨二者对控制该药质量的影响。方法:取市售不同厂家的六味地黄丸,采用超声提取法提取试样,以HPLC-PAD法测定其产品中熊果酸、齐墩果酸含量。色谱柱为Kromasil C18(4.6mm×250mm,5μm),流动相:甲醇-水-磷酸(88:12:0.02),流速:1mL/min。结果:熊果酸进样量在6.462～0.718μg,齐墩果酸进样量在3.132～0.348μg时,呈良好的线性关系,平均加样回收率分别为99.73和97.59,RSD分别为1.5和1.7;4个厂家产品其熊果酸、齐墩果酸含量之间存在显著差异。结论:本试验建立的方法简便,快速,可行。不同厂家的六味地黄丸产品,熊果酸、齐墩果酸含量存在显著差异,但熊果酸、齐墩果酸两者的含量,其高低呈一致性。     

齐墩果酸对大鼠体内细胞色素P450酶活性的影响

目的：研究齐墩果酸对大鼠体内细胞色素P450酶CYP3A4的影响。方法：用Cocktail探针药物法,将SD大鼠按体重随机分组,分别给予0．5％CMC—Na、齐墩果酸高剂组、中剂组和低剂组灌胃9天,第10天给药1h后,灌胃给药维拉帕米,不同时间点采集血样,用LC—MS／MS法测定血浆中探针药物维拉帕米及其代谢产物去甲维拉帕米的浓度,用PKSolution软件估算其药代动力学参数。结果：连续灌胃给药齐墩果酸10天,维拉帕米和去甲维拉帕米药动学参数Cmax、AUC显著减小（P〈0．01）。结论：连续灌胃给药齐墩果酸10天,对CYP3A4酶活性有显著诱导作用。     

HPLC法测定白芪龙胶囊中齐墩果酸和熊果酸

目的 建立白芪龙胶囊中齐墩果酸和熊果酸的定量测定方法.方法 色谱柱Kromasil C18柱(4.6 mm×250 mm,5μm);流动相甲醇-乙腈-0.1％冰醋酸(80∶5∶15);检测波长215 nm;体积流量1.0 mL/min;柱温40℃.结果 齐墩果酸和熊果酸分别在0.410 4～3.078 0μg(r=0.999 9)、0.814 4～6.108 0 μg(r=0.999 9)范围内线性关系良好,回归方程分别为Y=490 276.942 1X-1 265.765 3和Y=457 259.686 7X+14 380.9603,平均加样回收率分别为98.60％(RSD为1.35％)和99.40％(RSD为1.19％).结论 所建立的方法准确可靠,重复性好,可用于白芪龙胶囊的质量控制.