Protective effects of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja on STZ-stimulated INS-1 cells through regulating of autophagy and apoptosis

OBJECTIVE To explore the protective effects of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja on STZ-stimulated INS-1 cells through regulating of autophagy and apoptosis.METHODS INS-1cells were cultured in media containing 3n M STZ and different doses of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja.The proliferation of cells was examined by MTT assay,ROS content were detected by fluorescence enzyme label.The levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),hydrogen peroxidase(CAT),malondialdehyde(MDA)were also measured by colorimetry.The activities of caspase-3,9 were also observed by Caspase colorimetric assay kit in each group.The expressions of Bcl-2 m RNA and Bax m RNA were detected by Real time PCR,protein expression of LC3-Ⅱand PARP were detected by Western blotting.RESULTS Compared with the control group,total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja could promote the proliferation of INS-1 cells no matter with STZ or not when its concentration lower than 25μg·m L-1;but when its concentration higher than 100μg·m L-(1use individually)or 50μg·m L-1(combined use),total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja might significantly inhibite the growth of the cells whether STZ existed or not.Total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja(6.25,12.5,25μg·m L-1)might inhibit INS-1cell apoptosis,decrease intra-cellular ROS contents,improve the s upernatant liquid SOD,GSH-PX,CAT activities,decrease MDA level,promote INS-1 cell secreting insulin,decrease the protein expressions of autophagy protein LC3Ⅱand apoptosis regulating protein cleaved PARP protein,up-regulate the anti-apoptotic protein Bcl-2 m RNA expression,down-regulate the pro-apoptotic protein Bax m RNA expression and Bcl-2and Bax,reduce the activity of caspase-9 and caspase-3(P<0.05 or P<0.01).CONCLUSION Total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja has good protective effect on STZ-stimulated INS-1cells.It can inhibit STZ injured INS-1 cells to overproduce ROS production,enhance endogenous antioxidant enzymes(GSH-Px,SOD,CAT activities),reduce the expression of autophagy protein LC3Ⅱand apoptosisregulating protein cleaved PARP,up-reglute the antiapoptosis protein Bcl-2 expression,down-regulate the pro-apoptotic protein Bax expression,decrease the caspase-9 and caspase-3 activities,and improve the INS-1cell survival rate,and then play a protective effect on damaged INS-1 cells.

Optimization of Extraction Process and Determination of Total Triterpenoids from Semen Trichosanthis by Response Surface Methodology

[Objectives] To optimize the extraction process and determine the total triterpenoids from Semen Trichosanthis.[Methods]The Box-behnken response surface methodology was applied to optimize the extraction conditions of total triterpenoids in Semen Trichosanthis.And the spectrophotometry was used to determine the content of total triterpenoids in Semen Trichosanthis.[Results] The optimal extraction conditions were solid to liquid ratio of 40∶ 1,ultrasonic time of 20 min,and ultrasonic power of 100 W.Under these conditions,the content of total triterpenoids in Semen Trichosanthis was significantly different among the 10 production areas,among which the No.9 production area(Anhui 1) had the highest total triterpenoids content of 45.71 mg/g,while No.7 production area(Shandong) had the lowest total triterpenoids content of 15.22 mg/g.[Conclusions]The Box-behnken response surface methodology is reliable for the extraction of total triterpenoids in Semen Trichosanthis,and the spectrophotometry is proper for determining the content of total triterpenoids in Semen Trichosanthis.There are large differences in the content of total triterpenoids in Semen Trichosanthis produced in different areas.

Immediate-release Mechanism of Dehydrotrametenolic Acid and Dehydroeburicoic Acid in Poriae Cutis Total Triterpenoids Tablets Based on Liquid-solid Compact Technique

[Objectives]To study the immediate-release mechanism of dehydrotrametenolic acid and dehydroeburicoic acid in Poriae Cutis total triterpenoids tablets by liquid-solid compacts technique.[Methods] Taking dehydrotrametenolic acid and dehydroeburicoic acid as indicators,differences in dissolution were compared between liquid-solid compressed tablets and crude drug of total triterpenoids in Poriae Cutis;crude drug,powder of liquid-solid compressed tablets and excipients of liquid-solid compressed tablets were characterized by differential scanning calorimetry( DSC).[Results]Liquid-solid compact technique could significantly improve dissolution rate of dehydrotrametenolic acid and dehydroeburicoic acid. The total dissolution rate of dehydrotrametenolic acid and dehydroeburicoic acid in Poriae Cutis total triterpenoids tablets was 92%,t50( time for 50% total dissolution rate) and t_D( time for 63. 2% total dissolution rate) were 11. 18 min and 22. 71 min; the total dissolution rate of dehydrotrametenolic acid and dehydroeburicoic acid in crude drug of total triterpenoids was 29%,and t50 and t_D were231. 06 min and 359. 23 min. DSC showed that there was no mutual interaction between excipients and total triterpenoids in Poriae Cutis; on the DSC curve for powder of liquid-solid compressed tablets,the absorption peak vanished completely,indicating that the drug exists in the amorphous form in the liquid-solid powder.[Conclusions] Liquid-solid compact technique can increase the dissolution rate of total triterpenoids in Poriae Cutis and allow rapid release of poorly water soluble drugs.

Effects of Different Decolorants on Retention Rate of Total Triterpenes in Fruit and Rattan Stems of Schisandra chinensis(Turcz.) Baill

This study was to investigate the decoloration effects of activated clay, activated carbon, diatomite, Ca group bentonite, kaolin, activated aluminum oxide, magnesium oxide and attapulgite. Ultraviolet-visible spectroscopywas adopted to de- tect the absorbanees of the extracts of Schisandra chinensis fruit and rattan stems, the contents of total triterpenoids in S. chinensis fruit and rattan stems were deter- mined, and the decoloration rate and the retention rate of total triterpenoids of S. chi- nensis fruit and rattan stems were calculated. The results indicated that, attapulgite exhibited the best decoloration effect on the extract of S. chinensis fruit, with a de- coloration rate up to 60.47%, activated carbon exhibited a better decoloration effect on the extract of S. chinensis rattan stems, with a decoloration rate up to 69.24%, and they had relatively higher retention rates of total triterpenoids. In the eight de- colorants, attapulgite showed the best decoloration effect on the extract of S. chi- nensis fruit, and activated carbon exhibited the best decoloration effect on the ex- tract of S. chinensisrattan stems.

响应面法优化提取瓜蒌皮中总三萜的研究

目的 探讨用醇提法提取瓜蒌皮中总三萜化合物。方法 以瓜蒌皮为研究对象,以乙醇为溶剂,采用紫外分光光度法对瓜蒌皮中总三萜化合物的含量进行测定。通过单因素实验研究液固比、提取温度、提取时间、乙醇浓度对总三萜提取率的影响,在单因素实验的基础上,采用Box-Behnken Design响应面分析法研究乙醇提取的最佳工艺条件。结果 冷凝回流提取的最佳条件为液固比51 mL/g,乙醇浓度83%、提取时间70 min、提取温度90℃;在此工艺条件下,总三萜的最佳提取率为2.324%。结论 研究结果与预测值基本一致,因此该模型可靠。

响应面法优化超声辅助提取蕨麻总三萜工艺研究

目的:优化超声辅助提取蕨麻总三萜工艺条件研究。方法:在单因素试验基础上,利用响应面法评估液料比、提取时间、提取温度对总三萜提取得率的影响。结果:超声辅助提取最佳工艺参数为液料比20∶1 mL/g,提取时间63 min,提取温度60℃,蕨麻总三萜提取得率可达4.73%±0.01%。结论:响应面法优化提取工艺切实可行,操作简单,总三萜含量高,可为蕨麻进一步开发利用提供依据。

凌霄花总三萜的超声辅助提取工艺优化

目的 优化凌霄花总三萜类化合物的超声辅助提取工艺。方法 以总三萜提取率为评价指标,考察料液比、乙醇浓度、超声时间、超声功率等因素对提取率的影响,采用单因素试验和正交试验优化提取工艺。结果 凌霄花总三萜的最佳提取工艺为:料液比1/40(g·mL^(-1)),乙醇浓度60%,超声时间40min,超声功率100W。该条件下,凌霄花总三萜平均提取率为4.11%。结论 本研究优化了凌霄花总三萜的超声辅助提取工艺,该工艺稳定,提取效率较高,适合该药材三萜类化学成分的提取。

TT、TSI、FTI评估男性雄激素水平价值的初步研究

目的:游离睾酮(FT)是有活性的睾酮,可以准确反应男性雄激素水平,是判定男性雄激素缺乏最可靠的检验指标,但是检测方法复杂,所以目前常用计算游离睾酮(cFT)替代。血清总睾酮(TT)、睾酮分泌指数(TSI)、游离睾酮指数(FTI)是3种常用的判定男性雄激素缺乏的指标。通过研究上述指标与cFT的相关性,探讨TT、TSI、FTI评估男性雄激素水平的价值。方法:选取大连医科大学附属第一医院男科门诊2012年1月至2012年12月就诊的男科患者161例,测定患者血清总睾酮(TT)、黄体生成素(LH)、性激素结合蛋白(SHBG)和白蛋白(ALB),通过Vermeulen公式计算cFT、睾酮分泌指数(TSI=TT/LH)、游离睾酮指数(FTI=TT/SHBG)。以TT≤11.5nmol/L、TSI≤2.8nmol/IU、FTI≤0.42nmol/nmol为参考截点值,cFT≤0.3nmol/L为标准,探讨TT、TSI、FTI评估男性雄激素水平的价值。结果:以cFT为评估标准,评估男性雄激素水平的符合率分别为:TT 84.9%、TSI 74.0%、FTI 71.1%。诊断男性雄激素缺乏的漏诊率分别为TT 33.3%、TSI 24.6%、FTI 62.3%,诊断男性雄激素缺乏的误诊率分别为TT 2.1%、TSI 26.9%、FTI 5.2%。Kappa值分别为:TT 0.676、TSI 0.472、FTI 0.411。P值均小于0.01,具有统计学意义。结论:(1)TT、TSI、FTI无法替代cFT,准确评估男性雄激素水平仍需检测cFT。(2)TT、TSI在初步评估男性雄激素水平方面有参考价值:①TT评估男性雄激素缺乏的准确性好;②TSI可作为筛查男性雄激素缺乏的指标。(3)相对于TT、TSI,FTI在评估男性雄激素水平方面价值较小。

牛樟芝总三萜提取工艺优化及其抗氧化活性研究

牛樟芝是我国台湾特有的珍稀药食两用真菌,含有丰富活性成分,具有抗癌、抗肿瘤和保肝等多种药理作用,三萜类化合物被认为是牛樟芝中的重要药用物质,不同提取工艺对牛樟芝总三萜得率影响不同.为探究牛樟芝总三萜最佳提取工艺及其抗氧化活性.选取不同提取溶剂、温度、时间、液料比,采用单因素试验方法和正交试验方法探究不同因素对牛樟芝总三萜得率的影响,并通过高效液相色谱(HPLC)分析不同溶剂提取工艺之间的成分差异,比较分析其抗氧化活性.结果表明,牛樟芝总三萜提取最佳工艺为:二氯甲烷为提取溶剂、液料比1∶320 g·mL^(-1)、45℃提取5 h.在此条件下,牛樟芝总三萜得率为10.16±0.077 6%,通过HPLC分析,所得粗提物成分丰富且具有较好的抗氧化活性,其中DPPH清除能力的IC50值达0.137 6,总还原能力达58.03%.通过对牛樟芝提取工艺优化显著提高了牛樟芝总三萜得率,为牛樟芝活性成分提取、化合物分离及开发利用提供理论参考.

星点设计-效应面法优化超声提取桂籽中三萜化合物的工艺研究

目的本研究以桂籽为原材料,采用星点设计(CCD)-效应面法(RSM)等多种方法优化桂籽总三萜(OFSTT)的提取工艺及其含量测定研究。方法采用单因素、Plackett-Burman(PB)、最陡爬坡和CCD-RSM实验,建立各因素与响应值之间的数学模型,确定OFSTT的最佳超声提取工艺。结果通过单因素试验,得到了关于桂籽总三萜含量变化的趋势线,进一步利用PB实验筛选出对OFSTT产生显著影响的因素(提取功率、液料比及提取次数),通过最陡爬坡实验设计出极值点,以此作为效应面设计的中心点,采用星点设计-效应面法优化了OFSTT提取工艺的最佳提取工艺条件:乙醇溶剂分数60%,液料比25∶1(mL/g),提取温度65℃,提取功率325W,提取时间30min,提取3次;该条件下OFSTT的提取率为7.251%,实际值与预测值的偏差为2.05%。结论本研究采用CCD-RSM实验优化了OFSTT的提取工艺,该工艺稳定,提取率较高,实际值与预测值吻合度高,为后续桂籽三萜类物质进一步结构鉴定及生物活性作用研究提供参考价值。

Total triterpenes from fruits of Chaenomeles speciosa(Sweet) Nakai protect against indomethacin-induced gastric mucosal injury: involvement of TFF1-mediated EGF/EGFR and apoptotic pathways

OBJECTIVE Gastric ulcers affect people of all ages and half of the world's population,which is being considered as the new"plague of the 21st century".As is well known,gastric mucosa is known as the first guard to protect the stomach from ulcer injury,while the aetiology of gastric ulcer is relative to imbalances between gastric mucosal protective and aggressive factors.Therefore,reducing or eliminating the aggressive factors,returning to the balance of between mucosal protective and aggressive factors,and then restoring the normal functional of gastric mucosal barrier could be crucial for treating the gastric ulcer.The fruits of Chaenomeles speciosa(TCS),also known as"mugua",might be processed into edible and health care derived products,and used as a commonly used traditional medicine in China for thousands of years.In China folk,there is a saying that"apricot one benefit,pear two benefits,mgua hundred benefits",so it has a"hundred-benefit"fruit reputation.Tujia nationality inhabitants in Southwestern China should have rheumatic diseases and peptic ulcers for living in the damp environments and bingeing on spicy and pungent foods.For the exis⁃tence of the fruit derived products of Chaenomeles speciosa as their complementary foods or snacks,the habit makes them rarely suffer from the two kinds of diseases.Enlightened by these,we had investigated the structure-activity rela⁃tionships,screened out CSTT with gastroprotective activity.Our previous studies demonstrated that TCS owned effec⁃tively therapeutic effects on gastric ulcer patients and animals,and further confirmed that TFF1 and apoptotic pathway were closely interrelated with its exerting gastroprotection.However,its underlying molecular mechanisms involved have not been fully elucidated.The current study was to further investigate its protective effect on indomethacin(IND)-damaged RGM-1 cells and rats and its underlying mechanisms through modulating TFF1-mediated EGF/EGFR and apoptotic pathways.METHODS The gastroprotection of TCS was evaluated with IND-induced gastric lesions model in RGM-1 cells and rats.In vitro,the proliferation,migration,mitochondrial viability and apoptosis were assessed,In vivo,ulcer index,ulcer inhibition rate,gastric juice acidity,gastric wall mucus(GWM),histopathology of gastric mucosa were detected.The gastroprotective effects of TCS through the TFF1-mediated EGFR/EGFR and apoptotic pathways were presented and measured by qRT-PCR and Western blotting assays.RESULTS TCS had gastroprotective function,which was related to the amelioration in promoting IND-damaged RGM-1 cell proliferation and migration,hoisting gastric juice acidity and GWM,improving ulcer index and ulcer inhibition rate,attenuating the hemorrhage,edema,epithelial cell loss and inflammatory cell infiltration of gastric mucosa,upregulating proliferation cell nuclear antigen,Bcl-2,Bcl-xl mRNA and TFF1,EGF,p-EGFR,p-Src,pro-caspase-3,pro-caspase-9 protein expressions,mitochondrial viability,mitochondrial cytochrome C concentration and p-EGFR/EGFR,p-Src/Src,Bcl-2/Bax,Bcl-xl/Bad ratioes,downregulating Bax,Bad,Apaf-1 mRNA and cleaved-caspase-3,cleaved-caspase-9,cleaved-PARP-1 protein expressions,cytosol cytochrome C concentration.CONCLUSION TCS′s gastroprotective effect was closely connected with boosting TFF1 expression,acti⁃vating TFF1-mediated EGF/EGFR pathway,thus restraining mitochondrial-dependent apoptosis,which provided new insights into interpreting its underlying mechanism and promised to act as a candidate drug to treat gastric mucosal injury.

血清睾酮、雌二醇及睾酮/雌二醇比值在生精功能障碍评估中的应用价值分析

目的 分析血清睾酮、雌二醇(E_(2))及睾酮/E_(2)比值在生精功能障碍评估中的应用价值。方法 选择2020年6月至2021年12月于广西壮族自治区妇幼保健院生殖中心就诊的750例男性的临床资料。其中精液质量正常者300名(正常对照组),生精功能障碍者450例。根据精子总数,生精功能障碍者分为轻度少精组、重度少精组和非梗阻性无精子症组,每组150例。分析精液质量正常者性激素指标水平的百分位数分布情况。比较四组血清促卵泡刺激素(FSH)、总睾酮(TT)、E_(2)、游离睾酮(FT)、TT/E_(2)比值和FT/E_(2)比值。结果 精液质量正常男性TT和FT的均值分别为504.50 ng/dl和10.28 ng/dl,第5百分位数值分别为273.28 ng/dl和6.70 ng/dl;TT/E_(2)比值和FT/E_(2)比值的均值分别为15.75和0.32,第20百分位数值分别为10.06和0.23。随着生精障碍严重程度加重,FSH水平呈上升趋势,E_(2)、TT和FT水平呈下降趋势。非梗阻性无精子症组TT、FT、E_(2)水平以及TT/E_(2)比值、FT/E_(2)比值与正常对照组比较差异均有统计学意义(P<0.05)。非梗阻性无精子症组FSH水平显著高于轻度少精组和重度少精组(P<0.05),FT水平、FT/E_(2)比值显著低于轻度少精组和重度少精组(P<0.05)。与正常对照组和轻度少精组相比,重度少精组FSH水平升高,TT、FT水平以及TT/E_(2)比值、FT/E_(2)比值降低,差异有统计学意义(P<0.05)。轻度少精组FSH水平高于正常对照组,FT水平低于正常对照组,差异有统计学意义(P<0.05)。结论 生精功能障碍评估中血清FT是值得关注的重要指标,TT和FT参考值下限分别为273.28 ng/dl和6.70 ng/dl;TT/E_(2)比值<10.06可作为治疗界限值。

石胡荽醇提物制备工艺优化及其不同萃取部位抗炎活性研究

目的 优化石胡荽醇提物制备工艺,并考察其不同萃取部位的抗炎活性。方法 采用单因素实验和响应面法,以石胡荽总三萜提取率为评价指标,考察乙醇体积分数、提取时间、料液比、提取次数4个因素对总三萜醇提物加热回流提取工艺的影响,优化最佳工艺,并进行验证实验。以地塞米松为阳性对照药物,比较石胡荽不同萃取部位(石油醚部位、乙酸乙酯部位、正丁醇部位、水部位)对脂多糖诱导小鼠单核巨噬细胞RAW 264.7一氧化氮(NO)生成的影响并计算其半数抑制浓度(IC50)。结果 石胡荽总三萜醇提物的最优提取工艺为乙醇体积分数70%、料液比1∶40(g/mL)、提取时间2.0 h、提取次数3次。3次验证实验表明,石胡荽总三萜的平均提取率为1.134%,与预测值的相对误差为0.02%。石胡荽的石油醚部位、乙酸乙酯部位可不同程度地降低由脂多糖诱导的RAW 264.7细胞的NO生成,其对NO生成的IC50分别为2.44、2.22μg/mL,均低于阳性对照药物地塞米松(7.65μg/mL)。结论 优化后的石胡荽醇提物的制备工艺稳定可行,其石油醚部位、乙酸乙酯部位具有较好的抗炎活性。

几种市售野生食用菌中3种活性物质含量的测定

以市售11种野生食用菌为研究对象,采用分光光度法测定粗多糖、总三萜含量,高效液相色谱法测定腺苷含量。结果表明,11种野生食用菌的粗多糖含量为1.44~5.27 g·100-1·g^(-1),总三萜含量为0.75%~1.60%,腺苷含量为657~3349 mg·kg^(-1)。其中,白牛肝菌(Boletus bainiugan)的粗多糖和腺苷含量最高,香肉齿菌(Sarcodon aspratus)的总三萜含量最高。3种活性物质的变异系数从高到低依次为腺苷、粗多糖、总三萜。

响应面法优化阿里红总三萜酸提取工艺及抗氧化活性研究

以6个不同产地的阿里红为研究对象,采用单因素实验结合响应面法对超声辅助提取阿里红总三萜酸(FOATTA)工艺进行优化,通过测定其对DPPH自由基清除率及Fe^(3+)还原能力考察其抗氧化活性。确定最佳提取工艺为:料液比1∶33(g∶mL)、乙醇体积分数85%、提取时间40 min,在此条件下,FOATTA得率为14.510%;在实验浓度范围内,随着浓度的增加,不同产地FOATTA对DPPH自由基清除能力和Fe^(3+)还原能力均呈上升趋势,表明其具有一定的抗氧化能力。优化后的超声辅助提取工艺可行性强,操作简便,为进一步研究和开发阿里红资源提供了参考。

响应面法优化地皮菜总三萜提取工艺及其抗氧化活性

以青海地皮菜为原料,研究其总三萜提取工艺条件及其抗氧化活性,通过单因素试验和Box-Behnken响应面试验进行超声波辅助提取地皮菜总三萜工艺优化,并测定地皮菜总三萜对DPPH自由基、ABTS^(+)自由基、羟自由基的清除能力及总还原能力,以评估其体外抗氧化活性。最终得出的最佳提取条件为料液比1∶20(g/mL)、乙醇浓度95%、提取温度60℃、提取时间50 min,在此条件下测得地皮菜总三萜的得率为1.38%,与模型预测值1.41%基本相符。抗氧化试验结果表明,当地皮菜总三萜质量浓度为1.60 mg/mL时,DPPH自由基清除率达到最大,为48.43%;ABTS^(+)自由基清除率随质量浓度的增加而增加,当浓度为3.20 mg/mL时,ABTS^(+)自由基清除率达到最大,为81.43%;羟自由基清除率与浓度无明显线性关系;地皮菜总三萜的总还原能力随着浓度的增加呈增大趋势,当浓度为3.20 mg/mL时,总还原能力达到1.818。以上结果表明,响应面法对地皮菜总三萜的提取条件优化合理可行,且地皮菜总三萜具有较高的抗氧化活性。

大孔树脂纯化灵芝子实体和孢子粉中总三萜

研究比较大孔树脂对灵芝子实体和孢子粉两个不同部位中总三萜的吸附特性,优选其对灵芝总三萜的分离纯化工艺条件。采用紫外分光光度法测定灵芝子实体和孢子粉两个不同部位中总三萜的含量,选择8种大孔树脂,以总三萜吸附率和解吸率为指标,筛选最佳树脂型号,并考察最佳树脂纯化灵芝子实体和孢子粉中总三萜的工艺参数。结果表明:8种大孔树脂中,HPD-500型大孔树脂对灵芝子实体总三萜的吸附解吸效果最好,最佳纯化工艺条件为上样液浓度1.0 g干样/mL,最大上样量2 BV,用8.5 BV的75%乙醇进行洗脱,该工艺下灵芝子实体总三萜纯度达60.59%;而AB-8型大孔树脂对灵芝孢子粉总三萜的吸附解吸效果最好,且最佳纯化工艺条件为上样液浓度0.4 g干样/mL,最大上样量3 BV,用8.5 BV的95%乙醇进行洗脱,该工艺下灵芝孢子粉总三萜纯度达65.48%。综上,HPD-500型和AB-8型大孔树脂分别富集纯化灵芝子实体和灵芝孢子粉中总三萜的工艺效果显著,简单可行,为灵芝三萜的深入探究和工业化纯化灵芝子实体和孢子粉总三萜提供理论依据。

龙牙楤木Aeβ-AS基因的表达对烟草中皂苷含量的影响

为探讨龙牙楤木(Aralia elata)三萜皂苷合成的关键酶基因β-香树素合成酶基因(Aeβ-AS)对三萜皂苷合成的影响。利用基因克隆技术对Aeβ-AS基因进行克隆,并对烟草进行遗传转化,分析转基因烟草中Aeβ-AS基因在各部位的表达差异,检测Aeβ-AS基因及上下游关键酶基因表达量和总三萜含量。结果表明:成功构建了植物过表达载体pROKⅡ-Aeβ-AS,并转入了野生型烟草,获得7个转基因株系,并在mRNA水平表达,且在叶子中的表达量要高于根和茎;在转基因烟草中,Aeβ-AS基因及其上下游的关键酶基因表达量均高于野生型,其中株系L21的NtFPS、NtSS基因的相对表达量最高,株系L30的NtSE、Aeβ-AS基因的相对表达量最高;与野生型烟草相比,转基因烟草的总三萜含量显著升高(1.1~1.6倍)。试验结果证明合成Aeβ-AS基因并在烟草中进行异源转化,可以使转基因烟草内的总三萜含量显著升高。

瑶药威灵仙总三萜超声辅助提取工艺优化以及抗氧化抗菌活性的初步研究

本研究以瑶药威灵仙为研究对象,采用单因素实验结合响应面法对超声辅助提取威灵仙总三萜工艺进行优化;通过测定威灵仙总三萜还原Fe^(3+)-TPTZ产生Fe^(2+)-TPTZ的能力考察其总抗氧化活性;通过测量威灵仙总三萜对金黄色葡萄球菌和大肠杆菌的抑制圈的面积考察其抗菌活性。确定最佳提取工艺为:乙醇体积分数55%、提取时间35 min,温度35℃。在此条件下,瑶药威灵仙总三萜得率为1.280%(W/W);在实验浓度范围内,随着浓度的增加,威灵仙总三萜对Fe^(3+)-TPTZ的还原能力呈上升趋势,表明其抗氧化能力与浓度具有依赖性;其对金黄色葡萄球菌和大肠杆菌的抑制能力随浓度的增加上升不明显,说明其抗菌活性的浓度依赖性不强。

鹿角菜醇提物不同极性部位活性成分及其体外抗氧化活性研究

研究鹿角菜醇提物不同极性部位总黄酮、总多酚和总三萜质量浓度及其抗氧化活性关系。以鹿角菜为主要原料,采用标准曲线法计算鹿角菜醇提物各不同萃取相中总黄酮、总多酚和总三萜的含量,同时通过ABTS^(+)·法、DPPH·法、O_(2)^(-)·法和·OH法以及Fe^(3+)还原法,计算不同萃取相下的鹿角菜醇提物体外抗氧化能力。结果表明,鹿角菜醇提物中黄酮类物质含量最高,三萜类物质含量最低。通过不同极性部位比较发现,鹿角菜醇提物乙酸乙酯相中抗氧化活性物质总含量最高,抗氧化能力最强,其对ABTS^(+)·、DPPH·、O_(2)^(-)·及·OH的清除率及Fe^(3+)的还原性分别为98.17%、96.89%、31.99%、100%和1.1297。由此可见,鹿角菜醇提物的乙酸乙酯相可作为鹿角菜后期研究的核心。