In Vitro Cytotoxicity and Anti-tumor Properties of the Total Alkaloid Fraction of Unripe Fruits of Solanum pseudocapsicum

The total alkaloid fraction of the methanol extract of unripe fruits of Solanum pseudocapsicum was tested for in vitro cytotoxicity on HEp-2, RD and Vero cell lines and anti-tumor activity using DLA and HEp-2 cell lines. Cell viability and morphological changes were assessed. The total alkaloid fraction exhibited strong cytotoxic activity against all the cell lines tested. The CTC 50 (50 % of cytotoxicity inhibition) was found to be 1.65 µg/ml for the RD cell line, 6.32 µg/ml for the HEp-2 cell line and 12.01 µg/ml for the Vero cell line. In the clonogenic assay, no colony formation was observed even up to a concentration of 25 µg/ml. In the short term, antitumor, studies using DLA cells, the total alkaloid fraction was associated with 50 % viability in the concentration range of 6.25-12.5 µg/ml. In long term, anti-tumor activity using the HEp-2 cell line, no colony formation was observed up to a concentration of 20 µg/ml. Hence, there is a correlation between the results obtained in the cytotoxicity and antitumor studies carried out. Morphological observation by phase contrast microscopy revealed intense damage on all the cell lines. The total alkaloid fraction has the potential for further investigation in animal models.     

刺山柑总生物碱乳膏外用对系统性硬皮病小鼠组织纤维化的改善作用

目的 研究刺山柑总生物碱乳膏外用对系统性硬皮病(systemic sclerosis,SSc)小鼠组织纤维化的改善作用。方法 BALB/c小鼠背部注射盐酸博莱霉素建立SSc模型,外用刺山柑总生物碱乳膏进行干预后,比较给药前后小鼠真皮厚度的变化,ELISA法检测小鼠皮肤及肺组织羟脯氨酸(Hyp)和Ⅰ型胶原(Col-Ⅰ)表达水平。结果 刺山柑总生物碱乳膏中、高剂量可明显降低SSc小鼠真皮厚度,同时可降低小鼠皮肤及肺组织Hyp含量(P<0.05或0.01),高剂量还可降低肺组织Col-Ⅰ含量(P<0.01)。结论 刺山柑总生物碱乳膏对SSc组织纤维化有一定改善作用。     

Free Radical Scavenging and Hepatoprotective Constituents from the Leaves of Juglans sinensis

In the course of searching for hepatoprotective agents from natural products, six compounds were isolated from the MeOH extract of the leaves of Juglans sinensis, as guided by their DPPH free radical scavenging activity. The structures were determined as juglanoside B (1), quercetin 3-O-alpha-L-arabinofuranoside (avicularin, 2), quercetin 3-O-alpha-L-arabinopyranoside (guaijaverin, 3), quercetin 3-O-alpha-L-rhamnopyranoside (quercitrin, 4), (+)-catechin (5) and quercetin 3-O-beta-D-galactopyranoside (hyperin, 6). Compounds 2-6 showed significant DPPH free radical scavenging effects. An evaluation for the hepatoprotective activity of the isolated compounds on drug-induced cytotoxicity was conducted, and compounds 1, 2, and 5 showed protective effects against nitrofurantoin-induced cytotoxicity, and compound 5 also exhibited a moderate protective effect on amiodarone-induced cytotoxicity in Hep G2 cells.     

川芎中总生物碱、总酚酸和总挥发油对大鼠离体子宫平滑肌收缩活动的抑制作用

目的:考察川芎中总生物碱、总酚酸和总挥发油对大鼠离体子宫平滑肌收缩活动的抑制作用,为阐明川芎活血调经的传统功效提供依据。方法:取大鼠离体子宫平滑肌浸泡于洛氏液中,以二甲基亚砜为空白对照、盐酸维拉帕米为阳性对照,采用Power Lab生理记录仪记录子宫平滑肌收缩曲线,通过观察收缩活动力、收缩张力变化来评价川芎中总生物碱、总酚酸(质量浓度均分别为0.025、0.05、0.1 mg/m L)和总挥发油(0.04、0.08、0.16 mg/m L)对缩宫素致大鼠离体子宫平滑肌收缩的抑制作用,并计算子宫平滑肌收缩活动力抑制率和收缩张力抑制率。结果:加入缩宫素后大鼠离体子宫平滑肌的收缩活动力、收缩张力较加入缩宫素前均显著升高(P<0.05或P<0.01),不同质量浓度3类川芎组分均可显著降低子宫平滑肌的收缩活动力、收缩张力(P<0.05或P<0.01)。与空白对照比较,除0.025 mg/m L总酚酸外,给予其余浓度药物或组分后子宫平滑肌的收缩活动力抑制率和收缩张力抑制率均显著升高(P<0.05或P<0.01)。结论:川芎中总生物碱、总酚酸和总挥发油对缩宫素致大鼠离体子宫平滑肌收缩均有一定的抑制作用,这为川芎活血调经的用法提供了参考依据。     

单胺氧化酶抑制剂氯吉灵抑制结肠癌的作用及机制研究

目的：研究单胺氧化酶A （monoamine oxidase A,MAO-A）抑制剂氯吉灵（Clorgyline）对结肠癌细胞增殖、转移的作用,以及其对MAO-A的酶活、MAO-A、MMP-2和MMP-9蛋白表达的影响。方法：MTS法检测不同浓度Clorgyline对结肠癌细胞SW480增殖的作用;划痕实验研究Clorgyline对SW480细胞迁移的影响;Transwell实验研究Clorgyline对SW480细胞侵袭的影响;裸鼠移植瘤模型研究Clorgyline对SW480细胞体内增殖的抑制作用;酶活试剂盒检测Clorgyline对裸鼠移植瘤组织中MAO-A的作用;Western blot检测Clorgyline对裸鼠移植瘤组织中的MAO-A、MMP-2和MMP-9蛋白表达的影响。结果：Clorgyline对SW480细胞增殖有抑制作用,且呈现剂量依赖性;Clorgyline 10 μmol·L^-1和20 μmol·L^-1浓度均能够抑制SW480细胞迁移和侵袭能力,与对照组相比具有显著性差异（P<0.01）;Clorgyline 20和40 mg·kg^-1均能够抑制SW480细胞裸鼠移植瘤的生长（P<0.01）,抑制裸鼠移植瘤组织中MAO-A的酶活（P<0.05）,抑制MMP-2和MMP-9蛋白的表达水平,而对MAO-A蛋白的表达水平没有影响。结论：Clorgyline抑制SW480结肠癌细胞的增殖和转移,其机制可能与抑制MAO-A酶活和转移相关蛋白MMP-2、MMP-9的表达有关。     

基于LC-MS树参叶指纹图谱的建立

目的根据树参叶液相色谱-质谱联用的分析结果,建立其液相色谱指纹图谱的鉴别方法。方法采用Waters BEH C18色谱柱(2.1 mm×100 mm,1.7μm)。流动相:乙腈(A)-0.1%甲酸(B),梯度洗脱,流速0.2 mL·min^-1,柱温35℃,进样量为1μL;电喷雾电离源(ESI),负离子检测,离子源温度为150℃,脱溶剂气温度为500℃,脱溶剂气流氮气800 L·h^-1,锥孔气流氮气50 L·h^-1,碰撞气为氩气,毛细管电压为3000 V,对树参叶主要成分进行确证。采用Waters SunfireTM C18色谱柱(4.6 mm×250 mm,5μm),流动相:乙腈(A)-混合水溶液(含甲酸0.2%和四氢呋喃0.06%)(B),梯度洗脱,检测波长256 nm,柱温30℃,体积流量1.0 mL·min^-1,分析时间80 min,进样量为5μL,建立树参叶的指纹图谱鉴别方法。结果10批样品相似度为0.939~0.989,均符合要求。结论树参为常绿植物,其叶中所含化学成分和植物的生长年限、环境、采收季节密切相关,建立的指纹图谱鉴别方法简便、重复性好,可为树参叶的品质评价提供科学依据。     

高山红景天总多酚对高脂食物-链脲佐菌素诱导大鼠2型糖尿病的影响

目的:研究高山红景天总多酚对高脂食物-链脲佐菌素诱导2型糖尿病大鼠高血糖的保护作用。方法:36只大鼠均分为6组,即正常对照(等容生理盐水)、模型(等容生理盐水)、二甲双胍(0.3g.kg-1)和高山红景天总多酚高、中、低剂量(100、50、25g.kg-1)组,ig给药,每天1次,连续21d。末次给药12h后尾静脉取血,测血糖值,禁食不禁水18h后,测定胰腺线粒体中超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,评定高山红景天总多酚的降糖作用。结果:模型组血糖值显著高于正常对照组(P<0.01);与模型组比较,高山红景天总多酚高、中、低剂量组大鼠血糖值显著降低(P<0.01),且中剂量组与二甲双胍组比较无显著性差异。与正常对照组比较,模型组大鼠胰腺线粒体中MDA含量显著升高(P<0.01),SOD活性显著降低(P<0.01);与模型组比较,高山红景天总多酚高、中、低剂量组MDA含量显著降低(P<0.01),SOD活性显著升高(P<0.01)。结论:高山红景天总多酚具有良好的降糖作用,其机制很可能是高山红景天多酚类成分的抗氧化作用。     

壳多糖保肝作用的实验研究

目的：观察壳多糖(chitosan)保肝的药理作用。方法：按200 mg·kg 1的剂量给予小鼠灌胃给药，观察壳多糖对四氯化碳致小鼠急性肝损伤的组织学变化及其对转氨酶、脂质过氧化的影响。结果：壳多糖组小鼠肝细胞变性、坏死明显轻于造模组；壳多糖组ALT水平，血浆、肝匀浆过氧化脂质(LPO)及血浆丙二醛(MDA)含量亦显著低于造模组，组间比较差异具有极显著性或显著性(P＜0.01或P＜0.05)。结论：壳多糖对小鼠急性实验性肝损伤有明显的保肝降酶作用，机制可能与其抗自由基损伤、抑制脂质过氧化反应等有关。     

不同产地桑叶氨基酸、核苷、生物碱成分的含量测定及多元统计分析

目的 建立超高效液相串联三重四极杆质谱（UPLC-TQD）同时测定桑叶样品中氨基酸、核苷及生物碱[1-脱氧野尻霉素（1-deoxynojirimycin,DNJ）]含量的方法,并采用多元统计分析方法分析比较不同产地桑叶中成分含量差异,为桑叶资源的品质评价提供科学依据。方法 采用Waters Acquity UPLC BEH Amide（100 mm×2.1 mm,1.7 μm）色谱柱,以0.2%甲酸乙腈（A）-0.2%甲酸水（B）梯度洗脱,流速0.4 mL·min^-1,柱温35℃;使用多反应监控正离子模式测定;采用多元统计分析如主成分分析对不同产地桑叶进行综合性评价。结果 22种成分的浓度与峰面积呈良好的线性关系（r² ≥ 0.990 3）;方法精密度、重复性及稳定性的RSD值均≤ 7.5%;平均加样回收率为86.7%~106.4%,RSD为2.2%~8.9%。不同产地桑叶中氨基酸、核苷及DNJ存在一定差异,结合多元统计分析,共找出8个差异性成分,可将样品分成2组。结论 所建立方法准确可靠,结合DNJ及8个差异性成分含量进行比较,湖北及江西产地的桑叶品质较优。     

Antioxidant Activities of the Chloroform Extract of Solanum trilobatum

A chloroform extract from the anticancer herb Solanum trilobatum (CST) (Nadkarni, 1979) was analyzed and compared with reference antioxidants for its in vitro antioxidative properties such as scavenging of α,α-diphenyl-β-picryl hydrazyl (DPPH) and superoxide radicals, protection to deoxyribose degradation, reducing power, as well as inhibition of lipid peroxidation. Preliminary phytochemical analysis of CST by silica gel thin-layer chromatography showed the presence of simple phenols, phenolic acids, isoflavones, xanthones, and lignans. The antioxidative effect of CST was found to be concentration dependent to a certain extent and then leveled off with further increase in concentration. The IC₅₀ for each antioxidative reaction studied was calculated. When compared to the reference antioxidant butylated hydroxytoluene (BHT), CST exhibited less scavenging effect on DPPH radicals and reducing power but a better superoxide radical scavenging effect. From a comparison of the hydroxyl radical scavenging effect of CST with catechin, it seemed that CST was four-times more effective than catechin. CST was also able to prevent the formation of ·OH-induced malondialdehyde (MDA) in rat liver homogenate.     

Evaluation of Acute and Subacute Toxicity and LC-MS/MS Compositional Alkaloid Determination of the Hydroethanolic Extract of Dysphania ambrosioides (L.) Mosyakin and Clemants Flowers

Dysphania ambrosioides (L.) Mosyakin and Clemants is a medicinal plant that has traditionally been used to cure a range of diseases. There has been no thorough investigation of the potential toxicity of this plant. The objective of this study is to assess the acute and subacute toxicity of D. ambrosioides hydroethanolic extract (DAHE), as well as it alkaloids composition, utilizing LC-MS/MS analysis. An in silico approach was applied to determine pharmacokinetic parameters and to predict the toxicity of D. ambrosioides identified alkaloids. A 14-day treatment with a single oral dose of 1–7 g/kg was carried out to investigate acute toxicity. DAHE was given orally at dosages of 5, 50, and 500 mg/kg for 15 days in the subacute toxicity investigation, and body weight and biochemical parameters were evaluated. Livers, kidneys, lungs, and heart were examined histologically. Chromatographic investigation revealed the existence of nine alkaloids, with N-formylnorgalanthamine being the most prevalent. The oral LD₅₀ value of DAHE was found to be 5000 mg/kg in an acute toxicity study. No variations were observed with respect to food intake, water consumption, mortality, or body and organ weight in the subacute toxicity study. On the other hand, DAHE (500 mg/kg) significantly enhanced alanineaminotransferase, aspartate aminotransferase, and urea. Liver and kidney histological examinations revealed modest infiltration of hepatocyte trabeculae by inflammatory cells in the liver and slight alteration in the kidney histoarchitecture. According to our findings, DAHE exhibits low to moderate toxicity.     

正交试验法筛选骆驼蓬总碱片处方的实验研究

目的 :优选骆驼蓬总碱片最佳处方。方法 :以辅料碳酸钙、吐温 -80、淀粉及润滑剂 (硬脂酸镁∶滑石粉 )用量为4因素 ,每因素取3水平 ,以总碱片中去氢骆驼蓬碱的体外溶出参数 (Td)为指标进行正交试验。结果 :辅料中影响去氢骆驼蓬碱Td 的主要因素为吐温 -80和碳酸钙用量 (P<0 .05) ;经试验筛选出的最佳处方为骆驼蓬总碱浸膏粉适量 (总碱10mg/片 )、淀粉45mg、吐温 -80用量1 %、硬脂酸镁0. 1mg、滑石粉0 .31mg、氧化镁适量 ;Td=25 .01min。结论 :筛选出的最佳处方具有辅料价廉易得、有效成分溶出快、制备简单、稳定等特点。     

墨旱莲提取物对肝保护作用的影响

为研究采用不同方法提取的墨旱莲提取物是否对其肝保护作用有影响 ,预先分别给予小鼠口服干品墨旱莲全草的水煎剂、5 0 %乙醇提取物及乙酸乙酯提取物的水溶物 ,连续 7天后 ,腹腔注射醋氨酚诱发小鼠急性肝损伤 ,测定血清谷氨酸氨基转移酶(s ALT)、血清天冬氨酸氨基转移酶 (s AST)的活力。结果显示 ,墨旱莲各组提取物均能显著地抑制醋氨酚诱发的小鼠 s AL T、s AST升高 ,与醋氨酚损伤组比较 ,乙酸乙酯提取物与 5 0 %乙醇提取物分别有非常显著差异 (P<0 .0 1)和显著差异 (P<0 .0 5 ) ,而水煎剂则几乎没有肝保护作用 (P>0 .0 5 )。提示采用不同的提取方法得到的墨旱莲提取物对醋氨酚诱发的小鼠急性肝损伤有不同程度的保护作用 ,其中以乙酸乙酯提取物的效果最显著     

不同产地哈萨克药材白喉乌头中总生物碱含量的分析

目的 测定不同产地白喉乌头所含总生物碱的含量。 方法 采用索式提取法对药材进行提取,结合酸溶碱沉和萃取的方法从药材中提取白喉乌头总生物碱;利用紫外光谱对经过酸性染料络合的生物碱进行分析,检测其中总生物碱的含量。 结果 新源县、巩留县和昭苏县所产白喉乌头原药材中含有总生物碱分别为2.246,4.682和3.597 mg·g^-1;回归方程为Y=11.5X + 0.121,r=0.999 6,乌头碱浓度在6～36 μg·mL^-1内呈良好的线性关系。 结论 采用紫外-可见分光光度法测定不同产地白喉乌头中总生物碱的含量,具有操作简便、结果准确的特点;其结果可为该药材的活性和毒性物质基础研究提供依据。     

银杏叶总黄酮对小鼠脑缺血再灌注损伤的保护作用

目的:建立拟脑梗死模型,观察银杏叶总黄酮预处理对小鼠脑梗死的保护作用并分析其作用机制。方法:脑缺血再灌注方法制备拟脑梗死小鼠模型,将小鼠随机分为对照组、模型组、用药组(高、中、低3个剂量组)。用药组腹腔注射银杏叶总黄酮注射液,模型组和对照组在同一时间给予腹腔注射等量生理盐水。末次给药2h后进行脑缺血再灌注手术。最后测定血清超氧化物歧化酶(SOD)、丙二醛(MDA)的含量;取脑组织做苏木素-伊红染色。结果:脑缺血再灌注可造成小鼠脑梗死,同时使血清中SOD活性降低,MDA含量升高。给予银杏叶总黄酮预处理后,可提高血清中SOD活性,降低MDA含量。结论:银杏叶总黄酮对小鼠脑梗死具有一定的保护作用,其作用机制可能与银杏叶总黄酮抗氧化作用有关。     

评价HPPH对细胞色素P450酶体外代谢活性的影响

目的:评价HPPH在体外对大鼠及人肝微粒CYP450酶的6种亚型酶活性的影响,预测使用HPPH可能出现的药物相互作用。方法:将注射用HPPH与CYP450酶的6种亚型的特异性探针底物非那西汀(CYP1A2)、甲苯磺丁脲(CYP2C9)、S-美芬妥因(CYP2C19)、右美沙芬(CYP2D6)、氯唑沙宗(CYP2E1)、咪达唑仑(CYP3A4)和睾酮(CYP3A4)与大鼠及人肝微粒进行孵育反应,采用HPLC-MS/MS法测定对应的7种代谢产物(对乙酰氨基酚、羟基甲苯磺丁脲、4-羟基美芬妥因、O-去甲基右美沙芬、6-羟基氯唑沙宗、1′-羟基咪达唑仑、6β-羟基睾酮)的浓度。结果:在本实验条件下,HPPH浓度为1.00~50.00μmol·L-1时,未发现其对大鼠的CYP1A2、CYP2C9、CYP2D6、CYP2E1、CYP3A4产生抑制作用。在本实验条件下,HPPH浓度为0.50~10.00μmol·L-1时,未发现其对人CYP1A2产生抑制作用;但对人CYP2C9、CYP2C19、CYP2D6、CYP2E1均存在抑制作用;对人CYP3A4,对底物咪达唑仑存在抑制作用,对底物睾酮未发现抑制作用。结论:HPPH对CYP450酶的抑制作用存在种属差异,在人体内HPPH与CYP450酶作用有待体内实验进一步证明。     

Stress Degradation Behavior of Abacavir Sulfate and Development of a Suitable Stability-Indicating UHPLC Method for the Determination of Abacavir,its Related Substances,and Degradation Products

A novel, stability-indicating UHPLC method was developed for the quantitative determination of Abacavir sulfate, its related substances, and forced degradation impurities in bulk drugs. The chromatographic separation was achieved on a Waters Acquity BEH C₈, 50 mm × 2.1 mm, 1.7 μm particle size column with a mobile containing a gradient mixture of solution A (0.10 % v/v o-phosphoric acid in water) and solution B (0.10% v/v o-phosphoric acid in methanol). The flow rate was set at 0.40 mL/min and the run time was 6.0 min. The drug substance was subjected to the stress studies of hydrolysis, oxidation, photolysis, and thermal degradation. Abacavir sulfate was found to degrade significantly under acidic hydrolysis and oxidative stress conditions. The formed degradation products were reported and were well-resolved from Abacavir and its related substances. The mass balance was found to be satisfactory in all of the stress conditions, thus proving the stability-indicating capability of the method. The developed UHPLC method was validated to be in agreement with ICH requirements and found to be rapid, accurate, precise, linear, specific, and suitable for the quantitative determination of related substances and degradants in the bulk drug samples of Abacavir sulfate.     

Effect of cooking on the concentration of Vitamins B in fortified meat products

B vitamins fortification of meat products is useful to compensate the loss of these compounds occurring during the heat treatment. The objective of this study was to evaluate the influence of heat treatments on the B vitamins concentration in fortified meat products. A rapid and reliable method for the simultaneous determination of Vitamins B₁, B₆ and B₁₂ in homogenized boiled ham and in various fortified burgers was set up. Extraction procedure and HPLC method ensure low detection limits, good sensitivity and resolution. Results showed that cooking processes caused a decrease in the B vitamins content both in mild (70–90 °C) and severe (120 °C) conditions. Performing a fortification of 25 μg g⁻¹ the residual concentration of B vitamins after cooking allow to reach the recommended daily allowance, thus suggesting that B vitamins fortification of meat product is an useful practice.     

Stress Degradation Behavior of Atorvastatin Calcium and Development of a Suitable Stability-Indicating LC Method for the Determination of Atorvastatin,its Related Impurities,and its Degradation Products

A rapid, reversed-phase liquid chromatographic method was developed for the quantitative determination of Atorvastatin calcium, its related substances (12 impurities), and degradation impurities in bulk drugs. The chromatographic separation was achieved on a Zorbax Bonus-RP column by employing a gradient elution with water–acetonitrile–trifluoroacetic acid as the mobile phase in a shorter run time of 25 min. The flow rate was 1.0 mL/min and the detection wavelength was 245 nm. The drug substance was subjected to stress studies such as hydrolysis, oxidation, photolysis, and thermal degradation, and considerable degradation was observed in acidic hydrolysis, oxidative, thermal, and photolytic stress conditions. The formed degradation products were reported and were well-resolved from the Atorvastatin and its related substances. The stressed samples were quantified against a qualified reference standard and the mass balance was found to be close to 99.5% (w/w) when the response of the degradant was considered to be equal to the analyte (i.e. Atorvastatin), which demonstrates the stability-indicating capability of the method. The method was validated in agreement with ICH requirements. The method developed here was single and shorter (25 min method for the determination of all 12 related impurities of Atorvastatin and its degradation products), with clearly better resolution and higher sensitivity than the European (85 min method for the determination of six impurities) and United States pharmacopeia (115 min and 55 min, two different methods for the determination of six related substances).