Dammarane-type saponins from steamed leaves of Panax notoginseng

Four new dammarane-type triterpenoid saponins,namely notoginsenosides SFt1-SFt4(1-4)were isolated from the steamed leaves of Panax notoginseng(Burk.)F.H.Chen(Araliaceae),together with 17 known saponins.Their structures were established on the basis of detailed spectroscopic analyses and acidic hydrolysis.The known ginsenosides Rk2 and Rh3 were obtained from P.notoginseng for the first time.All of these new saponins showed no in vitro cytotoxicity against five human cancer cell lines(HL-60,SMMC-7712,A-549,MCF-7,and SW480).

Panax notoginseng saponins influence on transplantation of neural stem cell-derived dopaminergic neurons in a rat model of Parkinson’s disease

BACKGROUND： Dopaminergic neurons differentiated from neural stem cells have been successfully used in the treatment of rat models of Parkinson＇s disease; however, the survival rate of transplanted cells has been low. Most cells die by apoptosis as a result of overloaded intracellular calcium and the formation of oxygen free radicals. OBJECTIVE： To observe whether survival of transplanted cells, transplantation efficacy, and dopaminergic differentiation from neural stem cells is altered by Panax notoginseng saponins （PNS） in a rat model of Parkinson＇s disease. DESIGN, TIME AND SETTING： Cellular and molecular biology experiments with randomized group design. The experiment was performed at the Animal Experimental Center, First Hospital of Sun Yat-sen University from April to October 2007. MATERIALS： Thirty-two adult, healthy, male Sprague Dawley rats, and four healthy Sprague Dawley rat embryos at gestational days 14-15 were selected. The right ventral mesencephalon was injected with 6-hydroxydopamine to establish a model of Parkinson＇s disease. 6-hydroxydopamine and apomorphine were purchased from Sigma, USA. METHODS： Neural stem cells derived from the mesencephalon of embryonic rats were cultivated and passaged in serum-free culture medium. Lesioned animals were randomly divided into four groups （n = 8）： dopaminergic neuron, dopaminergic neuron ＋ PNS, PNS, and control. The dopaminergic neuron group was injected with 3 μL cell suspension containing dopaminergic neurons differentiated from neural stem cells. The dopaminergic neurons ＋ PNS group received 3 μ L dopaminergic cell suspension combined with PNS （250 mg/L）. The PNS group received 3 μL PNS （250 mg/L）, and the control group received 3 μL DMEM/F12 culture medium. MAIN OUTCOME MEASURES： The rats were transcardially perfused with 4% paraformaldehyde at 60 days post-grafting for immunohistochemistry. The rats were intraperitoneally injected with apomorphine （0.5 mg/kg） to induce rotational behavior. RESULTS： Cell counts of tyrosine hydroxylase-positive neurons in the dopaminergic neuron ＋ PNS group were （732±82.6） cells/400-fold field. This was significantly greater than the dopaminergic neuron group [（326 ± 34.8） cells/400-fold field, P 〈 0.01]. Compared to the control group, the rotational asymmetry of rats that received dopaminergic neuron transplants was significantly decreased, beginning at 20 days after operation （P 〈 0.01）. Rotational asymmetry was further reduced between 10-60 days post-surgery in the dopaminergic neuron ＋ PNS group, compared to the dopaminergic neuron group （P 〈 0.01）. CONCLUSION： Panax notoginseng saponins can increase survival and effectiveness of dopaminergic neurons differentiated from neural stem cells for transplantation in a rat model of Parkinson＇s disease.

Ginsenoside-Rg_6, a Novel Triterpenoid Saponin from the Stem-Leaves of Panax ginseng C. A. Mey.

A novel dammarane-type triterpene oligoglycoside, named ginsenoside-Rg6 3, was isolated from the stem-leaves of Panax ginseng C. A. Mey., together with two known ones, 20(S)-ginsenoside-Rg2 1 and 20(R)-ginsenoside-Rg2 2. On the basis of chemical and physicochemical evidence , the structure of ginsenoside-Rg6 have been elucidated as 6-O-(-L-rhamnosyl-(1?2)-(-D-glucopyranosyl-dammarane-(E)-20(22), 24-diene-3(, 6(, 12(-triol.

三七茎叶提取物对断奶仔猪生长性能、血清生化指标和肠道健康的影响

试验旨在研究三七茎叶提取物对断奶仔猪生长性能、血清生化指标和肠道健康的影响。选取26日龄断奶仔猪24头随机分为3组,每组8个重复,每个重复1头仔猪。对照组饲喂基础饲粮,低剂量组、高剂量组分别在基础饲粮中添加0.2%和0.4%的三七茎叶提取物。试验期21 d。结果显示,与对照组相比,高剂量组断奶仔猪平均日增重显著升高(P<0.05),低剂量组和高剂量组血清乳酸脱氢酶活性、谷草转氨酶活性和谷草/谷丙的比值显著降低(P<0.05);高剂量组断奶仔猪十二指肠、空肠、回肠的绒毛高度显著增加(P<0.05),十二指肠和回肠的绒隐比显著升高(P<0.05);低剂量组和高剂量组结肠白细胞介素-12 p35(IL-12 p35)、白细胞介素-12 p40(IL-12 p40)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和干扰素-γ(IFN-γ)的mRNA表达量显著降低(P<0.05)。研究表明,在饲粮中添加三七茎叶提取物可以改善断奶仔猪生长性能、肠道健康,添加0.4%三七茎叶提取物较为适宜。

基于网络药理学探究三七叶对糖尿病大鼠的治疗机制

目的 利用网络药理学和动物实验,研究三七叶抗糖尿病的主要活性成分和潜在作用机制。方法 基于Pubchem、PharmMapper、OMIM等数据库筛选三七叶与糖尿病共同的潜在靶点之后,再利用STRING数据库和Cytoscape软件构建蛋白相互作用网络图,对其进行拓扑学分析和聚类分析获得核心靶点,并进行GO和KEGG富集分析筛选药物作用于疾病的可能机制。随后进行动物实验验证三七叶的抗糖尿病作用。结果 预测结果共获得三七叶与糖尿病的共同靶点50个,核心靶点有3个:ANXA5、CFB、GSTP1,共涉及生物过程986种,细胞组成15种,分子功能74种,信号通路101条。动物实验表明三七叶可降低糖尿病大鼠的血糖,减少与糖尿病肾病相关的p38 MAPK和TGF-β1 mRNA的表达,同时也可以调节氧化应激和血脂水平,减少动脉粥样硬化。结论 三七叶是通过多途径多靶点共同发挥抗糖尿病的作用,其作用机制可能与减少氧化应激、调节细胞因子、减少动脉粥样硬化和抗糖尿病肾病肾纤维化等有关。

UPLC-MS/MS测定心悦胶囊中6个皂苷类成分的含量

目的:采用超高效液相色谱-质谱法(UPLC-MS/MS)建立心悦胶囊中主要成分人参皂苷Rg_(1)、人参皂苷Re、人参皂苷Rb_(1)、拟人参皂苷F_(11)、人参皂苷Rd和20(S)-人参皂苷F1的含量测定方法。方法:使用Waters ACQUITY BEH C_(18)色谱柱(100 mm×2.1 mm,1.7μm),以乙腈-水为流动相,梯度洗脱,流速为0.5 mL·min^(–1),柱温为40℃,电喷雾电离源,多反应离子监测扫描方式进行检测。结果:6个成分分别在质量浓度为9.638~963.800、9.839~983.900、9.951~995.100、5.270~1054.000、9.608~960.800、4.905~981.000 ng·mL^(–1)时与峰面积线性关系良好(r>0.998),平均加样回收率(RSD)分别为96.9%(2.2%)、100.1%(1.5%)、97.2%(1.8%)、98.5%(2.3%)、96.6%(2.8%)、100.2%(3.5%)。结论:所建立的方法快速、准确、灵敏度高,可用于心悦胶囊中西洋参茎叶总皂苷的质量控制。

Chinese preparation Xuesaitong promotes the mobilization of bone marrow mesenchymal stem cells in rats with cerebral infarction

After cerebral ischemia, bone marrow mesenchymal stem cells are mobilized and travel from the bone marrow through peripheral circulation to the focal point of ischemia to initiate tissue regeneration. However, the number of bone marrow mesenchymal stem cells mobilized into peripheral circulation is not enough to exert therapeutic effects, and the method by which blood circulation is promoted to remove blood stasis influences stem cell homing. The main ingredient of Xuesaitong capsules is Panax notoginseng saponins, and Xuesaitong is one of the main drugs used for promoting blood circulation and removing blood stasis. We established rat models of cerebral infarction by occlusion of the middle cerebral artery and then intragastrically administered Xuesaitong capsules（20, 40 and 60 mg/kg per day） for 28 successive days. Enzyme-linked immunosorbent assay showed that in rats with cerebral infarction, middle- and high-dose Xuesaitong significantly increased the level of stem cell factors and the number of CD117-positive cells in plasma and bone marrow and significantly decreased the number of CD54-and CD106-positive cells in plasma and bone marrow. The effect of low-dose Xuesaitong on these factors was not obvious. These findings demonstrate that middle- and high-dose Xuesaitong and hence Panax notoginseng saponins promote and increase the level and mobilization of bone marrow mesenchymal stem cells in peripheral blood.

Transformation of Compound K from Saponins in Leaves of Panax notoginseng by Immobilized β-Glucanase

Objective To prepare an active anti-tumor component,compound K(C-K),from saponins in leaves of Panax notoginseng(SLPN) using immobilized β-glucanase.Methods Two entrapments,alginate gel-1(Alg 1) and alginate gel-2(Alg 2),were evaluated for their ability to immobilize β-glucanase.The amount and purity of C-K obtained from the transformation process were analyzed by HPLC,and the immobilizing parameters were optimized.Results β-Glucanase can be immobilized and reused with either of the entrapment.However,using Alg 1 resulted in higher enzyme activity than Alg 2.The optimal concentration of the immobilized enzyme was 10%;The optimal crosslinking time was 4–6 h;and the optimal concentration of the crosslinking agent was 6%– 7%.Conclusion Immobilized β-glucanase shows sustained enzyme activity,good ethanol tolerance,and was reusable for the preparation of C-K from SLPN.

4种真菌发酵对三七叶化学成分及药理活性的影响

以三七叶为原料,选择米根霉、粗糙脉孢菌、红曲霉和鲁氏毛霉4种真菌对其进行发酵,测定发酵过程中三七叶总皂苷、总多糖、总酚和总黄酮的含量变化,评估其体外抗氧化活性和α-葡萄糖苷酶抑制活性的变化,并采用超高效液相色谱-质谱对发酵前后三七叶的代谢谱进行分析。结果表明:4种真菌均可使三七叶中总皂苷和总多糖含量显著降低,但米根霉、粗糙脉孢菌和红曲霉可提高总酚含量,而米根霉和鲁氏毛霉可提高总黄酮含量。真菌发酵3~5 d后,三七叶的抗氧化活性明显增强,其中红曲霉发酵3 d的增强效果最好,可使三七叶的1,1-二苯基-2-三硝基苯肼自由基、2,2’-联氮双(3-乙基苯并噻唑啉-6-磺酸)阳离子自由基清除能力以及铁离子还原能力分别提高14.30%、5.13%、18.40%。此外,真菌发酵可先降低后提高三七叶的α-葡萄糖苷酶抑制活性,其中红曲霉在发酵6 d后可使其提高16.03%。进一步的非靶向代谢组学分析,从三七叶中共鉴定出573种代谢物,经红曲霉发酵3 d后,其上调代谢物明显少于下调代谢物,主要为氨基酸、生物碱、碳水化合物、脂类、酚类、类黄酮以及萜类化合物等;对其中的21种皂苷类化合物进行进一步分析,发现其中只有人参皂苷F2显著上调,另有8种皂苷显著下调;对差异代谢物进行京都基因与基因组百科全书富集分析发现,嘌呤代谢、嘧啶代谢、类黄酮生物合成、辅助因子的生物合成等是三七叶在发酵过程中最可能的代谢通路。综上,真菌发酵尤其是红曲霉发酵是改善三七功能活性的绿色有效手段,其可通过多个作用途径使三七叶的代谢物变化,影响其功能活性,本研究可为三七叶的绿色加工提供科学参考。

三七茎秆离散元参数标定与试验

针对利用离散元法进行三七联合收获、茎秆杀秧等关键作业过程仿真分析时,三七茎秆本征参数、三七茎秆及作业装备间接触参数缺乏问题,以三七茎秆为对象,利用EDEM软件建立三七茎秆离散元Hertz-Mindlin/Hertz-Mindlin with bonding模型,通过堆积角试验和虚拟仿真试验对三七茎秆离散元参数进行标定,并建立三七茎秆杀秧装置模型。通过力学特性试验确定三七茎秆本征参数;采用圆筒提升法进行三七茎秆堆积角试验,使用Origin软件对堆积角图像进行轮廓拟合得到三七茎秆堆积角为44.53°;设计Placktt-Burman试验、最陡爬坡试验和Central-Composite试验确定三七茎秆及作业装备间接触参数,并利用堆积角试验和剪切试验验证模型的可靠性。结果表明:三七茎秆与作业装备间碰撞恢复系数、静摩擦因数、滚动摩擦因数分别为0.319、0.25、0.029;三七茎秆间的碰撞恢复系数、静摩擦因数、滚动摩擦因数最优值分别为0.4、0.29、0.032;Hertz-Mindlin with bonding模型法向刚度Kn为3.26×10^(8)N/m^(3)、切向刚度Ks为2.17×10^(8)N/m^(3)、法向临界应力σ为2.27 MPa、切向临界应力γ为9.65 MPa、粘结半径Rd为0.1 mm;堆积角验证试验中相对误差为0.29%;剪切验证试验中相对误差为1.52%,误差较小。三七茎秆离散元模型与实际情况基本一致,三七茎秆模型和标定离散元仿真参数可靠,可为三七茎秆离散元仿真研究提供参考。

三七茎叶-滇黄精速溶粉提取工艺及抗氧化活性分析

为探索三七茎叶-滇黄精复合产品的开发工艺,该研究分析了三七茎叶-滇黄精速溶粉中可溶性总糖、皂苷、多酚化合物的组成及含量,并测定其抗氧化活性。以单因素-响应面实验考察三七茎叶-滇黄精在乙醇添加量、料液比(g∶mL)、浸提时间3种变量条件下的最佳浸提条件,进一步以喷雾干燥的方法获取三七茎叶-滇黄精速溶粉,以蒽酮比色法测定速溶粉中可溶性总糖的含量,高效液相色谱法测定速溶粉中皂苷、多酚含量,进一步测定其抗氧化活性,并进行感官审评。喷雾干燥优化出三七茎叶与滇黄精的最佳质量比为6∶4,最佳浸提工艺为三七茎叶-滇黄精的料液比(g∶mL)为1∶21,29%(体积分数)乙醇,浸提37 h,重复浸提2次,每1 kg复合物可得到速溶粉374.23 g,得粉率37.42%,该速溶粉溶解性好,香甜回甘。速溶粉中可溶性总糖含量为(55.44±3.21)mg/g,活性成分以三七皂苷Fa、Fd、Fc、人参皂苷Rb1等7种皂苷及咖啡酸(caffeic acid,CA)、表儿茶素没食子酸酯(epicatechin-3-gallate,ECG)、表没食子儿茶素没食子酸酯(epigallocahechin-3-gallate,EGCG)等5种酚类化合物为主。抗氧化活性结果表明,三七茎叶-滇黄精速溶粉对ABTS阳离子自由基、DPPH自由基、超氧阴离子自由基、羟自由基具有较好的清除能力,其IC 50值分别为0.30、0.63、0.49、1.67 mg/mL。该研究优化了三七茎叶-滇黄精浸提工艺,所得速溶粉富含皂苷、多酚、可溶性糖等成分,为三七茎叶和滇黄精在食品开发领域提供基础研究数据,为云南特色食品及药食同源产品的开发利用提供科学支撑。

凝结芽胞杆菌发酵三七叶茶抗氧化活性及其香气成分研究

为探究凝结芽孢杆菌接种量对三七叶茶功效成分、抗氧化活性和香气成分的影响,设置了4种凝结芽胞杆菌接种量(0%、1%、3%和5%)发酵三七叶茶,分析了发酵三七叶茶的总酚、总黄酮、总皂苷、DPPH自由基和ABTS阳离子自由基清除率、挥发性香气成分和感官评价。结果表明,凝结芽孢杆菌发酵三七叶茶总酚含量和总黄酮含量显著降低了6.8%~21.3%和28.4%~40.9%,且随接种量增加,发酵三七叶茶的总酚含量和总黄酮含量均呈先增大后减小的趋势;接种凝结芽孢杆菌使发酵三七叶茶的总皂苷含量显著增加了0.085~1.45倍,醇提物DPPH自由基、ABTS阳离子自由基清除率显著增加了12%~28.8%、9.6%~22.5%,且挥发性成分种类和浓度均有增加。此外,凝结芽孢杆菌能提高三七叶茶的感官总评分。其中,3%凝结芽孢杆菌接种量发酵的三七叶茶总皂苷含量最高,为373.7 mg/g,且抗氧化效果和感官评价最好,推荐作为发酵三七叶茶的适宜接种量。该研究结果可为优化三七叶茶发酵工艺,实现三七叶片的高效利用提供理论依据和科学指导。

干燥方式对三七叶主要活性成分、体外抗氧化、α-葡萄糖苷酶抑制活性、挥发性成分和代谢物的影响

为选择较好的三七叶干燥方式,分别采用热风干燥、热泵干燥、真空干燥3种方式对三七叶进行干燥,测定不同干燥方式下三七叶的总皂苷、多糖、总酚和总黄酮含量以及单体皂苷含量,评估其体外抗氧化和α-葡萄糖苷酶抑制能力,采用气相色谱-离子迁移谱联用技术鉴定其挥发性成分,并采用非靶向代谢组学技术分析其代谢物变化。结果表明:3种干燥方式均可明显提高三七叶的总皂苷和多糖含量,降低其总酚和总黄酮含量,但均不同程度地减弱了其功能活性;其中热泵干燥组的总皂苷和多糖含量最高,并具有最强的1,1-二苯基-2-三硝基苯肼自由基清除能力和α-葡萄糖苷酶抑制能力,以及较强的2,2’-联氮-双-3-乙基苯并噻唑啉-6-磺酸阳离子自由基清除能力和铁还原能力;高效液相色谱分析发现热泵干燥显著降低了三七叶中三七皂苷Fc、Fe的含量,而真空干燥组与之相反;从三七叶中共检测出85种挥发性成分,其中热泵干燥组的醇类和醛类化合物相对含量最高,而酮类化合物相对含量最低,且不同处理组之间有明显区别;采用超高效液相色谱-四极杆-轨道阱质谱联用技术从热泵干燥前后的三七叶中鉴定出659种代谢物,其中有113种和68种代谢物分别在正、负离子模式下表达显著上调,98种和31种代谢物分别在正、负离子模式下表达显著下调,主要包括脂质和类脂分子、有机杂环化合物、有机酸及其衍生物等。针对三七叶中24种皂苷类代谢物进行进一步分析,发现其中7种相对含量显著降低、1种相对含量显著升高,这也印证了高效液相色谱的测定结果;对差异代谢物进行京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析,发现新鲜三七叶在热泵干燥过程中最可能存在的代谢通路有氨基酸的生物合成、2-氧羧酸代谢及辅助因子的生物合成。综上,热泵干燥对三七叶的化学成分和功能活性具有最好的保持效果,较适合用于三七叶的干燥,其可通过多个作用途径改变三七叶的代谢物组成,并影响其挥发性成分含量。

饲粮中添加三七茎叶对西畴乌骨鸡血清生化指标、免疫功能以及肌肉品质的影响

试验旨在研究在饲粮中添加三七茎叶对西畴乌骨鸡血清生化指标、免疫功能以及肌肉品质的影响。选择42日龄的西畴乌骨鸡270只随机分为3组,每组3个重复,每个重复30只。对照组乌骨鸡饲喂基础饲粮,Ⅰ组、Ⅱ组分别在基础饲粮中添加4%、8%三七茎叶。试验期90 d。结果显示,Ⅰ组、Ⅱ组乌骨鸡血清球蛋白含量、法氏囊指数、脾脏指数以及肌肉中苏氨酸、缬氨酸、异亮氨酸、苯丙氨酸、赖氨酸、精氨酸、棕榈油酸、油酸、γ-亚麻酸、二十碳五烯酸、二十二碳六烯酸以及必需氨基酸的含量显著提高(P<0.05),血清中碱性磷酸酶、尿素氮含量以及肌肉中豆蔻酸、硬脂酸含量显著降低(P<0.05)。研究表明,在饲粮中添加三七茎叶饲喂西畴乌骨鸡可改善其血清生化指标,调节机体代谢功能,提升免疫力,改善西畴乌骨鸡的肌肉品质,添加4%三七茎叶较为适宜。

西洋参茎叶提取物对鹌鹑生长性能和肌肉品质的影响

【目的】探讨西洋参茎叶提取物对鹌鹑生长性能、肌肉品质的影响,为西洋参茎叶作为添加剂开发利用提供依据。【方法】将150只1日龄鹌鹑,随机分成5组,每组3个重复,每个重复10只鹌鹑。空白对照组(CON)饲喂基础日粮,抗生素对照组(KSS)在基础日粮中添加阿莫西林(100 mg·kg^(-1)),3个试验组在基础日粮中分别添加1%(SD)、1.5%(SZ)和2%(SG)西洋参茎叶提取物。试验期50 d,预饲期5 d,正饲期45 d。【结果】从鹌鹑生长性能来看,SZ组可显著提高鹌鹑平均日增质量(P<0.05),同时显著降低料肉比(P<0.05)。从鹌鹑肉嫩度来看,与CON组和KSS组相比,试验组鹌鹑胸肌与腿肌剪切力显著降低(P<0.05),肉嫩度均增强,其中SZ组肉嫩度最佳;从鹌鹑肉中营养成分来看,鹌鹑胸肌中蛋白质含量SG组显著高于CON组(P<0.05),SZ和SG组脂肪含量显著高于CON组和KSS组(P<0.05);鹌鹑腿肌中蛋白质含量SZ和SG组显著高于CON组和KSS组(P<0.05),脂肪含量SZ组显著高于CON组和KSS组(P<0.05);胸肌和腿肌中水分含量各组间差异不显著(P>0.05)。从鹌鹑肌肉中脂肪酸含量来看,与CON组相比,各试验组鹌鹑胸肌中不饱和脂肪酸含量降低不显著(P>0.05),SZ组鹌鹑腿肌中不饱和脂肪酸含量显著升高(P<0.05);从鹌鹑肌肉中肌苷酸含量来看,SZ组鹌鹑胸肌和腿肌中肌苷酸含量均显著高于KSS组(P<0.05);从鹌鹑肌肉中氨基酸含量来看,SD、SZ和SG组鹌鹑胸肌和腿肌中必需氨基酸和风味氨基酸含量均显著高于KSS组(P<0.05)。从H.E.染色结果来看,各试验组鹌鹑的肝肾组织结构均未见异常。鹌鹑生长激素(Growth hormone,GH)基因(GH基因)表达水平各试验组均显著高于CON组(P<0.05),且SZ组鹌鹑生长激素(GH)基因表达量最高。【结论】西洋参茎叶提取物可加速鹌鹑生长,改善肌肉品质,增加肌肉风味,且对肝肾发育无不良影响,安全性比较高,可代替抗生素作为中草药饲料添加剂进一步开发利用。

Effects of Panax Notoginseng Saponins on Homing of C-kit^+ Bone Mesenchymal Stem Cells to the Infarction Heart in Rats

Objective:To investigate the effects of panax notoginseng saponins(PNS) on homing of C-kit+ bone mesenchymal stem cells(BMSCs) to the infarction heart.Methods:The acute myocardial infraction(AMI) model was established in 140 Wistar rats,105 model rats survived after operation,and the model rats were randomly divided into five groups,21 rats in each group:Western medicine group mobilized by subcutaneous injection of human granuloctye colony stimulating factor(G-CSF) 50 μg·kg-1·d-1;sham operation group and a model group treated by subcutaneous injection of normal saline 50 μg·kg-1·d-1;Chinese medicine group mobilized by intraperitoneal injection of Xuesaitong(血塞通)(ingredients of PNS) 150 mg·kg-1·d-1;integrative medicine group mobilized by subcutaneous injection of G-CSF 50 μg·kg-1·d-1 and intraperitoneal injection of Xuesaitong 150 mg·kg-1·d-1.Except for the sham-operated group,each group was divided into three sub-groups by three time points of 1 d,7 d and 14 d.G-CSF was injected once a day for 7 d.Xuesaitong was injected once a day until the rats were killed.The flow cytometry was used for detection of C-kit + cells in the peripheral blood in different time points,and immunohistochemical method was used for detection of the changes of C-kit + cell and Ki-67+ cell numbers in the marginal zone of AMI.Results:Twenty-four hours after the operation,C-kit + cells had a slight increase in the model group compared with the sham operation group(P>0.05).The peripheral blood C-kit+ cells in the integrative group increased significantly compared with the other groups on 7 d and 14 d(all P<0.05).Meanwhile the expression of C-kit + cells and Ki-67+ cells in the marginal zone of AMI in the integrative group increased significantly compared with the Chinese medicine group,the western medicine group and the model group on 1 d,7 d and 14 d(all P<0.05),and the cells in the integrative group decreased significantly on 14 d compared with that on 7 d(P<0.05).Conclusion:PNS can cooperate with G-CSF to mobilize C-kit+ BMSCs from the marrow into the peripheral blood and promote them "homing" to the infarction heart.

三七茎叶总皂苷抑制七氟醚诱导的海马神经元凋亡机制研究

目的探讨三七茎叶总皂苷(PNSLS)调节七氟醚诱导海马神经元凋亡的作用机制。方法以不同剂量七氟醚(0,4.1%,8%)干预小鼠海马神经元细胞系HT22,使用不同质量浓度的PNSLS(1.5,3.0,6.0,12.0μg/mL)预处理神经元。将HT22细胞分为对照组、暴露于4.1%七氟醚的细胞组(4.1%Sev组)、使用6μg/mL PNSLS对细胞进行预处理1 h后将细胞暴露于4.1%七氟醚组(4.1%Sev+PNSLS组)、在PNSLS处理前1 h加入丝裂原激活蛋白激酶(MAPK)激活剂anisomycin(An)再暴露于七氟醚和PNSLS的共培养组(4.1%Sev+PNSLS+An组)。通过CCK-8法检测神经元活力,流式细胞术及免疫印迹(Western blot)法检测海马神经元凋亡,比色活性测定试剂盒测定胱天蛋白酶-3(caspase-3)活性,Western blot法测定内质网应激相关蛋白及p38 MAPK/核转录因子-κB p65(p38 MAPK/NF-κB p65)信号通路相关蛋白的表达水平。结果4.1%七氟醚显著抑制海马神经元活力(P<0.05),6μg/mL PNSLS显著升高神经元活力并抑制海马神经元凋亡(P<0.05)。与Sev组比较,Sev+PNSLS组细胞中凋亡蛋白Bax表达水平显著降低(P<0.05),B细胞淋巴瘤2家族蛋白(Bcl-2)表达水平显著升高(P<0.05),caspase-3活性显著降低(P<0.05);Sev+PNSLS组内质网应激诱导的凋亡信号通路相关蛋白78000葡糖调节蛋白(GRP78)、内质网应激相关蛋白(CHOP)表达水平均显著降低(P<0.01),p38和IκBα的磷酸化水平均显著降低(P<0.01),细胞质中NF-κB p65水平显著升高(P<0.01),细胞核中NF-κB p65水平显著降低(P<0.01)。与Sev+PNSLS组比较,Sev+PNSLS+An组p38和IκBα的磷酸化水平均显著升高(P<0.01),细胞质中NF-κB p65水平显著降低(P<0.01),细胞核中NF-κB p65水平显著升高(P<0.01),海马神经元活力显著降低(P<0.01),凋亡率显著升高(P<0.01)。结论PNSLS通过激活p38 MAPK/NF-κB p65信号通路抑制七氟醚诱导的神经元凋亡。

消栓通络片中三七茎叶及重金属检查

目的建立消栓通络片中三七茎叶的检查方法,同时考察该制剂中重金属残留水平。方法采用薄层鉴别法对消栓通络片中三七茎叶进行筛查;运用电感耦合等离子体质谱法对样品中5种重金属有害元素残留量进行测定,并进行安全风险评估。结果置日光和紫外光(365 nm)下检视,样品色谱中在与三七茎叶皂苷对照提取物色谱相应的位置上未出现相同颜色的斑点;所有样品中有2批砷超标、1批汞超标,而安全风险评估结果显示该制剂整体上所含5种重金属的量对人体所构成的非致癌风险较低。结论所建薄层鉴别法可有效甄别消栓通络片中是否存在三七茎叶冒充三七投料现象,对重金属有害残留量的考察有助于控制该制剂的整体质量。

人参茎叶中人参皂甙含量检测前处理工艺优化及方法学考察

目的:优化人参茎叶中人参皂甙含量检测前处理的最佳工艺,确定人参茎叶原料中的人参皂甙含量检测方法并进行方法学考察。方法:采用正交实验优化人参茎叶中人参皂甙含量检测前处理工艺,采用高效液相色谱法测定人参总皂甙含量,并通过线性关系、精密度试验、重复性试验、稳定性试验、加样回收试验考察此方法的科学性与可行性。结果:原料前处理的最佳工艺为氧化镁加入量0.10 g,过40目筛,在50℃条件下超声提取1.5 h;方法精密度、重复性、稳定性良好,平均加样回收率为99.4%。结论:本文确立了人参茎叶原料的最优前处理方法,为人参皂甙含量的检测提供参考。

三七茎叶黄酮类成分的研究

目的对三七[Panax notoginseng(Burk.)F.H.Chen]茎叶的黄酮类成分进行分离鉴定。方法利用水煎煮提取、大孔吸附树脂吸附、硅胶柱色谱、ODS柱色谱等方法分离提纯,通过理化性质及IR,UV,MS,1H-NMR,13C-NMR等光谱数据鉴定其结构。结果分离得到6个黄酮类单体化合物,分别鉴定为:山柰酚(kaempferol,Ⅰ)、槲皮素(quercetin,Ⅱ)、山柰酚-7-O--αL-鼠李糖苷(kaempferol 7-O--αL-rhamnoside,Ⅲ)、山柰酚-3-O--βD-半乳糖苷(kaempferol 3-O--βD-galactoside,Ⅳ)、山柰酚-3-O--βD-半乳糖(2→1)葡萄糖苷[kaempferol 3-O-(2″--βD-glucopyranosyl)--βD-galactopyranoside,Ⅴ]、槲皮素-3-O--βD-半乳糖(2→1)葡萄糖苷[quercetin3-O-(2″--βD-glucopyranosyl)--βD-galactopyranoside,Ⅵ]。结论除山柰酚(kaempferol,Ⅰ)、槲皮素(quercetin,Ⅱ)之外,其余4个化合物皆为首次从该植物中分离得到,Ⅲ与Ⅵ为首次从该属植物中分离得到。