几种药用芳香植物的抗氧化、抗菌、降糖和抗癌活性

发布时间：2018-02-04 11:30

本文关键词： Pulicaria iuloides 精油抗氧化抗菌抗糖尿病抗癌　出处：《江南大学》2017年博士论文　论文类型：学位论文

【摘要】：Pulicaria inuloides是Asteraceae科Pulicaria属植物,它被广泛用作抗癌药物,并用于治疗糖尿病、风湿、痛风、膀胱炎、肾炎、痛经、牙痛、头痛等,并具有抗菌,抗凝血等功效。P.inuloides含有生物碱、挥发油、萜类、酚酸、类黄酮、单宁酸等成分。第一阶段,从药用芳香植物Pulicaria inuloides-Astereceae和Ocimum forskoleiLamaiceae叶子中得到精油,从精油中分离并鉴定生物活性物质。然后测定分离出的物质的抗氧化活性、总酚含量和抗微生物活性。结果表明与O.forskolei相比,P.inuloides有较高的总酚量(O.forskolei为54.6±30 mg GAE/g提取物,P.inuloides为144±5.32 mg GAE/g提取物);P.inuloides具有较高的抗氧化活性,DPPH清除率分别为26.76±0.11%(O.forskolei)和92.92±0.10%(P.inuloides),IC50分别为73.03±0.05(O.forskolei)和4.5±0.05(P.inuloides);P.inuloides有更高的β胡萝卜素漂白值:分别为41.03±6.35%(O.forskolei)和90.77±0.21%(P.inuloides)。P.inuloides的精油能够抑制除鼠伤寒沙门氏菌和志贺氏痢疾杆菌以外的所有测试微生物,并且对大肠杆菌的最小抑制浓度(MIC)为3.0μg/m L。O.forskolei精油仅抑制白色念珠菌。第二阶段,通过用气相色谱-质谱联用法鉴定P.inuloides和O.forskolei根中的挥发性化合物结果表明,与O.forskolei相比,P.inuloides根中含有较高的总酚含量(P.inuloides为55.4±0.1,O.forskolei为35.3±0.2mg GAE/g提取物);P.inuloides具有较低的抗氧化活性,DPPH清除率分别为23.07±0.06%(P.inuloides)43.97±0.18%(O.forskolei),IC50值分别为80.01±0.03(P.inuloides)和31.18±0.01(O.forskolei);P.inuloides有较低的β胡萝卜素漂白值:分别25.3±0.3%(P.inuloides)和39.9±0.7%(O.forskolei)。P.inuloides精油能抑制除鼠伤寒沙门氏菌,痢疾杆菌和大肠杆菌以外的所有测试微生物。在第三阶段,研究溶剂类型对Pulicaria inuloides(Arecaceae)中多酚、单宁、花青素和类黄酮提取的影响,以及对其抗氧化和抗菌活性影响的研究。提取中分别采用超声处理30分钟,频率为50 Hz,速度为70 rpm。结果表明,总多酚含量(TPC)、总黄酮(TFC)和总的丹宁含量(TTC)以及抗氧化能力在使用甲醇提取时达到最大。P.inuloides的甲醇和乙醇提取物对于所有选定的微生物都具有较低的抗菌活性。二乙醚提取物对选定的所有被测试细菌没有抗菌活性。第四阶段,蚤草属inuloides叶的乙醇提取物的GC-MS分析显示10峰指示存在10个化合物。第五阶段,研究从P.inuloides中提取分离的酚类化合物的抗氧化和抗菌活性,并对其脂肪酸和生育酚含量进行评价。通过大孔树脂柱层析分离得到酚类化合物,然后利用高效液相色谱法进行分析。提取物中γ-生育酚具有最高含量,大约占总生育酚的52.76%,其次是α-生育酚(29.83%)和δ-生育酚(17.41%)。另外,P.inuloides中含有大量的钾,镁,钠,铁和钙,含量分别为159.5,29.5,14.2,13.875和5.225mg/100g。利用扫描电镜、透射电镜(TEM,SEM and EDX)揭示了P.inuloides的内部结构。第六阶段,将P.inouloides叶子中的精油(PIEO)应用在海产品juvenile kachlan(Trachinotus ovatus,Linnaeus)上。结果表明,P.inuloides的精油抑制所有测试的微生物,对大肠杆菌的最小抑制浓度(MIC)为2.0μg/m L。本研究以纯水为对照组,研究了Pi EO应用在鱼片上对单核细胞增生李斯特氏菌、大肠杆菌和金黄色葡萄球菌的影响。结果表明,以纯水做参照,应用了Pi EO后(25℃),立即消除了鱼片上大部分单核细胞增生李斯特氏菌、大肠杆菌和金黄色葡萄球菌,降低了固相细胞计数值(SPC)。在10℃下12天内也可减少以上细菌的含量,在10℃保存条件下,延长了食品的货架期。在贮藏结束时,SPC计数7 log cfu/g。感官评价证明P.inouloides精油处理后的鱼片是可以被消费者接受的。结果还表明,从超市(江苏,无锡)购买的鱼产品拥有良好的卫生及健康条件,而从当地市场购买的鱼产品则不达标准。在下一阶段,验证P.inouloides叶中的精油对于糖尿病的效果。精油的化学分析结果表明它的成分中含有生物碱、类固醇、皂甙、糖苷、还原糖和萜类化合物。在250μg/ml的浓度下,P.inuloides叶子精油对α-淀粉酶和α-葡糖苷的抑制活性分别为85.50%和86.52%,IC50值分别为140.49μg/ml和128.33μg/ml。P.inuloides叶子精油在实验剂量水平(400 mg/kg体重)下可以降低糖尿病大鼠的空腹血糖水平。到了实验的第21天,在P.inuloides叶子精油处理组,大鼠血糖值下降到与糖尿病药物处理组大鼠没有明显差异(P0.05)。在链脲佐菌素(STZ)诱导糖尿病模型大鼠中,P.inuloides叶子精油可以降低大鼠的总胆固醇,甘油三酯和低密度脂蛋白(LDL)水平,同时会增加高密度脂蛋(HDL)水平。在实验的21天内,大鼠体重保持稳定。在链脲佐菌素(STZ)诱导糖尿病模型大鼠中,反应肝功能的标志物谷丙转氨酶(ALT)、谷草转氨酶(AST)和碱性磷酸酶(ALP)含量的显著升高。而P.inuloides叶子精油的处理可以显著降低大鼠体内这些酶的水平(P0.01)。在链脲佐菌素(STZ)诱导糖尿病模型大鼠中,肌酐、尿素氮、总蛋白等肾功能标记物含量显著升高。而P.inuloides叶子精油的处理可以显著降低大鼠体内以上指标的水平(P0.01)。这些结果表明,糖尿病大鼠都患有肾功能障碍。肝脏组织病理学研究表明,与对照组相比,P.inuloides叶子精油能广泛改善胰岛细胞的生理状况。P.inuloides的毒理试验结果显示,P.inuloides没有任何导致急性毒性的迹象,如呼吸窘迫,昏迷或死亡等。在最后一部分细胞实验中,结果显示对P.inuloides叶子精油MCF-7细胞,具有很强的细胞毒活性的作用,IC50分别为12.9和3.00μl/ml。Pi EO的精油抑制所有测试的微生物(Listeria)。并运用光学显微镜对MCF-7细胞形态进行了分析。
[Abstract]:Pulicaria inuloides Asteraceae is a plant of the genus Pulicaria, which is widely used as anticancer drugs, and for the treatment of diabetes, rheumatism, gout, cystitis, nephritis, dysmenorrhea, toothache, headache, and has antibacterial, anti coagulation effect of.P.inuloides containing alkaloids, terpenes, volatile oil, flavonoids, phenolic acid, tannic acid and other ingredients. Stage, get the essential oil from Pulicaria inuloides-Astereceae and Ocimum of medicinal and aromatic plants leaves forskoleiLamaiceae, isolation and identification of bioactive substances from oil. Then the determination of antioxidant activity of the isolated material, and the anti microbial activity of total phenolic content. The results showed that compared with O.forskolei, P.inuloides had higher total phenolic content (O.forskolei 54.6 + 30 mg GAE/g extract, P.inuloides 144 + 5.32 mg GAE/g extract); P.inuloides has the higher antioxidant activity, DPPH clearance rates were 26.76 + 0.11% (O.forskolei) and 92.92 + 0.10% (P.inuloides), IC50 were 73.03 + 0.05 and 4.5 + 0.05 (O.forskolei) (P.inuloides); P.inuloides has a higher beta carotene bleaching values were 41.03 + 6.35% and 90.77 + 0.21% (O.forskolei) (P.inuloides).P.inuloides essential oil can inhibit all the tested microorganisms except Salmonella and Shigella dysenteriae, and the minimum inhibitory concentration of Escherichia coli (MIC) 3 g/m L.O.forskolei oil only inhibit Candida albicans. The second stage, by using gas chromatography - mass spectrometry method for identification of P.inuloides and volatile compounds in the roots of O.forskolei. The results showed that compared with O.forskolei, total phenol the content of P.inuloides in root contains higher (P.inuloides = 55.4 + 0.1, 35.3 + O.forskolei 0.2mg GAE/g extract); P.inuloides has lower antioxidant activity, DPPH clearance rates were 23. 07 + 0.06% (P.inuloides) 43.97 + 0.18% (O.forskolei), IC50 = 80.01 + 0.03 (P.inuloides + 0.01) and 31.18 (O.forskolei); P.inuloides had lower beta carotene bleaching value: 25.3 + 0.3% (P.inuloides + 0.7%) and 39.9 (O.forskolei).P.inuloides essential oil can inhibit the removal of Salmonella typhimurium all the tested microorganisms except bacteria, Bacillus dysenteriae and Escherichia coli. In the third stage, Pulicaria inuloides of the type of solvents (Arecaceae) in polyphenols, tannins, anthocyanins and flavonoids extraction effect, and the effect of antioxidant and antibacterial activity research. Extraction were used in ultrasonic treatment for 30 minutes, frequency is 50 Hz. The speed of 70 rpm.. The results showed that the total polyphenol content (TPC), total flavonoids (TFC) and total tannins (TTC) and antioxidant capacity reached the maximum.P.inuloides of methanol and ethanol extracts of methanol extraction in use For all selected microorganisms with antimicrobial activity is low. Two selected ether extracts of all tested bacteria have antibacterial activity. The fourth stage, the ethanol extract of leaves of the pulicaria inuloides GC-MS analysis showed 10 peaks indicating the presence of 10 compounds. The fifth stage extraction of phenolic compounds isolated from antioxidant P.inuloides and the antibacterial activity, and to evaluate the fatty acid and tocopherol content. Phenolic compounds by macroporous resin column chromatography, high performance liquid chromatography and then use analysis. In the extract of gamma tocopherol has the highest content of total tocopherol 52.76%, followed by alpha tocopherol (29.83%) and delta tocopherol (17.41%). In addition, contains a lot of potassium, P.inuloides sodium, magnesium, iron and calcium, the content of 159.5,29.5,14.2,13.875 and 5.225mg/100g. respectively by scanning electron microscopy, transmission electron microscopy (TEM SEM, and EDX) to reveal the internal structure of P.inuloides. In the sixth stage, the essential oils will the leaves of P.inouloides (PIEO) used in juvenile kachlan (Trachinotus ovatus, seafood Linnaeus). The results show that the P.inuloides essential oils on all tested microorganisms, the minimum inhibitory concentration of Escherichia coli (MIC) 2 g/m L. in this study, using pure water as the control group, the Pi EO application in the study of fish Listeria monocytogenes List S effect of bacteria, Escherichia coli and Staphylococcus aureus. The results showed that using pure water as the reference, the application of Pi EO (25 C), immediately eliminates most fish Listeria monocytogenes bacteria List S, Escherichia coli and Staphylococcus aureus, reduce solid cell count (SPC). The content of 10 C within 12 days can also reduce bacteria, 10 degrees in the preservation conditions, prolong the shelf life of food in the storage node. Beam, SPC count 7 log cfu/g. P.inouloides after the treatment of the sensory evaluation of fish oil can be accepted by consumers. The results also show that from the supermarket (Jiangsu, Wuxi) has a good health and health conditions to buy fish products, and purchased from the local market of fish products is not up to the standard in the next. Stage, verification of P.inouloides in the leaf essential oil for diabetes. The results showed that the chemical analysis of essential oil composition containing alkaloids, steroids, saponins, glycosides, reducing sugar and terpenoids. At a concentration of 250 g/ml, inhibited P.inuloides activity of leaf essential oils of alpha amylase and alpha glucoside respectively. 85.50% and 86.52%, IC50 values were 140.49 g/ml and 128.33 g/ml.P.inuloides in leaf oil level of experimental dose (400 mg/kg body weight) can reduce fasting blood glucose levels in diabetic rats. On the twenty-first day of the experiment, In the P.inuloides leaf oil treatment group, blood glucose of rats and decreased to the diabetes drug treatment group (P0.05). There was no significant difference in streptozotocin (STZ) induced diabetic rats, P.inuloides leaf essential oil can reduce the total cholesterol of rats, three glycerol ester and low density lipoprotein (LDL at the same time will increase the level), high density lipoprotein (HDL) levels. On the 21 day of the experiment, the body weight of rats remained stable. In streptozotocin (STZ) induced diabetic rats in response, liver function markers alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) significantly increased the content of P.inuloides. And the processing of leaf essential oils can significantly reduce the levels of these enzymes in rats (P0.01). In streptozotocin (STZ) induced diabetic model rats, serum creatinine, urea nitrogen, total protein and other markers of renal function were significantly increased and P.inu. Loides leaf oil can significantly reduce the rats above index level (P0.01). These results showed that diabetic rats had renal dysfunction. The histopathological study showed that, compared with the control group, the toxicological test results of P.inuloides leaf oil can widely improve islet cell physiological status of.P.inuloides showed no P.inuloides any signs of acute toxicity of lead, such as respiratory distress, coma or death. In the last part of the cells in the experiment, the results showed that for P.inuloides leaves oil MCF-7 cells, has strong cytotoxic activity and IC50 were 12.9 and 3 l/ml.Pi EO oils inhibit all the tested microorganisms (Listeria). The use of optical microscope to analyze MCF-7 cell morphology.

【学位授予单位】：江南大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：TQ460.1

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