Ameliorative effects of standardized extract from Trigonella foenum-graecum L.seeds on painful peripheral neuropathy in rats

Objective:To evaluate the effects of the standardized extract of fenugreek(Trigonella foenumgraecum L.Family:Leguminasae) seed(IND01) in animal models of peripheral neuropathy. Methods:IND01 was prepared from fenugreek seeds and standardized by high performance liquid chromatography to a marker compound,trigonelline.The effects of daily oral administration of IND01(50,100 and 200 mg/kg) were studied in rats after partial sciatic nerve ligation(PSNL) and sciatic nerve crush injury(SNCI) during 30-days period.The measurements on thermal hyperalgesia(TH),motor function test(MFT) score and motor nerve conduction velocity (MNCV) were recorded.Results:IND01 offered sustained protection against TH and deranged MFT scores in both models from 7-day onwards.Fifteen days of daily oral administration of IND01 restored MNCV reduction in rats with SNCI but not with PSNL.Conclusions:IND01 was found to be effective in rat models of painful peripheral neuropathy.

Aqueous and organic extracts of Trigonella foenum-graecum L.inhibit the mycelia growth of fungi

Aqueous extracts from various plant parts of fenugreek(3%)(aerial parts:leaves and stems(LS),roots(R),ground seeds(GS)and not ground seeds(NGS))and petroleum ether,ethyl acetate and methanolic fractions of the aerial parts were assayed to determine their antifungal potential against Botrytis cinerea,Fusarium graminearum,Alternaria sp.,Pythium aphanidermatum,and Rhizoctinia solani.All fenugreek plant parts showed antifungal potential and the magnitude of their inhibitory effects was species and plant parts d...

Distribution, Enrichment and Accumulation of Heavy Metals in Soil and <i>Trigonella foenum-graecum</i>L. (Fenugreek) after Fertigation with Paper Mill Effluent

The aim of the study was to investigate distribution, enrichment and accumulation of heavy metals in soil and Trigonella foenum-graecum (var. Pusa Early Bunching) after fertigation with paper mill effluent. Doses of paper mill effluent viz. 5%, 10%, 25%, 50%, 75% and 100% were used for fertigation of T. foenum-graecum along with bore well water (control). The results revealed that paper mill effluent had significant (P +, K+, Ca2+, Mg2+, Fe2+, TKN, PO3-4, SO2-4, Cd, Cr, Cu, Mn and Zn of the soil in both seasons. Insignificant (P > 0.05) changes in WHC and bulk density of the soil were observed after irrigation with paper mill effluent. The agronomical performance of T. foenum-graecum was increased from 5% to 25% concentration and decreased from 50% to 100% concentration of paper mill effluent as compared to control in both seasons. The heavy metals concentration was increased in T. foenum-graecum from 5% to 100% concentrations of paper mill effluent in both seasons. Biochemical components like crude proteins, crude fiber and crude carbohydrates were found maximum with 25% paper mill effluent in both seasons. The enrichment factor (Ef) of various heavy metals was in order of Cd > Mn > Cr > Cu > Zn > Fe for soil and Mn > Cu > Cr > Cd > Zn > Fe for T. foenum-graecum plants after fertigation with paper mill effluent. Therefore, paper mill effluent can be used as a biofertigant after appropriate dilution to improve yield of T. foenum-graecum.

The impact of cold atmospheric pressure plasma jet on seed germination and seedlings growth of fenugreek(Trigonella foenum-graecum)

The effects of cold atmospheric-pressure plasma jet (CAPPJ) were investigated on germination and early seedling growth of fenugreek (Trigonella foenum-graecum L) seeds. A two-electrode argon CAPPJ system with and without an additional grounded electrode [accelerating grounded (AG) electrode] was used at different exposure times. After 16 h of observation, the germination rates increased by 4 and 7 times, without and with using an AG electrode, respectively, for 1 min of plasma exposure. An increase in shoot fresh weight was observed, especially at 10 min exposure time. A high dry weight of root and shoot at 1 min-AG exposure time was observed. The root:shoot ratio was lower in plasma-treated seedlings, compared with the control plants. The study found that the O-radical emission line (777.4 nm) enhanced 5 times, due to the presence of an AG electrode, which increased the axial electric field and led to the formation of more streamers. The three stated effects (O-radicals, enhancement of the electric field and streamers) could be the cause for the stimulation of seed germination and seedling growth parameters when using the CAPPJ. The scanning electron microscope images showed the etching of the seed surface layers, which was more pronounced when an AG electrode was applied. The results of the current study indicate that the germination rates increased due to the increase of O-radical concentration and the etching of the seed surfaces.

Ameliorative properties of Iranian Trigonella foenum-graecum L. seeds and Punica granatum L. peel extracts in streptozotocin-induced experimental diabetic guinea pigs

Objective: To assess the ameliorative properties of Iranian Trigonella foenum-graecum L.(T. foenum-graecum) seeds and Punica granatum L.(P. granatum) peel extracts against streptozotocin-induced diabetes in male guinea pigs.Methods: Sixty guinea pigs were divided into six groups(10 guinea pigs per group).Group 1 consisted of normal animals. Groups 2 and 3 were treated with Iranian T. foenum-graecum seeds and P. granatum peel extract alone, respectively. Group 4 was treated with streptozotocin only; whereas Groups 5 and 6 receiving streptozotocin were treated with Iranian T. foenum-graecum seeds and P. granatum peel extract, respectively.All animals were treated for 30 days, and the body weight, blood and liver biochemical parameters were measured.Results: Guinea pigs exposed to streptozotocin showed an alteration in body weight gain, fasting glucose level, kidney function parameters(blood urea nitrogen and creatinine) as well as decreased serum and hepatic total protein level. In addition, it increased the cholesterol and triglyceride level, while decreasing the hepatic glucose-6 phosphate dehydrogenase activity, glycogen, glutathione content and hepatic catalase activity. Oral treatment with T. foenum-graecum seeds and P. granatum peel extracts revealed significant protective properties with respect to body weight gain and other biochemical parameters studied.Conclusions: The Iranian T. foenum-graecum seeds and P. granatum peel extracts are significantly potent in ameliorating diabetic condition induced by streptozotocin and improving various biochemical parameters in serum and liver of guinea pigs.

NaCl胁迫对胡卢巴幼苗抗氧化酶活性和丙二醛含量的影响

以胡卢巴幼苗为试验材料,研究NaCl胁迫下胡卢巴幼苗叶和根中抗氧化酶SOD、POD、CAT活性及丙二醛(MDA)含量的变化。结果表明:随着NaCl处理浓度的升高,SOD、POD、CAT活性总体呈现先上升后下降的趋势,MDA含量则呈上升趋势。叶片中抗氧化酶活性在NaCl浓度为1.0%时达到峰值,根中抗氧化酶活性在NaCl浓度为0.5%时达到峰值。在0.5%的NaCl浓度处理下,随着处理时间的延长,3种抗氧化酶协同作用,使MDA含量减少并控制在较稳定的阶段。可见,盐胁迫下胡卢幼苗可通过提高抗氧化酶活性,降低膜质过氧化水平,减缓盐胁迫对植株的伤害,从而增强其耐盐性。

葫芦巴4-羟基异亮氨酸对高糖诱导小鼠3T3-L1脂肪细胞胰岛素抵抗的影响

目的研究葫芦巴活性成分4-羟基异亮氨酸(4-hydroxyisoleucine,4-HIL)对高糖诱导的小鼠前脂肪细胞(3T3-L1)胰岛素抵抗的作用及其机制。方法以25 mmol/L葡萄糖加0.6 nmol/L胰岛素诱导小鼠3T3-L1脂肪细胞建立胰岛素抵抗细胞模型,予以不同浓度4-HIL(5、10、20μmol/L)进行干预。采用[3H]-脱氧-D-葡萄糖摄入法检测葡萄糖摄取率,PCR法检测肿瘤坏死因子-α(tumor necrosis factor-alpha,TNF-α)mRNA表达量,ELISA法检测细胞上清TNF-α蛋白水平。以棕榈酸(palmitic acid,PA)为对照。结果 25mmol/L葡萄糖加0.6 nmol/L胰岛素作用18 h后,3T3-L1脂肪细胞胰岛素刺激的葡萄糖转运抑制了63%;PCR显示脂肪细胞内TNF-αmRNA的表达较正常脂肪细胞明显增高(P<0.05);ELISA显示细胞上清中的TNF-α分泌量增加70 pg/mL(P<0.05)。PA组表现出类似结果。分别加入5、10、20μmol/L 4-HIL作用24h后,3T3-L1脂肪细胞胰岛素刺激所产生的葡萄糖转运分别增加35%、50%、60%;PCR检测显示,脂肪细胞内TNF-αmRNA的表达随着药物浓度的增加呈递减趋势,与模型组及PA组比较,差异有统计学意义(P<0.05);与模型组比较,药物干预后脂肪细胞培养基上清TNF-α分泌水平分别下降10、18、39 pg/mL(P<0.05)。结论 4-HIL可以显著改善高糖诱导的小鼠3T3-L1脂肪细胞胰岛素抵抗,同时4-HIL能够抑制TNF-α的分泌。

胡芦巴药材HPLC指纹图谱的初步研究

目的研究胡芦巴药材的高效液相指纹图谱,为科学评价及有效控制药材的质量提供了可靠的方法。方法利用高效液相色谱法,梯度洗脱,测定了14个不同产地的胡芦巴药材样品。色谱条件为:Alltech C18柱(4.6 mm×250 mm,5μm),流动相:乙腈-水,流速:1 mL.min-1,检测波长:211 nm。结果各产地胡芦巴药材指纹图谱有21个共有峰,峰面积之和大于总峰面积的90%,经相似度计算,各产地药材之间的相似性良好。结论方法灵敏度高,重现性好,可作为控制胡芦巴药材内在质量的标准。

NaCl胁迫对胡卢巴幼苗光合生理特性的影响

以胡卢巴幼苗为试验材料,分别选用浓度0、0.2%、0.5%和1.0%NaCl的Hoagland营养液对胡卢巴幼苗进行盐胁迫处理,探讨不同浓度NaCl胁迫对胡卢巴幼苗光合生理特性的影响。结果表明,随着NaCl浓度增加,胡卢巴幼苗叶片净光合速率(Pn)、蒸腾速率(Tr)和气孔导度(Gs)均呈降低趋势;胞间CO(2Ci)浓度先降低,随后在1.0%NaCl浓度胁迫下有所增加;叶绿素含量呈先升高后降低的趋势,在0.5%NaCl浓度胁迫下达到最大值;而叶绿素a/b比值则一直呈增加趋势。叶片细胞膜伤害度在0.2%NaCl浓度胁迫下较低,当NaCl浓度升至0.5%时显著增加,但在1.0%NaCl浓度胁迫下仍维持在小于8.0%的水平。

HPLC法测定葫芦巴中薯蓣皂苷元的含量

建立葫芦巴中薯蓣皂苷元含量的测定方法。采用高效液相色谱法,色谱柱为Agilent Technologies 1200 Series C18柱（4.6mm×150mm,5μm）,流动相为乙腈,柱温30℃,流速1.0mL·min^-1,检测波长209nm。葫芦巴薯蓣皂苷元在9.00～54.00μg·mL^-1范围内峰面积与其浓度线性关系良好（r=0.9952）,平均加样回收率为108.4%,精密度RSD为1.21%（n=5）。该方法快速、准确、可靠,为葫芦巴药材质量控制提供了定性和定量依据。

柱前衍生化HPLC法测定不同产地胡芦巴中4-羟基异亮氨酸的含量

目的柱前衍生化法测定4-羟基异亮氨酸含量。方法采用异硫氰酸苯酯作为衍生化试剂,色谱柱(Zorbax Eclipse XDBC18,150mm×4.6mm,5μm,Agile);流动相A:乙腈,流动相B:1mL·L^(-1)磷酸水,梯度洗脱(A相∶B相):(0~20min,20∶80;20min,60∶40);流速:1.5mL·min^(-1);进样体积:20μL;检测波长:254nm;柱温:30℃。结果 4-羟基异亮氨酸在质量浓度4.2~20.9μg·mL^(-1)范围内线性关系良好(r=0.999 3),平均回收率为96.8%,RSD为1.7%。结论该方法简便、稳定,结果准确,可作为4-羟基异亮氨酸的含量测定方法。

UPLC-QTOF-MS 结合主成分分析法考察胡芦巴盐制前后的化学成分差异

目的 利用超高效液相-四极杆-飞行时间串联质谱(UPLC-QTOF-MS)结合多元统计分析筛选并鉴定胡芦巴和盐胡芦巴的差异性成分。方法 采用C_(18)反相色谱柱0.5%乙酸-乙腈梯度洗脱,在正离子模式下采集质谱数据,应用Simca-P等软件进行主成分分析(PCA)和偏最小二乘判别分析(PLS-DA),筛选胡芦巴与盐胡芦巴的差异性成分。通过质谱提供的精确分子量、碎片离子并结合文献数据鉴定偏最小二乘判别分析中VIP值>1.5的差异性成分的结构。结果 利用准分子离子、二级质谱图、特征碎片离子及其他碎片离子等信息,并参考相关文献报道鉴定了胡芦巴与盐胡芦巴中的24个差异性成分,其中包括7个黄酮碳苷类化合物,15个甾体皂苷类化合物以及1个生物碱类化合物即胡芦巴碱和1个脂肪族化合物即亚麻酸乙酯。通过比较各化合物在胡芦巴和盐胡芦巴样品的质谱中的响应值,盐制后黄酮碳苷类成分响应升高,甾体皂苷中的呋甾烷醇型甾体皂苷响应降低,而螺甾烷醇型/异螺甾烷醇型甾体皂苷响应升高。结论 盐制促进了胡芦巴中黄酮碳苷类成分的溶出,并使胡芦巴中皂苷类成分发生了结构转化。

葫芦巴(Trigonella foenum-graecum)染色体核型分析

葫芦巴是现在较为常用的中药材之一,但对其染色体的核型分析研究较少。为了研究葫芦巴染色体形态结构特征以及探索遗传机制,本试验以采自河北的葫芦巴种子为研究对象,采用根尖染色体制片与显微摄影技术相结合的方法,以确定染色体数目和进行核型分析。结果显示:河北葫芦巴有16条染色体,核型公式为2n=16=4m+2m (SAT)+10sm,染色体相对长度在4.63%~8.17%范围内,臂比在1.17~2.43范围内,核型不对称系数为64.03%,随体在第8对染色体的短臂上,核型类别为2A型。本研究结果既可为今后选择种内杂交和种间杂交育种种类提供科学依据,又可为探索其物种遗传机制、亲缘关系与进化、远缘杂种的鉴定等提供重要依据。

胡芦巴油脂成分的分析测定

应用ＧＣ－ＭＳ技术对国产胡芦巴油脂成分进行了分析研究，鉴定了５种脂肪酸甲酯和１７种油脂类成分，其中１５种成分为首次从胡芦巴中测得。分析结果为开发利用胡芦巴油脂资源提供了依据。

亲水作用色谱法测定胡芦巴中的胡芦巴碱

建立了亲水作用色谱法(HILIC)测定胡芦巴药材中胡芦巴碱含量的方法。采用Waters Atlantis HILICS ilica色谱柱(150mm×2.1mm,3μm),以乙腈-乙酸铵溶液(pH4.4)(体积比为70:30)为流动相,流速0.4mL/min,检测波长265nm。胡芦巴碱的线性范围为2.50～100mg/L(r=0.9996);两个加标水平的平均加样回收率为102%,相对标准偏差(RSD)分别为4.17%和2.28%(n=3)。结果表明所建方法分离效果好、快速简易,可以弥补中国药典中离子对色谱法(IPLC)平衡时间过长的缺陷,适用于胡芦巴药材中强极性胡芦巴碱的测定,为胡芦巴的质量控制提供了有效的方法。

胡芦巴种子化学成分的研究

对胡芦巴 (Trigonellafoenum graecumL .)种子的物理组成及主要化学组成进行了研究 .实验结果表明 ,胡芦巴种子物理组成中 ,种皮占 15 5 2 %,内胚乳占 31 14 %,胚芽、子叶占 5 3 34%;种子千粒重为 14 78%,容重为2 0 0 7%.测定了胡芦巴种子 18种氨基酸和 8种微量金属元素的含量 .粗蛋白含量为 5 4 0 6 %,种子中聚糖含量为8 72 %.

胡芦巴不同溶剂粗提物对两种储粮害虫的驱避和触杀活性

为开发环境和谐的植物性杀虫剂,以乙醇、乙酸乙酯、丙酮、氯仿、石油醚为溶剂采用温浸回流法对胡芦巴Trigonella foenum-graecumL.种子中的生物活性物质进行提取;并以主要储粮害虫杂拟谷盗Triboli-um confusumdu Val、谷蠹Rhyzopertha dominicaFbricius成虫为试验对象,采用滤纸药膜法对胡芦巴5种溶剂粗提物进行驱避、触杀活性测定。结果表明:5种溶剂粗提物对杂拟谷盗的驱避作用均优于对谷蠹的作用:在各粗提物剂量为1.6μg/cm2时,对杂拟谷盗处理7 d后的平均驱避率均超过80%,驱避等级为Ⅴ级,其中丙酮粗提物效果最好。但5种溶剂粗提物对谷蠹的触杀作用却均优于对杂拟谷盗的作用:处理72 h后,乙醇、乙酸乙酯、丙酮、氯仿粗提物对谷蠹成虫的LD50依次为0.112、0.102、0.106、0.100 mg/cm2,石油醚粗提物的触杀活性相对较差;乙醇、乙酸乙酯、氯仿粗提物用0.91 mg/cm2剂量处理72 h后,杂拟谷盗成虫的平均死亡率分别为95.56%、72.22%、87.78%;丙酮、石油醚粗提物用1.04 mg/cm2剂量处理72 h后,杂拟谷盗成虫的平均死亡率分别为57.78%和53.33%。

酶解法提取胡芦巴种子中薯蓣皂苷元的工艺研究

研究了酶解结合水浸提法从脱脂胡芦巴种子中提取薯蓣皂苷元的工艺。以薯蓣皂苷元的提取率为指标,优化了该法制备薯蓣皂苷元的工艺条件,得到以下结论:与水浸提法相比,采用酶解法有利于提高皂苷元的提取率;确定料液比为1∶14,纤维素酶(每克原料10U,pH值4.6,45℃)酶解6h为最佳工艺条件。在此基础上考察了酶解结合水浸提法对皂苷元提取率的影响,发现酶解后升温至90～100℃,保温水浸提70min,可使薯蓣皂苷元的提取率由83.8%提高到92.9%。

发酵对葫芦巴挥发性化学成分的影响

采用水蒸气蒸馏与溶剂萃取相结合的方法对未发酵和发酵后的葫芦巴茎叶挥发性成分进行提取,运用毛细管气相色谱-质谱联用法分析测定,用计算机检索NIST02质谱数据库确定其挥发性化学成分,用面积归一化法进行定量分析。结果表明,两种方法提取的挥发性化学成分及含量皆有很大差异,发酵的葫芦巴挥发油产率明显提高。经毛细管色谱从未发酵的葫芦巴茎叶挥发油中分离出93个峰,鉴定出65种成分,占挥发油总相对含量的69.89%,从发酵后的葫芦巴茎叶挥发油中分离出134个峰,鉴定出49种成分,占挥发油总相对含量的36.57%,二者共有成分32种,这与已有文献结果存在明显的区别。

响应面法优化闪式提取葫芦巴多糖及其保润性能研究

利用闪式提取方法,以葫芦巴多糖（FPS）提取率为响应值,首先通过单因素实验考察液料比、提取时间、温度、提取电压和提取次数的影响,初步优化提取条件;进而,通过Box-Behnken实验设计和响应面分析法综合优化最终提取条件;并对葫芦巴多糖进行卷烟物理和感官保润性能测试。结果表明,葫芦巴多糖的最佳提取条件为：液料比27∶1（m L/g）,闪式提取时间136 s,提取温度58℃,电压140 V;在最优条件下进行了5次验证实验,葫芦巴多糖的平均提取率为21.23%,与理论值（21.80%）的相对误差为2.61%,响应面优化闪式提取葫芦巴多糖的方法可行;葫芦巴多糖作为保润剂在相对湿度较低的环境下,对烟丝的保湿能力强于丙二醇样品,在湿度较高的环境下,葫芦巴多糖具有防潮效果;且能够改善卷烟吸食品质,葫芦巴多糖适宜的添加量为烟丝质量的0.005%~0.1%。