三七总皂苷及其主要成分对血管内皮细胞缺氧损伤的保护作用

研究三七总皂苷活血化瘀作用的血管内皮保护机制，并确定其血管的内皮保护作用的主要效应成分。方法：血管内皮细胞培养，MTT比色法测定药物毒性、台盼蓝染色、MTT比色法、LDH漏出率评价药效。结果：与模型组比较，三七总皂苷及其主要成分三七皂苷Rb、Rg1、Re组的LDH漏出率、细胞死亡率显著下降（P＜0．001），细胞存活率显著提高（P＜0．001）。结论：三七总皂苷的活血化瘀作用机制与其对血管内皮细胞缺氧的损伤的保护作用有关，三七皂苷Rb1、Rg1、Re是其血管内皮保护作用的主要效应成分。     

G-821树脂提取三七叶皂苷的工艺研究

三七为五加科植物三七Panax notoginseng(Burk)F.H.Chen的块根,具有活血化瘀、消肿止痛的作用,且能滋补强身。其主要活性成分为三七总皂苷。三七叶与块根有相似的药理作用,其主要活性成分亦为三七总皂苷,以人参皂苷Rb1、Rg1、Re和三七皂苷R1为主要成分。对于三七叶中有效成分的     

人参属植物高温蒸制前后人参皂苷含量的变化和细胞毒活性研究

目的研究人参属植物人参Panax ginseng、三七P. notoginseng和西洋参P. quinquefolium高温蒸制前后主要人参皂苷的含量变化,并测定高温蒸制前后样品对4株肿瘤癌细胞的细胞毒活性。方法采用HPLC建立了测定22种皂苷含量的分析方法,测定这些皂苷在人参属植物及其高温蒸制品中的含量。用MTT法测定人参属植物及其高温蒸制品对4株人类癌细胞(人类骨髓癌HL-60细胞、肝癌SMMC-7721细胞、肺癌A-549细胞、乳腺癌SK-BR-3细胞)的细胞毒活性。结果从人参、三七和西洋参及高温蒸制后的样品中鉴定出22个皂苷,包括人参皂苷Rg1、Re、Rb1、Rc、Rb2、Rd、Rk3、Rh4、Rk1、Rg5、Rb3、Rh3、Rk2,20(S)-人参皂苷Rh1、20(R)-人参皂苷Rh1、三七皂苷Fc、三七皂苷R1、绞股蓝皂苷IX、20(S/R)-三七皂苷Ft1、20(S/R)-人参皂苷Rg3、人参皂苷Rs3、人参皂苷Rh2。人参皂苷Rg1、Re、Rb1、Rc、Rb2和Rd为人参的主要成分;人参皂苷Rg1、Re、Rb1和Rd为西洋参的主要成分;人参皂苷Rg1、Re、Rb1、R...     

三七总皂苷对过氧化氢损伤人脐静脉内皮细胞的保护作用研究

目的:观察三七总皂苷(PNS)对过氧化氢损伤人脐静脉内皮细胞(HUVECs)LDH释放量、胞内SOD活性的影响,研究其对受损细胞粘附分子ICAM-1及MCP-1 mRNA表达的影响,探讨三七总皂苷对HUVECs的保护作用及其在动脉粥样硬化中作用的可能机制。方法:三七总皂苷预孵育细胞4h后,加入300μmol/L H2O2,继续培养24h,MTT法检测细胞活力,采用试剂盒测定LDH释放量、SOD酶活力,荧光实时定量PCR(RT-PCR)法检测三七总皂苷作用下内皮细胞ICAM-1及MCP-1 mRNA的表达。结果:三七总皂苷在25~100μg/ml浓度范围内可显著提高受损细胞活力,并能显著降低LDH漏出率,提高胞内SOD酶活力;RT-PCR结果表明H2O2组内皮细胞表达的ICAM-1和MCP-1 mRNA明显高于正常细胞组,而同时加入50及100μg/ml的三七总皂苷后两者mRNA表达明显下降。结论:三七总皂苷可保护血管内皮细胞免受H2O2的损害,改善受损细胞氧化指标,降低参与动脉粥样硬化相关细胞因子表达,从而预防和治疗动脉粥样硬化。     

广西三七皂苷类成分含量的分析

目的研究广西三七不同产地、不同部位的皂苷含量特征。方法利用香草醛-高氯酸比色法测定三七中总皂苷含量,以高效液相色谱（HPLC）法测定三七中人参皂苷Rg1,Rb1,Re和三七皂苷R1含量。结果广西靖西县不同产地三七样品的总皂苷及单体皂苷含量存在差异;不同部位间皂苷含量的差异达到极显著水平,以根茎的最高,主根次之;同一样品中,各单体皂苷含量以人参皂苷Rg1最高,人参皂苷Re最低,不同样品中各单体皂苷间比值存在差异。结论广西靖西县各地三七质量存在差异,药材中人参皂苷Rg1、人参皂苷Rb1和三七皂苷R1三者含量之和符合2005版《中国药典》规定。     

薄层扫描法测定三七药材中人参皂苷Rg1的含量

中药三七为五加科植物三七的干燥根。皂苷类是其主要生理活性成分,文献报道三七总皂苷含量约12 % [1～3] 。三七的薄层指纹图谱中,较明显的斑点为人参皂苷Rg1,Re ,Rb1及三七皂苷R1。人参皂苷Re与三七皂苷R1斑点极为接近,人参皂苷Rb1的Rf 值偏低,只有人参皂苷Rg1的斑点最清晰,分离效果好,故参考文献[4 ]采用双波长薄层扫描法测定三七药材中人参皂苷Rg1的含量。1　仪器、试剂和药材CS 930型薄层扫描仪(日本岛津公司) ,硅胶G薄层板....     

根腐对三七的皂苷含量影响

目的:对比分析根腐三七与正常三七中总皂苷及R1、Rg1、Rb1三种单体的含量,为根腐三七的质量评价及利用提供依据.方法:同田对比正常三七、根腐三七及不同腐烂程度三七的皂苷含量与单体成分,用HPLC法测定单体皂苷含量,大孔吸附树脂吸附比色法测定总皂苷含量.结果:三七根腐后比正常三七总皂苷含量平均降低了30.88%,R1、Rg1、Rb1三种单体平均含量分别降低了25.53%、11.26%、17.82%.结论:三七的皂苷含量随根部腐烂程度的增加而减少,根部腐烂对R1、Rg1、Rb1三种单体皂苷的比例无显著影响.     

HPLC法测定活血止痛片中三七皂苷R1、人参皂苷Rg1、Re、Rb1含量

目的：建立活血止痛片中三七皂苷R1、人参皂苷Rg1、Re、Rb1的含量测定方法。方法：采用HPLC法测定三七皂苷R1、人参皂苷Rg1、Re、Rb1的含量。结果：三七皂苷R1、人参皂苷Rg1、Re、Rb1四者的平均加样回收率分别为98.89%、97.73%、99.58%、99.18%,RSD分别为1.42%、1.02%、1.51%、1.32%。结论：该方法简便、灵敏,重现性好,可作为活血止痛片中三七皂苷R1、人参皂苷Rg1、Re、Rb1的含量测定方法。     

基于HPLC法和灰色关联法评价不同产地、不同商品规格三七质量

目的客观评价三七内在质量,构建三七质量评价的灰色关联模型,为三七质量评价及多指标成分中药材质量评价提供参考。方法采用HPLC法同时测定不同产地、不同商品规格三七样品中三七皂苷R1,人参皂苷Rg1、Re、Rb1、Rd含量,结合灰色关联法建立整体质量评价模型。结果所建立方法同时测定了三七皂苷R1,人参皂苷Rg1、Re、Rb1、Rd成分含量,方法学考察结果良好;灰色关联法可以区分出道地产区样品、合格样品和不合格样品。结论所建立的HPLC测定方法简单准确、稳定快速、分离效果好,可用于三七皂苷R1及人参皂苷Rg1、Re、Rb1、Rd的含量测定;灰色关联法可用于多指标成分中药质量评价。     

三七总皂苷治疗脑血管疾病研究进展

三七具有化瘀止血、消肿止痛等功效,三七总皂苷为其主要有效部位,主要成分包括人参皂苷Rb1、Rg1、Re、Rd及三七皂苷R1等。以三七总皂苷为药用物质基础的注射剂已成为治疗心脑血管疾病中药注射剂的主要品种,临床疗效确切。本文综述三七总皂苷治疗脑血管疾病的药理作用、临床应用及不良反应的研究进展,为深入研究及临床应用提供参考。     

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??OBJECTIVE To study the protective effects and mechanisms of aqueous extract from Descurainia sophia on doxorubicin(Dox)-induced cardiomyocyte injury.METHODS The Dox induced H9c2 cell apoptotic model was established, then the cells were divided into normal group (NC), model group (Dox), positive group (resveratrol,RSV), and different doses of aqueous extract of Descurainia sophia (DS). The viability of H9c2 cells was detected by MTT assay. The apoptosis rate, mitochondrial membrane potential and reactive oxygen species (ROS) level were detected by flow cytometry. The levels of T-SOD, LDH, MDA and GSH-PX were measured. And the protein expression levels of caspase-3, Bcl-2, Bax and p53 were detected by western blot, HPLC-MS identification of aqueous extract from Descurainia sophia.RESULTS After treating with DS products, the survival rate of H9c2 was increased (P<0.01), the apoptosis rate was significantly decreased (P<0.01),mitochondrial membrane potential was significantly increased (P<0.01), the level of ROS was significantly decreased (P<0.05), T-SOD and GSH-PX activities were significantly increased (P<0.01), the levels of LDH and MDA content were significantly decreased(P<0.01). Moreover, DS reduced the expression of caspase-3 (P<0.01), regulated the expression of Bax/Bcl-2 (P<0.01), decreased the expression of p53 (P<0.05). Seven components were identified from DS by HPLC/MS analysis.CONCLUSION DS can effectively protect cardiomyocyte, and its mechanism is probably associated with correcting functional disorders of oxidative stress of cardiomyocytes, and inhibit mitochondrial apoptotic pathway.     

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??OBJECTIVE To investigate the effect and the mechanism of baicalin in alleviating human umbilical vein endothelial cell (HUVEC) injury induced by lipocalin-2. METHODS Lipocalin-2 at different concentration (0, 5, 10, 20 ??mol??L^-1) and different time gradient (0, 24, 48 and 72 h) were added in HUVEC, the proliferation and apoptosis of HUVEC, contents of interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1) in HUVEC supernatant, lactic dehydrogenase (LDH) activities and the expressions of p-JNK, Bax and Bcl-2 in HUVEC were measured by MTT assay, flow cytometry, enzyme-linked immuno-sorbent assay (ELISA), colorimetry and Western blot, respectively. Different concentration of baicalin (15??30??60 mg??L^-1) or 10 ??mol??L^-1 SP600125, the specific inhibitor of JNK pathway, was added into HUVEC to detect the effect of baicalin. RESULTS Compared with the control group, 10 ??mol??L^-1 lipocalin-2 could significantly increase the apoptosis, IL-6, MCP-1 contents and LDH activities, p-JNK expression and Bax/Bcl-2 rate but restrain the proliferation in HUVEC (P<0.05). Compared with lipocalin-2 group, 30 mg??L^-1 baicalin could significantly restrain the apoptosis, IL-6, MCP-1 contents and LDH activities, p-JNK expression and Bax/Bcl-2 rate but increase the proliferation in HUVEC (P<0.05). CONCLUSION Our findings indicate that baicalin could alleviate HUVEC injury induced by lipocalin-2, the protective mechanism is related to the inhibition of JNK pathway.     

蛹虫草活性成分的提取分离及体外抗氧化作用研究

目的 以抑制蛋白酪氨酸磷酸酶1B(PTP1B)活性为导向,筛选蛹虫草子实体有效部位,分离纯化单体化合物,并检测其体外抗氧化作用。方法 通过硅胶色谱柱,半制备高效液相等色谱技术对蛹虫草子实体进行分离纯化,检测各组分对PTP1B的抑制活性;应用核磁碳谱、氢谱数据分析鉴定单体化合物结构;利用MTT法检测单体化合物对PC12细胞的增殖作用,及其对过氧化氢(H₂O₂)损伤的PC12细胞存活率的影响,试剂盒检测单体化合物对细胞乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量的影响。结果 发现5个对PTP1B具有较强抑制作用的活性成分,将其中活性最高的成分进一步分离纯化,获得单体化合物,经鉴定为β-D-吡喃葡萄糖基-9-甲基-4,8鞘氨醇(5),其对PTP1B具有较强的抑制活性,半抑制浓度(IC₅₀)为(3.42±0.59) μmol·L^-1。该单体化合物对H₂O₂诱导的PC12细胞氧化损伤具有明显的保护作用。结论 首次以抑制PTP1B活性为导向,在蛹虫草中分离得到脑苷脂类单体化合物,其具有抑制PTP1B作用和体外抗氧化活性,为蛹虫草用于糖尿病的预防和治疗奠定理论基础。     

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??OBJECTIVE To enhance the anticancer activity of doxorubicin(DOX) by conjugating DOX and vitamin E succinate (VES) and loading the conjugate into hyaluronic acid-octadecylamine (HA-C₁₈) copolymer micelles.METHODS DOX and VES were conjugated by amide reaction.DOX-VES/HA-C₁₈micelles were prepared via a probe-type ultrasonication technique.The morphology of the micelles was determined using a transmission electron microscopy (TEM).Dynamic light scattering (DLS) technique was used to determine the particle size distribution, hydrodynamic diameters, and stability of the micelles.Ultracentrifugation was exploited for measuring the drug loading (DL) and encapsulation efficiency (EE), and the in vitro release was investigated using a dialysis tubing.The cellular uptake and cellular distribution of drug-loaded micelles in MCF-7 cells were observed by fluorescence microscope, and the fluorescence intensity of DOX was evaluated by flow cytometer.The cytotoxicity of free DOX and drug-loaded micelles was tested by MTT assay against MCF-7 cells.RESULTS DOX-VES/HA-C₁₈ showed a nearly spherical morphology and good stability in PBS (pH 7.4) and 10% FBS.The particle size and zeta potential were (184.6??9.42) nm and (-20.7??1.23) mV, respectively.The DL and EE were (15.8??2.85)% and (94.2??1.32)%, respectively.DOX-VES/HA-C₁₈had a good controlled drug release property.Furthermore,DOX-VES/HA-C₁₈with accumulation in nucleipresented higher anti-tumor activity than free DOX and DOX/HA-C₁₈. CONCLUSION DOX-VES conjugate has synergistic anti-tumor effect and good application prospects.     

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??OBJECTIVE To investigate the protection of lyophilized powder of catalpol and puerarin (C-P) on oxygen-glucose deprivation/reperfusion(OGD/R)-injured astrocytes and the possible mechanism in vitro.METHODS Primary astrocytes were isolated from the cerebral cortex of neonatal rats. The purity of astrocytes was identified by GFAP immunofluorescence. Astrocytes were divided into 6 groups:normal group, model group, excipients group, and three groups with gradient doses of C-P (12.25, 24.50, 49.00 ??g??mL^-1). After astrocytes suffered from OGD/R (OGD 6 h/R 12 h) with or without C-P, cell survival, LDH leakage, and apoptosis were analyzed by MTT, colorimetry, and TUNEL respectively. The apoptotic protein, caspase-3, was evaluated by Western blot. Meanwhile, oxidative indexes including SOD, GSH, ROS, MDA, and NO were detected by assay kits. In terms of inflammation, TNF-??, IL-1??, and PGE₂ in medium were evaluated via ELISA. iNOS, COX-2, NF-??B p65, p-NF-??B p65, I??B-??, and p-I??B-?? were examined by Western blot. RESULTS The purity of primary astrocytes was more than 97%. Compared with normal group, the cell survival rate of model group decreased significantly, while the LDH leakage rate and cell apoptosis rate markedly increased after OGD/R injury in model group. Meanwhile, SOD activity and GSH level in astrocytes markedly decreased. The level of MDA and ROS significantly increased. The content of NO, TNF-??, IL-1??, and PGE₂ released in medium also sharply increased. However, those changes of related biochemical indexes above could be reversed by different gradient doses of C-P. There was no significant difference between excipients group and model group. Further study found that C-P could inhibit the protein expression of caspase-3, iNOS, and COX-2, as well as the increase of phosphorylation level of NF-??B p65 and I??B-?? significantly. CONCLUSION C-P shows a protective effect on the OGD/R injured astrocytes in vitro, and its protection mechanism involves in anti-inflammation, antioxidant, inhibiting the cell apoptosis and activation of NF-??B signaling pathway.     

锯叶棕提取物增强多烯紫杉醇抗MM细胞增殖作用

[目的]探讨锯叶棕提取物对多烯紫杉醇抗MM细胞增殖的影响。[方法]将体外培养的U266和PRMI8226细胞与0-2μL/mL的不同浓度锯叶棕提取物共孵育,应用MTT比色法测定细胞增殖活性;同时应用台盼蓝不相容实验检测锯叶棕提取物与多烯紫杉醇单独使用或合并使用时的细胞成活率。[结果]锯叶棕提取物对U266细胞与RP-M18226细胞的增殖具有明显的抑制作用,抑制生长50%(ED_(50S))的有效剂量分别为1.2和0.7μL/mL;U266细胞用锯叶棕提取物或多烯紫杉醇预处理,其细胞成活率分别为79.7%或90.1%,两者联合应用细胞成活率为51.6%(P<0.001);RPMI8226细胞用锯叶棕提取物或多烯紫杉醇预处理,其细胞成活率分别为73.7%或80.1%,两者联合应用细胞成活率为55.6%(P<0.001)。[结论]锯叶棕提取物抑制MM细胞增殖并增强多烯紫杉醇抗MM细胞增殖作用。     

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??OBJECTIVE To screen the best combination of extractum of Robinia-living trametes and chemotherapy and investigate the action mechanism of Robinia-living trametes against the apoptosis of human gastric cancer cell MGC803. METHODS MGC803 Cells were treated with different concentrations of Robinia-living trametes and chemotherapy drugs (5-Fu and paclitaxel) in vitro. The inhibitory rate of cells was measured by MTT assay. Morphological changes were observed with inverted microscope. The apoptosis rate of MGC803 cells which were treated with combination of Robinia-living trametes(0.2 mgmL^-1) and 5-Fu (2.5 ??gmL^-1) was detected by FCM. The protein expression of P53 and p-Akt in MGC803 cells which were treated with combination of Robinia-living trametes (0.2 mgmL^-1) and 5-Fu (2.5 ??gmL^-1) was detected by Western blot. RESULTS The viability of MGC803 cells was reduced by Robinia-living trametes and chemotherapy drugs (5-Fu and paclitaxel) in a concentration- and time-dependent manner(P<0.01). Under reverse microscopy, cell body shrinking, nuclear pyrosis, and nuclear fragmentation were observed. The higher concentration, the longer treatment time, the more cells died. Compared with monotherapy, the combination of Robinia-living trametes and chemotherapy could reduce the survival rate of MGC803 cells. The protein expressions of P53 in MGC803 cells treated with combination of drugs was up-regulated, while that of P-Akt was down-regulated. CONCLUSION The apoptosis of MGC803 cells in vitro may be induced by the inhibitory effect of the combination of Robinia-living trametes and 5-Fu on PI3K/Akt signaling pathway. Combination therapy of Robinia-living trametes and 5-Fu is potentially more effective in inhibition of tumor cells than monotherapy of Robinia-living trametes.     

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??OBJECITVE To study the inhibitive effect of plumbagin on Lewis lung cancer. METHODS Cell proliferation was determined by CCK8 assay. Apoptosis was determined by flow cytometry. The expression of Bcl-2 and VEGF protein was studied by Western blot assay. The model of C57BL/6 mice bearing Lewis lung cancer was established by subcutaneous seeding of Lewis lung cancer cells, and randomly divided into 5 groups (n=6). Tumor-bearing mice were injected with normal saline, plumbagin(low, medium, high dose) or cyclophosphamide (CTX) in each group. The tumor volume was measured. All mice were sacrificed on Day 22nd under aseptic condition for the tumor collection. The transplanted tumors were weighed for calculation of the tumor inhibition rate; Immunohistochemical method was applied to assessing the VEGF expression in tumor tissue. RESULTS CCK-8 assay showed that plumbagin had an obvious inhibition on Lewis lung carcinoma cells line in a dose-dependent manner(r=0.953, P<0.05). Plumbagin significantly increased cell apoptosis rate(P<0.05). The protein levels of Bcl-2 and VEGF were significantly reduced by plumbagin (0, 2.5, 5, 10 ??mol??L^-1) treatment(P<0.05). In plumbagin(low, medium, high dose) groups and CTX group, the tumour volume, tumour weight and the expression of VEGF were significantly less than those in the control group (P<0.05). CONCLUSION The plumbagin effectively inhibits Lewis lung carcinoma cells proliferation and tumor growth of Lewis lung carcinoma cells in mice. The mechanism involved is down-regulating the expression of Bcl-2 ,VEGF and inducing cell apoptosis.     

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??OBJECTIVE To study the effects of 5-Fu combined with resveratrol (Res) on the growth and apoptosis of A431 and TE-1 cell lines and the underlying mechanism.To study the therapeutic effects of 5-Fu/Res combination on mouse skin papilloma chemically induced by DMBA/TPA. METHODS The effects of 5-Fu/Res combination on the viability of cancer cells were evaluated by MTT assay, growth curves assay and LDH releasing assay. The inhibitory effect of combination of the two drugs was analyzed by the method of Chou and Talalay. Apoptoses of A431 and TE-1 were determined by inverted microscope and gel electrophoresis of DNA fragment analysis. Intracellular Ca²⁺ concentration was determined to study the underlying mechanisms of 5-Fu and Res on the apoptosis of cancer cells. The mouse skin papilloma model was established by DMBA and TPA,the expression of actived-caspase-3 in mouse skin was examined by immunohistochemistry. RESULTS Combination of 5-Fu and Res decreased the median inhibitory concentration (IC₅₀) to cancer cells remarkably. 5-Fu/Res combination showed a synergistic effect on apoptosis of A431 and TE-1 cells. Much more typical morphological changes of apoptosis and amount of fragmented DNA were observed in the cells treated with 5-Fu and resveratrol in combination than that in the cells treated with the agents alone. The increase of [Ca²⁺]_i induced by 5-Fu/Res combination might be involved in the apoptotic induction. There was a significant decrease of the number of tumors after treatment with 5-Fu and Res in the tumor-bearing mice model. Active caspase-3 in the cells of mice skin was generated by 5-Fu in combination resveratrol was more effective than either alone. CONCLUSION The 5-Fu/Res combination shows synergistic anti-tumor effects both in vitro and in vivo.