辽东楤木总皂苷降血糖作用研究

目的测定辽东楤木总皂苷降血糖活性.方法:辽东楤木总皂苷以120,60,30mg·kg-1剂量连续灌胃给正常小鼠9d;以100,50,25mg·kg1剂量连续给药四氧嘧啶诱导高血糖家兔4周;以120,60,30mg·kg-1剂量单次给药链脲霉素致高血糖小鼠,并分别于给药后测定血糖值.结果:辽东楤木总皂苷连续给药120,60mg·kg-1两剂量组使受试正常小鼠血糖明显降低;总皂苷连续灌胃给药,可使四氧嘧啶高血糖家兔血糖值明显降低,且随给药时间延长其降糖作用日趋明显,停药后其降糖作用可维持2～4h;单次灌胃给药,三剂量组于给药后2、4h使链脲霉素诱导的高血糖小鼠血糖明显降低,与给药前比较有非常显著或显著差异.结论:辽东楤木总皂苷具有较好的降血糖活性.     

重楼总皂苷对人胃癌MKN-45细胞凋亡及Fas/FasL信号通路的影响

目的:研究重楼总皂苷(Paridis Rhizoma total saponins,PRTS)诱导人胃癌MKN-45细胞凋亡的机制.方法:在前期研究基础上,将PRTS设定3个不同质量浓度(5,10,20 mg·L-1)处理人胃癌细胞12h,采用荧光染色法观察MKN-45细胞凋亡形态学变化;流式细胞术(Annexin V/PI双染色法)测定细胞凋亡率;ELISA测定MKN-45细胞caspase-3,caspase-8酶活性;Western blotting法检测Fas,FasL蛋白表达水平.结果:在荧光显微镜下,经PRTS处理的MKN-45细胞,可见典型的凋亡形态学特征;PRTS可明显增加细胞凋亡率,且PRTS 20 mg· L-1与对照组相比凋亡率有显著差异(P＜0.01);PRTS作用于MKN-45细胞12h,与对照组比较,其caspase-3,caspase-8活性明显增高(P＜0.01),Fas,FasL蛋白表达明显增高(P＜0.01).结论:PRTS能显著诱导人胃癌细胞MKN-45凋亡,其机制可能与增加caspase-3,caspase-8活性,上调Fas和FasL的表达有关.     

竹节参总皂苷对脑缺血损伤大鼠海马一氧化氮合酶的影响

目的:探讨竹节参总皂苷(tRPJS)对脑缺血损伤的保护作用机制,观察药物对局灶性脑缺血大鼠和慢性前脑缺血大鼠海马组织一氧化氮合酶(NOS)及诱导型一氧化氮合酶(iNOS)的影响。方法:采用线栓法建立局灶性脑缺血大鼠模型,双侧颈总动脉永久结扎方法制备慢性前脑缺血,研究竹节参总皂苷对这两种脑缺血损伤大鼠模型海马组织NOS,iNOS的影响。结果:与假手术组相比,局灶性脑缺血大鼠海马组织NOS,iNOS含量均明显增高(P<0.05),竹节参总皂苷能显著降低局灶性脑缺血大鼠NOS,iNOS的含量(P<0.05);慢性前脑缺血大鼠与假手术组相比,海马组织总NOS活力没有明显改变,iNOS含量增高(P<0.05),竹节参总皂苷能显著降低慢性前脑缺血大鼠海马组织iNOS的含量(P<0.05)。结论:竹节参总皂苷抗缺血性海马神经元损伤与降低海马组织iNOS的活性有关。     

地榆总皂苷对巨核祖细胞增殖分化及相关受体表达的影响

目的:观察地榆总皂苷(DYS)对Baf3细胞和32D细胞的促增殖和分化作用,以及对IL-3受体和干细胞因子受体c-kit表达水平的影响。方法:培养依赖IL-3生长的Baf3细胞和32D细胞,在加或不加IL-3条件下,用质量浓度为5,10,20,30,40 mg·L-1的DYS分别处理细胞24,48,72,96 h后,采用CCK8方法检测细胞增殖;并用Giemsa染色检测细胞分化;另以RT-PCR方法检测DYS对Baf3细胞IL-3受体及对32D细胞c-kit表达水平的影响。结果:DYS单独处理细胞48 h,明显促进Baf3细胞和32D细胞增殖;与对照细胞比较,DYS处理32D细胞呈现成簇生长并形成许多大的细胞团;在加入IL-3的条件下,DYS也能明显增强IL-3的促细胞增殖作用。此外,DYS高浓度单独处理32D细胞可明显诱导多倍体巨核细胞数增加,促进巨核细胞分化。在mRNA水平,DYS单独处理细胞也能明显上调IL-3受体和c-kit的表达。结论:地榆总皂苷单独作用能促使2种巨核祖细胞的增殖和分化,其作用与上调IL-3受体和c-kit表达水平有关。     

竹节三七总皂苷对胶原诱导型关节炎大鼠抗氧化能力、细胞因子水平的影响

目的:研究竹节三七总皂苷(total saponins of Panax japonicus,TSPJ)对胶原诱导型关节炎大鼠抗炎、抗氧化功能的影响。方法:注射Ⅱ型胶原蛋白与完全费氏佐剂的乳化剂建立大鼠关节炎模型。观察致炎前后大鼠足容积的改变检测竹节三七总皂苷对关节炎模型大鼠的作用效果。检测关节炎大鼠抗氧化指标及血清细胞因子水平,观察竹节三七总皂苷对模型大鼠抗炎作用机理。结果:与模型组比较,25 mg/kg、50 mg/kg、100 mg/kg竹节三七总皂苷能减轻关节炎大鼠足爪肿胀度,100mg/kg组尤为明显;100mg/kg竹节三七总皂苷增加模型大鼠血中谷胱甘肽过氧化物酶、超氧化物歧化酶活性,抑制丙二醛水平;100mg/kg竹节三七总皂苷能降低模型大鼠血中白细胞介素-1、白细胞介素-8、肿瘤坏死因子-α水平,50mg/kg竹节三七总皂苷低模型大鼠血中白细胞介素-8水平效果明显。结论:竹节三七总皂苷具有抗炎作用,能增强关节炎大鼠的抗氧化能力,降低关节炎大鼠白细胞介素-1、白细胞介素-8、肿瘤坏死因子-α水平,可能是其治疗类风湿性关节炎的重要机制之一。     

三七总皂苷对模拟航渡及大强度运动大鼠神经-内分泌-免疫网络的影响

目的:观察模拟航渡及大强度运动对雄性大鼠神经-内分泌-免疫网络的影响及三七总皂苷的调控作用。方法:将6周龄的SD雄性大鼠随机分为静息对照组(QC组)、训练对照组(TC组)和三七总皂苷组(PNS组),每组10只。QC组不接受任何刺激,在生理盐水灌胃1周后即进行取样。TC组和PNS组大鼠经过10 d递增负荷跑台训练,分别生理盐水和PNS灌胃1周后,进行模拟航渡及高强度运动,结束即刻取血,使用放免法测定大鼠血清皮质酮(Cort)、促肾上腺皮质激素(ACTH)、β-内啡肽(β-EP)、神经肽Y(NPY)、白介素1β(IL-1β)和白介素6(IL-6);采用免疫组织化学方法检测各组大鼠垂体ACTH,NPY的表达水平,并进行相关分析;电镜下观察垂体、肾上腺超微结构。结果:训练对照组大鼠与静息对照组相比,血清Cort,β-EP水平显著升高,同时,垂体ACTH,NPY表达水平显著降低。灌服PNS后与TC组相比,血清Cort,ACTH及NPY水平下降;同时,免疫组化结果表明垂体ACTH,NPY表达水平也明显增高。在此基础上观察到应激后大鼠腺垂体及肾上腺皮质的超微结构改变,功能受损,灌服PNS后对细胞具有保护作用。结论:负性心理应激及大强度的运动训练使大鼠神经-内分泌-免疫网络功能紊乱,PNS对神经-内分泌-免疫网络具有调控作用。     

三七总皂苷对小鼠肝脏谷胱甘肽-S-转移酶的抑制作用及其动力学分析

目的:研究三七总皂苷(total saponins of Panax notoginseng,PNS)对小鼠肝脏谷胱甘肽-S-转移酶( glutathione-Stransferase,GST)的抑制作用及其酶动力学.方法:采用差速离心法制备小鼠胞质液酶,用半数效量概率单位法及LineweaverBurk双倒数作图法求得小鼠肝脏GST的米氏常数(Km)、最大反应速度(Vmax)和三七总皂苷对小鼠肝脏GST的半数抑制浓度( IC50)以及判断三七总皂苷对GST的抑制类型和抑制常数(KI,KIS).结果:三七总皂苷对小鼠肝脏GST有抑制作用,其IC50是189.54 mg·L-1;对底物还原型谷胱甘肽(GSH),GST的Km是0.456 3 mmol·L-1,Vmax是476.19 U·mg-1;对底物1-氯-2,4-二硝基苯(CDNB),GST的Km是0.109 7 mmol·L-1,Vmax是400.00 U·mg-1.对底物GSH,三七总皂苷对小鼠肝脏GST属于混合性抑制,其抑制常数KI是0.27 mg·L-1,KIS是0.68 mg·L-1:对底物CDNB,三七总皂苷对小鼠肝脏GST属于混合性抑制,其抑制常数KI是0.21 mg·L-1,KIS是0.66 mg· L-1.结论:三七总皂苷对小鼠肝脏GST的活性有明显抑制作用.     

金铁锁总皂苷抗炎镇痛作用及作用机理研究

目的：通过实验观察和探讨金铁锁总皂苷（TSPT）抗炎作用及其作用机理.方法：建立佐剂性关节炎（AA）动物模型,观察TSPT对AA大鼠炎性痛阈及炎性组织液中丙二醛（MDA）和皮质醇（Cor）水平的影响来探讨其镇痛抗炎作用及其作用机理.结果：TSPT能有效提高AA大鼠致炎足的痛阈,降低炎性组织液中MDA,双向调节AA大鼠血中Cor的水平.结论：TSPT具有良好的抗炎镇痛作用,其作用机理与降低MDA,调节Cor水平有关.     

RP-HPLC测定西洋参叶皂苷碱降解物中20(S)-人参皂苷Rh2的含量

目的:对西洋参叶皂苷碱降解物中20(S)-人参皂苷Rh2进行含量测定。方法:采用RP-HPLC,ZORBAXEXTEND C₁₈柱(4.6 mm×250 mm,5μm),流动相甲醇-水(85∶15),流速1.2 mL.min^-1,检测波长203 nm,柱温25℃。结果:20(S)-人参皂苷Rh₂的进样量在0.5～25μg有良好的线性关系,r=0.999 9;平均加样回收率为99.7%,RSD1.0%。结论:该方法测定20(S)-人参皂苷Rh2简便快捷,准确度高,测定结果证实该降解方法20(S)-人参皂苷Rh2的转化率高。     

茅莓总皂苷对局灶性脑缺血再灌注大鼠脑水肿及血-脑脊液屏障变化的影响

目的：研究茅莓总皂苷(TSRP)对局灶性脑缺血再灌注大鼠脑水肿及血脑屏障通透性的影响。方法：采用线栓法制备大鼠局灶性脑缺血再灌注模型,在大鼠脑缺血2 h再灌注6,24,48,72 h分别测定脑含水量和伊文思蓝含量的变化及茅莓总皂苷各剂量组的影响。结果：与模型组比较茅莓总皂苷各剂量组均能不同程度降低脑含水量和伊文思蓝含量。结论：茅莓总皂苷各剂量组显著降低缺血2h再灌注6,24,48,72 h脑水肿和其对血脑屏障通透性,这可能是其减轻I/R时脑水肿的机制之一。     

甘草皂苷与桔梗皂苷合用的时间协同研究

目的：考查甘草皂苷与桔梗皂苷祛痰和抗炎作用的时间协同。方法：采用气管酚红排泌、二甲苯致耳肿胀及PLA2致足跖肿胀模型,考查两者合用对药效持续时间的影响,以及运用金氏Q值法对两者合用的相互作用性质进行定量评价。结果：甘草皂苷与桔梗皂苷合用可明显延长药效持续时间,Q值法评价显示出不同结果,随着时间延长,在气管酚红排泌模型中,由相加转为协同;在二甲苯致耳肿胀模型中,单用无效合用有效的时间点下,表现为相加和协同;在PLA2致足跖肿胀模型中,则表现为相加和拮抗。结论：甘草皂苷与桔梗皂苷合用在祛痰与抗炎方面具备时间协同特点,协同性质与两药配比、剂量有关。     

基于计算机辅助设计的竹节参总皂苷改善HepG2细胞脂质代谢的作用及机制研究

目的研究竹节参总皂苷(TSPJ)改善人肝癌HepG2细胞脂质代谢的作用,基于计算机辅助药物设计,预测并验证TSPJ可能的作用靶点。方法应用棕榈酸(PA)诱导HepG2细胞脂肪变性模型,实验分为对照组、模型组、TSPJ(50.0、25.0、12.5 mg/L)组。5组细胞均培养24 h后,分别采用油红O染色和尼罗红染色定性、定量检测细胞内脂质积累情况;采用试剂盒检测细胞内三酰甘油(TG)的含量;采用计算机辅助药物设计预测TSPJ可能的作用靶点,用免疫荧光检测相关蛋白的表达。结果 PA浓度为100μmol/L处理细胞24 h可以造成HepG2细胞脂质积累模型,TSPJ可以显著改善HepG2细胞脂质积累(P0.01),降低TG含量;利用计算机辅助药物设计预测TSPJ的作用靶点可能是雌激素相关受体;与对照组比较,模型组雌激素β受体(ERβ)表达水平降低,与模型组比较,用药组ERβ表达水平均升高。结论 TSPJ可以显著改善HepG2细胞脂质代谢,其作用靶点可能是ERβ。     

The protective effects of total saponins from Ornithogalum saundersiae (Liliaceae) on acute hepatic failure induced by lipopolysaccharide and d-galactosamine in mice

Ethnopharmacological relevance

This study examined the protective effects of total saponins from Ornithogalum saundersiae (Liliaceae) on d-galactosamine (d-GalN) and lipopolysaccharide (LPS) - induced fulminant hepatic failure.

Materials and methods

Total saponins of Ornithogalum saundersiae (Liliaceae) (OC) were prepared with ethyl alcohol extract from bulbs of the plant. Mice were given an intraperitoneal injection of d-GalN (700 mg/kg)/LPS (10 μg/kg). OC (100 mg/kg, 200 mg/kg and 300 mg/kg) was administered orally for 3 days continuously, and at the last day at 1 h before the d-GalN/LPS injection. Mice were sacrificed at 8 h after the d-GalN/LPS injection. The liver injury was assessed biochemically, investigating aspartate aminotransferase (AST), alanine aminotransferase (ALT), malondialdehyde (MDA), glutathione (GSH) activities, and the expressions of caspase-3 and hypoxia inducible factor-1α (HIF-1α) as well. Tumor necrosis factor (TNF-α) content was measured after d-GalN/LPS induced 1 h by ELISA assay. The survival rates after application of OC in 24 h also were observed.

Results

d-GalN/LPS increased the serum aminotransferase levels and lipid peroxidation, while decreased the reduced glutathione level. The pretreatment with OC attenuated these changes in a dose-dependent manner. Elevation of TNF-α level and activation of caspase-3, HIF-1α were observed in the d-GalN/LPS group, which was attenuated by OC. The survival rate of the OC groups was significantly higher than that of the d-GalN/LPS group.

Conclusions

Protection afforded by OC against d-GalN/LPS-induced fulminant hepatic failure is the result of reduced oxidative stress, inhibited expression of caspase-3, HIF-1α, and anti-apoptotic activity.     

Effect of total saponins of Panax notoginseng (TSPNS) on myocardial intracellular Ca2+ and activity of calcium pump of membrane of sarcoplasmic reticulum in SHR]

A comparative study was conducted on the action of TSPNS, NS and Captopril in treating SHR, with WKY of similar ages taken as normal control. The results demonstrate that TSPNS helps increase the effect of calcium pump on the membrane of sarcoplasmic reticulum, decrease the myocardial intracellular Ca2+ and reduce the mass of the left ventricular muscle.     

Effect of Bridelia ferruginea Benth (Euphorbiaceae) ethyl acetate and acetone fractions on insulin resistance in fructose drinking mice

Ethnopharmacological relevance

The root of Bridelia ferruginea is traditionally used as a treatment for type 2 diabetes. The present study was investigated to evaluate the effect of Bridelia ferruginea root bark fractions on some markers of type 2 diabetes on fructose drinking mice.

Materials and methods

Mice received a solution of fructose 15% during 42 days ad libitum; at the 15th day to the 42nd day, they received distilled water for fructose drinking control group, metformin 50 mg/kg per day or fractions 50 mg/kg per day for treatment groups. The normal control group received only distilled water during the experiment. After 6 weeks of experiment, OGTT, fasting blood glucose, plasma insulin, triglycerides (TG), total cholesterol, AST and ALT levels were measured.

Results

Fructose drinking control group (F) showed significant (p<0.001) increase of glucose tolerance, plasma levels of total cholesterol, triglycerides and insulin index for insulin resistance (Homeostasis Model Assessment ratio HOMA-IR) as compared to normal control mice. In treated groups, there was a significant reduction of glucose intolerance respectively 74% (p<0.001), 25% (p<0.5) and 92% (p<0.001) for ethyl acetate fraction, acetone fraction and metformin at the same dose of 50 mg/kg per day during 4 weeks administration. In ethyl acetate fraction and metformin treated groups, biochemical parameters and insulin index were significantly (p<0.001) lower than that of fructose drinking control group.

Conclusions

This indicates that Bridelia ferruginea root bark ethyl acetate fraction improved insulin resistance as metformin significantly in type 2 diabetes.     

Total saponins of Panax ginseng (TSPG) promote erythroid differentiation of human CD34 cells via EpoR-mediated JAK2/STAT5 signaling pathway

Ethnopharmacological relevance

Total saponins of Panax ginseng (TSPG), main constituents extracted from Panax ginseng, a highly valued traditional Chinese medicine, have been shown to be an effective agent on hematopoiesis.

Objective

To investigate the effect and mechanism underlying in which TSPG promote human CD34⁺ hematopoietic stem and progenitor cells to differentiate into erythroid-lineage cells.

Materials and methods

The effect of TSPG on erythroid differentiation of purified CD34⁺ cells derived from umbilical cord blood (UCB) was determined by methylcellulose assay system and colorimetry for hemoglobin content. The changes of EpoR expression in umbilical cord blood mononuclear cells (UCB-MNCs) and purified CD34⁺ cells were detected with Western blotting and flow cytometry, respectively, and observed under laser scanning confocal microscope (LSCM). RT-PCR was performed to examine EpoR mRNA expression in CD34⁺ cells. The effects of TSPG-pretreatment on Epo-induced JAK₂ and STAT₅ tyrosine phosphorylation were analyzed by immunoprecipitation.

Results

The addition of TSPG (20–70 mg/L) increased the colony formation rate of BFU-E. TSPG (50 mg/L) alone used significantly increased the hemoglobin content, the addition of AG490 evidently reduced TSPG-induced elevation of hemoglobin content. TSPG increased the expression of EpoR on the surface membrane of CD34⁺ cells but did not change the expression of EpoR in total UCB-MNCs. TSPG also increased the expression of EpoR mRNA in CD34⁺ cells. TSPG markedly enhanced Epo-induced tyrosine phosphorylation of JAK₂ and STAT₅ in UCB-MNCs.

Conclusion

These findings suggest that TSPG may enhance the erythroid differentiation of hematopoietic stem and progenitor cells via Epo/EpoR-mediated JAK₂/STAT₅ signaling pathway.     

2株内生真菌对菊花抗旱特性的影响

目的:以PEG6000模拟干旱条件,研究接种内生真菌(葡萄孢属C1菌Botrytis sp.、球毛壳菌G4菌Chaetomium globosum对药用菊花Ch.morifolium抗旱性的影响.方法:分别用0%,10%,20%,30%,40%PEG6000胁迫菊花组培苗4 d,测定各处理组菊花生物量,叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)活性及叶片丙二醛(MDA)、可溶性蛋白含量.结果:模拟干旱胁迫后,接种内生真菌的菊花长势好于对照(未接菌),PEG6000胁迫4 d后,随着胁迫浓度的增加,菊花总生物量不断减少,接菌组生物量显著高于对照,C4组高于C1组;各处理组MDA含量随着胁迫浓度的增加而不断增加;各处理组POD活性、可溶性蛋白含量随着胁迫浓度的增加均呈现先增加后减小的趋势;对照SOD活性随着胁迫浓度的增加而逐渐增加,接菌组SOD活性基本保持不变;对照PAL活性随着胁迫浓度的增加而逐渐增加.不同浓度PEG胁迫后,接菌组MDA含量始终低于对照,SOD活性、POD活性、可溶性蛋白含量、PAL活性均始终高于对照.结论:接种内生真菌提高菊花的抗旱能力.     

去雄黄牛黄解毒片抗菌及解热作用研究

去雄黄牛黄解毒片体外抑菌试验对金葡菌和甲型链球菌最敏感,其MIC分别为7.8和15.6mg/ml;体内抗菌试验ig1.7和0.8g/kg,对乙型链球菌感染的小鼠有明显的保护作用,可使死亡率明显减少。本品尚有明显的解热作用,能明显对抗霍乱菌苗和有基酚引起的家兔和大鼠体温升高,上述作用与等剂量的牛黄解毒片相当。     

基于药物体系质量评价模式的柴胡质量表征关联分析研究

目的：运用药物体系质量评价模式对柴胡饮片进行质量表征及关联分析,以有效地评价柴胡饮片的质量。方法：采用HPLC法同时测定柴胡饮片中药物体系关联成分芦丁、柴胡皂苷c、柴胡皂苷a、柴胡皂苷d的含量,同时表征主要黄酮类成分含量和及主要皂苷类成分含量和,并采用可见分光光度法测定柴胡饮片中总酚类成分含量。结果：基于药物体系质量评价模式,对柴胡饮片中有效指标性成分含量及其相对比值进行质量表征,同时将不同批次的饮片与经过药效验证的基准饮片进行关联度分析。结果表明,样品7、8、14中有效指标性成分含量总体高于基准批号5,批号6中有效指标性成分含量与基准批次饮片5相近,样品7、8、13、14与基准批次饮片5关联度较高。综合质量表征及关联分析结果,样品7、8、14、5、13的品质居优。结论：基于药物体系质量评价模式对柴胡饮片的质量进行评价,通过有效指标性成分含量及其相对比值的质量表征,以经过药效验证的饮片为基准,结合关联度分析,可综合全面地评价柴胡饮片的质量,为柴胡饮片的资源筛选、药物原料的质量控制和应用提供依据,同时为中药质量评价提供了方法学及其应用借鉴。