大黄和虎杖对血瘀症模型大鼠血流动力学和血脂的影响

目的观察大黄和虎杖及其配伍对血瘀症模型大鼠血流动力学和血脂的影响。方法将40只SPF级雌性大鼠随机分为空白组、模型组、实验Ⅰ组、实验Ⅱ组和实验Ⅲ组,每组8只。实验Ⅰ、Ⅱ、Ⅲ组分别予以30 mg·kg^-1大黄、虎杖、大黄虎杖(1∶1),qd;空白组和模型组均按5 mL·kg^-1的剂量予以0.9%NaCl,qd。5组大鼠均连续灌胃10 d。第10天灌胃1 h后,除空白组外,其余各组均予以盐酸肾上腺素1 mg·kg^-1,腹腔注射。24 h后,再次注射相同剂量的肾上腺素。急性血瘀模型制备1 h,将大鼠麻醉后腹主动脉采血。比较4组大鼠的血流动力学和血脂指标。结果空白组和实验Ⅲ组的全血黏度200/s分别为(3.26±1.33)和(4.51±1.15)mPa·s,50/s分别为(4.21±1.41)和(5.80±1.44)mPa·s,5/s分别为(9.75±2.97)和(13.27±3.14)mPa·s,1/s分别为(25.58±7.24)和(34.49±7.95)mPa·s,差异均有统计学意义(均P<0.05)。空白组、实验Ⅰ组、实验Ⅱ组红细胞聚集指数分别为(8.04±0.80),(7.39±0.31)和(7.18±0.62),空白组与实验Ⅰ组比较差异有统计学意义(P<0.05),空白组与实验Ⅱ组比较差异有统计学意义(P<0.01)。模型组和实验Ⅱ组的三酰甘油分别为(0.92±0.39)和(0.61±0.15)mmol·L^-1,差异有统计学意义(P<0.05)。结论大黄、虎杖单用和大黄虎杖配伍合用均对血瘀证大鼠的血流动力学异常有较好的改善作用,但大黄虎杖1∶1配伍没有表现出明显的协同作用。虎杖能显著降低三酰甘油,而大黄单用没有表现明显降低血脂的作用,在降低血脂方面大黄和虎杖配伍合用也没有表现出协同作用。     

Membrane permeability-guided identification of neuroprotective components from Polygonum cuspidatun

Context: Polygonum cuspidatum Sieb et Zucc. (Polygonaceae) possesses various pharmacological activities and has been widely using as one of the most popular and valuable Chinese herbal medicines in clinics. Its usage has increasingly attracted much of our attention and urges investigation on its bioactive components.

Objective: To establish a rapid and valid approach for screening potential neuroprotective components from P. cuspidatum.

Materials and methods: Potential neuroprotective components from P. cuspidatum were screened utilizing liposome equilibrium dialysis followed by high-performance liquid chromatography (HPLC) analysis. Their neuroprotective effects on modulation of protein expression of α7 nAChR, α3 nAChR and synaptophysin (SPY) on SH-SY5Y human neuroblastoma cell line (SH-SY5Y) were evaluated by means of Western blotting.

Results: Two potential compounds, polydatin (C1) and emodin-8-O-β-d-glucoside (C2), were detected and identified in our study. The biological tests showed that both compounds C1 and C2, respectively, at concentrations of 0.1 and 0.25?mg/mL significantly increased protein expression of α7 and α3 nicotinic acetylcholine receptors (nAChRs) in SH-SY5Y cells. Moreover, C1 and C2 at 0.1?mg/mL significantly reversed the Aβ_1-42-induced decrease of α7 and α3 nAChRs protein expression in SH-SY5Y cells. In addition, C2 at 0.1?mg/mL significantly increased protein expression of SPY in SH-SY5Y cells and Aβ_1-42-induced SH-SY5Y cells whereas C1 did not provide any positive effects.

Discussion and conclusion: In conclusion, our approach utilizing liposome equilibrium dialysis combined with HPLC analysis and cell-based assays is a prompt and useful method for screening neuroprotective agents.     

虎杖药材的HPLC-DAD-ELSD指纹图谱研究

目的建立虎杖药材的HPLC-DAD-ELSD指纹图谱,对10个不同产地的虎杖药材进行质量评价。方法使用Luna C_(18)(2)色谱柱(250mm×4.6mm,5μm),乙腈-水为流动相进行梯度洗脱,体积流量1.0mL/min,柱温35℃,检测波长254 nm,进样量10μL。蒸发光检测器漂移管温度109℃,载气体积流量3.0 L/min。结果通过10批虎杖药材建立了指纹图谱的共有模式,其中HPLC-DAD指纹图谱共有峰19个,相似度为0.938~0.993;HPLC-ELSD指纹图谱共有峰14个,相似度为0.905~0.999。确认了虎杖苷、白藜芦醇、大黄素-8-O-β-D-葡萄糖苷、大黄素甲醚-8-O-β-D-葡萄糖苷、大黄素和大黄素甲醚。结论方法准确、稳定,为客观评价虎杖药材的质量提供了科学依据。     

Effects of Ixeris Chinensis (Thunb.) Nakai Boiling Water Extract on Hepatitis B Viral Activity and Hepatocellular Carcinoma

Background

Hepatitis B virus (HBV) infection and hepatocellular carcinoma are major diseases that affect the Taiwanese population. Therefore, the development of an alternative herbal medicine that can effectively treat these diseases is a research target. In this study, we tested Ixeris Chinensis (Thunb.) Nakai boiling water extract (ICTN BWE) in vitro and analysed its effects on the HBV and liver cancer.

Materials and Methods

We used a human liver cancer cell line (Hep3B, a cell line that continuously secretes HBV particles into a medium) as an experimental model for the screening of various ICTN BWE concentrations and their effects on the HBV in vitro.

Results

Our results showed that 75 µg/mL ICTN BWE downregulated the relative expression of the hepatitis B virus surface antigens (HBsAg) to 77.1%. Using the human liver cancer cell lines HuH-7 and HepG2, and 3-(4,5-dimethylthiazol-zyl)-2,5-diphenyl tetrazolium bromide (MTT) and tumour clonogenic assays, we then showed that ICTN BWE inhibits hepatocellular carcinoma growth.

Conclusion

Fluorescent microscopy of DAPI(4′,6-Diamidino-2-phenylindole)-stained nuclei and DNA fragmentation assays confirmed the inhibitory effects of ICTN BWE on liver tumour cell growth through induction of apoptosis.     

Effects of Polysaccharides from Gynostemma Pentaphyllum (Thunb.), Makino on Physical Fatigue

Background

Gynostemma pentaphyllum (Thunb.) Makino has been reported to have a wide range of health benefits in Chinese herbal medicines. Polysaccharides from Gynostemma pentaphyllum (PGP), has been identified as one of the active ingredients responsible for its biological activities. Although many pharmacological activities of PGP have received a great deal of attention, there is limited evidence for the anti-fatigue effects of PGP. The purpose of this study was to investigate the effects of polysaccharides from PGP on physical fatigue.

Materials and method

The rats were divided into four groups, with 10 animals per group: control (C), group, low-treated (LT), group, medium-treated (MT), group, and high-treated (HT), group. The C group received distilled water, while LT, MT and HT groups were given various doses of PGP (100, 200, 400 mg/kg· d). After 30 days, forced swimming test was carried out in an acrylic plastic pool, then the exhaustive swimming time of rats and some biochemical parameters related to fatigue were measured. The data obtained showed that PGP could extend the exhaustive swimming time of the rats, as well as decrease the blood lactic acid (BLA), and blood urea nitrogen (BUN), concentrations, and increase the hemoglobin, liver glycogen and muscle glycogen concentrations.

Result

The data obtained showed that different doses of PGP could extend the exhaustive swimming time of the rats, as well as decrease the BLA and BUN concentrations, and increase the hemoglobin, liver glycogen and muscle glycogen concentrations, which suggests that PGP had significant anti-fatigue effects on rats.

Conclusion

PGP may be of use as a potential anti-fatigue agent, but there is a need for further research on long-term use in order to show its positive effects on physical fatigue.     

水栀子及京尼平苷抑制RBL-2H3细胞脱颗粒作用的代谢组学研究

目的 基于细胞代谢组学的方法研究水栀子Gardenia jasminoside及其主要有效成分京尼平苷对RBL-2H3细胞脱颗粒模型的干预作用。方法 利用UPLC-QTOF-MS高分辨质谱检测RBL-2H3细胞的代谢轮廓改变,并应用主成分分析（PCA）和正交偏最小二乘判别分析（OPLS-DA）筛选出引起这种差异的代谢标志物;同时应用MEV软件对差异标志物水平的变化进行聚类分析和热图绘制。结果 分别得到与水栀子和京尼平苷作用相关的代谢标志物54和46个,其中共有标志物31个,且在2个给药组中变化趋势相同。由共有代谢标志物富集得到了受干扰的5条代谢通路：甘氨酸、天冬氨酸、谷氨酸代谢,谷胱甘肽代谢,组胺代谢,能量代谢,烟酰胺代谢。结论 水栀子和京尼平苷通过调节组胺代谢、氧化应激和能量代谢而抑制RBL-2H3细胞脱颗粒过程,且京尼平苷是水栀子发挥作用的主要药效物质基础之一。     

虎杖MYB转录因子PcMYB1的表达特性和功能研究

目的对虎杖中1个新的R2R3-MYB转录因子基因PcMYB1进行转录活性鉴定和表达特性分析,并在转基因拟南芥中进行功能研究。方法利用酵母单杂交实验分析PcMYB1的转录活性;荧光定量PCR(RT-PCR)技术检测虎杖中PcMYB1的表达模式;利用Wiesner染色和溴乙酰法检测PcMYB1转基因拟南芥中的木质素含量;RT-PCR技术分析PcMYB1转基因拟南芥木质素合成相关基因的表达。结果酵母单杂实验结果表明,PcMYB1具有转录抑制活性;RT-PCR结果显示PcMYB1在虎杖根、茎、叶中均有表达,在叶中表达量最高,并且紫外照射处理可诱导叶片中PcMYB1的表达;与野生型拟南芥相比,转基因拟南芥的株高降低了24.07%,木质部的细胞染色程度浅,转基因拟南芥木质素含量降低了14.81%,参与木质素合成的AtC4H、AtC3H、AtF5H、AtCOMT、AtCAD基因表达下调。结论 PcMYB1具有转录抑制活性,对植物木质素合成具有负调控作用。     

HPLC法测定草珊瑚中富马酸含量的方法学研究

目的:建立高效液相色谱测定草珊瑚中富马酸含量的方法。方法:采用高效液相色谱法,固定相为Dia-monsil C18柱(5μm,250×4.6mm),以乙腈-水-磷酸(2:98:0.05)作为流动相,检测波长为208nm。结果:富马酸在54.35~434.8μg范围内线性关系良好,r2=0.9999,平均回收率为99.95%,RSD=1.33%。结论:该方法操作简单、准确,重复性好,可适用于草珊瑚药材的质量控制。     

何首乌水提物对光化学反应诱导的大鼠肠系膜细静脉血栓形成的抑制作用

目的:探讨何首乌水提物对光化学反应诱导的大鼠肠系膜细静脉血栓形成的抑制作用。方法:将24只雄性Wistar大鼠随机分为正常组、模型组、何首乌组和阿司匹林组,每组6只。用光化学反应法诱导肠系膜细静脉血栓形成,观察各组血栓出现时间、血栓占血管面积一半所需时间、诱导停止后5min、10min、20min和30min时,各组血栓/血管面积比;然后在各组大鼠肠系膜表面滴加甲苯胺蓝,观察细静脉周围肥大细胞脱颗粒,并计算脱颗粒的肥大细胞比率。结果:何首乌水提物可以明显延迟血栓出现时间,明显缩短血栓占血管面积一半所需时间,显著降低血栓/血管面积比,显著减少血管周围肥大细胞脱颗粒。结论:何首乌水提物能明显抑制光化学反应诱导的大鼠肠系膜细静脉血栓形成。该作用可能与其抗氧化、抑制肥大细胞脱颗粒等密切相关。     

头花蓼总黄酮提取工艺研究

目的 探讨头花蓼总黄酮最佳提取工艺.方法 采用正交实验方法 进行优选,紫外分光光度法测定总黄酮含量.结果 乙醇浓度对总黄酮的提取有显著影响.结论 结合正交实验结果 和方差分析,确定12倍量55%乙醇提取3次,1h/次为最佳工艺.