Effects of total saponins of Panax notoginseng on immature neuroblasts in the adult olfactory bulb following global cerebral ischemia/reperfusion

The main active components extracted from Panax notoginseng are total saponins. They have been shown to inhibit platelet aggregation, increase cerebral blood flow, improve neurological behavior, decrease infarct volume and promote proliferation and differentiation of neural stem cells in the hippocampus and lateral ventricles. However, there is a lack of studies on whether total saponins of Panax notoginsertg have potential benefits on immature neuroblasts in the olfactory bulb following ischemia and reperfusion. This study established a rat model of global cerebral ischemia and reperfusion using four-vessel occlusion. Rats were administered total sa- ponins of Panax notoginseng at 75 mg/kg intraperitoneally 30 minutes after ischemia then once a day, for either 7 or 14 days. Total saponins of Panax notoginseng enhanced the number of dou- blecortin （DCX）＋ neural progenitor ceils and increased co-localization of DCX with neuronal nuclei and phosphorylated cAMP response element-binding/DCX＋ neural progenitor cells in the olfactory bulb at 7 and 14 days post ischemia. These findings indicate that following global brain ischemia/reperfusion, total saponins of Panax notoginseng promote differentiation of DCX＋ cells expressing immature neuroblasts in the olfactory bulb and the underlying mechanism is related to the activation of the signaling pathway of cyclic adenosine monophosphate response element binding protein.

Total saponins of Panax ginseng effects on proliferation and differentiation of human embryonic neural stem cells and in a Parkinson's disease mouse model

BACKGROUND： Total saponins of Panax ginseng （TSPG） exhibits neuroprotection against Parkinson＇s disease in the substantia nigra. OBJECTIVE： To investigate the effects of TSPG on human embryonic neural stem cells （NSCs） proliferation and differentiation into dopaminergic neurons using in vitro studies, and to observe NSC differentiation in a mouse model of Parkinson＇s disease, as well as behavioral changes before and after transplantation. DESIGN, TIME AND SETTING： In vitro neural cell biology trial and in vivo randomized, controlled animal trial were performed at the Institute of Basic Medical Sciences, Chongqing Medical University between September 2004 and December 2007. MATERIALS： TSPG （purity 〉 95%） was isolated, extracted, and identified by Chongqing Academy of Chinese Materia Medica. Recombinant human basic fibroblast growth factor （bFGF） and recombinant human epidermal growth factor （EGF） were purchased from PeproTech, USA. A total of 25 C57/BL6J mice, aged 18-20 weeks were included. Twenty were used to establish a Parkinson＇s disease model with i.p. injection of MPTP （1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine） and TSPG alone or combined with interleukin-1 （IL-1）-treated NSCs prior to transplantation into the corpus striatum. The remaining five mice were pretreated for 3 days with TSPG prior to MPTP injection, serving as the TSPG prevention group. METHODS： Primary NSCs were isolated, cultured and purified from embryonic cerebral cortex. Immunocytochemistry was employed to detect specific antigen expression in the NSCs. In vitro experiment： （1） to induce proliferation, NSCs were treated with TSPG, EGF＋bFGF, or TSPG＋EGF＋bFGF, respectively; （2） to induce dopaminergic neuronal differentiation, NSCs were treated with TSPG, IL-1, or TSPG＋IL-1, respectively. MAIN OUTCOME MEASURES： In vitro experiment： the effects of TSPG on NSCs proliferation were evaluated with flow cytometry and MTT assay. Tyrosine hydroxylase expression was determined by immunocytochemistry assay to observe effects of TSPG on dopaminergic neuronal differentiation. In vivo experiment： differentiation of grafted NSCs in the mouse brain was determined by immunohistochemical staining. Behavioral changes were evaluated by spontaneous activity frequency, memory function, and score of paralysis agitans. RESULTS： （1） NSCs were cultured and passaged for more than three passages. Immunocytochemistry revealed positive nestin staining, as well as neurofilament protein and glial fibrillary acidic protein. （2） TSPG significantly increased NSC proliferation, in particular when combined with EGF and bFGF, which was twice as effective as FGF or bFGF alone. TSPG also induced dopaminergic differentiation in NSCs, in particular when TSPG was added together with IL-1, resulting in an effect five times greater than that of IL-1 alone. （3） At day 30 following transplantation, most NSCs in the TSPG prevention group differentiated into dopaminergic neurons, and the scores of paralysis agitans, spontaneous activity, and memory function were significantly increased compared with TSPG alone or TSPG＋IL-1 groups （P 〈 0.05）. CONCLUSION： TSPG stimulated NSC proliferation, in particular when combined with FGF and bFGF. TSPG significantly induced dopaminergic neuronal differentiation of NSCs, and the effect was greater when combined with IL-1. In addition, TSPG greatly improved behavior in the Parkinson＇s disease mouse model following NSC transplantation. Following NSC transplantation, TSPG pretreatment exhibited superior efficacy over either TSPG alone or TSPG in combination with IL-1, in terms of behavioral improvements in the Parkinson＇s disease mouse model.

Advances in Researches of Extraction, Separation, and Purification Technologies for Total Saponins of Panax notoginseng

Total saponins of Panax notoginseng have the functions of promoting blood circulation and removing phlegm, thus they have high medicinal value. There are many different extraction methods in the extraction and separation of total saponins of P. notoginseng . The extraction methods of total saponins of P. notoginseng are mainly divided into traditional extraction methods, modern extraction methods and compound extraction methods.

Advances in Researches of Extraction Processes of Total Saponins of Pnanx Notoginseng

Traditional extraction methods of total saponins of Panax notoginseng include cold soaking method,water decoction method,alcohol reflux method,percolation method,macroporous resin adsorption method,and accelerated solvent extraction( ASE) method. Modern extraction methods include ultrasonic extraction,microwave assisted extraction,supercritical CO_2 extraction,microbial fermentation assisted extraction,neural network model optimized extraction method,and multi-stage countercurrent extraction method. This paper discussed principles of these methods and compared their advantages and disadvantages.

三七总皂苷复合纳米囊泡对糖尿病足溃疡大鼠皮肤创面愈合及血管生成的影响

目的:探究三七总皂苷(PNS)复合纳米囊泡对糖尿病足溃疡(DFU)大鼠皮肤创面愈合及血管生成的影响。方法:取雄性SD大鼠构建DFU模型,将造模成功的27只大鼠随机分为DFU组、PNS组和PNS复合纳米囊泡(PNS-CNV)组,每组9只。另取12只血糖正常的皮肤创面大鼠作为对照组(Control组)。比较各组创面愈合率。采用酶联免疫吸附试验(ELISA)法检测大鼠血清中炎症因子白介素(IL)-6、IL-8和肿瘤坏死因子-α(TNF-α)水平,苏木精—伊红(HE)染色法检测创面组织病理学变化,免疫组化染色检测创面组织微血管密度(MVD),western blotting法检测创面组织中血管生成因子(VEGF)、神经源性基因同源蛋白(Notch1)蛋白表达。结果:造模后DFU组、PNS和PNS-CNV组大鼠空腹血糖(FBG)均高于Control组(P<0.05);药物干预7 d、14 d时,与DFU组相比,PNS组和PNS-CNV组大鼠FBG明显降低,且PNS-CNV组低于PNS组(均P<0.05)。与Control组相比,DFU组大鼠创面愈合率、创面组织CD34阳性表达、MVD及VEGF蛋白表达量均明显降低,血清IL-6、IL-8、TNF-α含量及创面组织Notch1蛋白表达量升高(均P<0.05);与DFU组相比,PNS和PNS-CNV组大鼠创面愈合率、创面组织CD34阳性表达、MVD及VEGF蛋白表达量升高,血清IL-6、IL-8、TNF-α含量及创面组织中Notch1蛋白表达量降低(均P<0.05);与PNS组相比,PNS-CNV组大鼠创面愈合率、创面组织CD34阳性表达、MVD及VEGF蛋白表达量升高,血清IL-6、IL-8、TNF-α含量及创面组织Notch1蛋白表达量降低(均P<0.05)。结论:PNS-CNV可上调DFU大鼠创面组织中VEGF表达,抑制炎症反应,促进新生血管生成,从而加速创面的愈合,且该作用优于PNS。

三七总皂苷对脑缺血再灌注后神经功能恢复的影响

目的探究三七总皂苷对脑缺血再灌注后神经功能恢复的影响。方法培养30只SPF级雄性SD大鼠,构建脑缺血再灌注模型。将大鼠分为假手术组、模型组和三七总皂苷组,每组10只大鼠。检测各组大鼠脑梗面积、脑含水量。Longa评分评价大鼠神经功能缺损情况。水迷宫实验检测大鼠神经功能。检测各组大鼠血清中氧化应激指标超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)、谷光甘肽(glutathione,GSH)含量和炎症因子指标肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-10(interleukin-10,IL-10)含量。Tunel染色检测各组大鼠海马神经元细胞凋亡情况。结果和假手术组大鼠相比,模型组大鼠脑梗面积、脑含水量、神经功能缺损评分、学习潜伏期和记忆潜伏期显著升高,SOD、GSH、IL-10含量显著降低,MDA、TNF-α、IL-1β含量显著升高,海马神经元凋亡比例显著升高(P<0.05)。和模型组大鼠相比,三七总皂苷组大鼠脑梗面积、脑含水量、神经功能缺损评分、学习潜伏期和记忆潜伏期显著降低,SOD、GSH、IL-10含量显著升高,MDA、TNF-α、IL-1β含量显著降低,海马神经元凋亡比例显著降低(P<0.05)。结论三七总皂苷可能通过抑制大鼠炎症反应和氧化应激,减少海马神经元凋亡,促进脑缺血再灌注后神经功能恢复。

三七总皂苷超声波经皮药物导入联合综合疗法治疗冈上肌肌腱炎的临床效果

目的:观察三七总皂苷超声波经皮药物导入联合综合疗法治疗冈上肌肌腱炎的临床效果。方法:选取2022年1月-2023年9月北京积水潭医院贵州医院收治的冈上肌肌腱炎患者60例作为研究对象,采用随机数字表法分为试验组和对照组,各30例。两组均给予综合疗法,试验组接受三七总皂苷葡萄糖溶液超声波经皮药物导入治疗,对照组接受体外冲击波治疗。比较两组疼痛情况、肩关节功能。结果:治疗前,两组疼痛视觉模拟评分法评分比较,差异无统计学意义(P>0.05);治疗后,两组疼痛视觉模拟评分法评分低于治疗前,且试验组低于对照组,差异有统计学意义(P<0.001)。治疗前,两组Constant-Murley肩关节功能评分比较,差异无统计学意义(P>0.05);治疗后,两组Constant-Murley肩关节功能评分高于治疗前,且试验组高于对照组,差异有统计学意义(P<0.05)。结论:三七总皂苷超声波经皮药物导入联合综合疗法治疗冈上肌肌腱炎的临床效果较好,能够减轻疼痛,改善患者肩关节功能。

竹节参总皂苷缓解CCl_(4)诱导的大鼠急性肝损伤:基于调控PI3K/Akt/NF-κB信号通路

目的探讨土家族药用植物竹节参提取物总皂苷对CCl_(4)致急性肝损伤的保护作用及潜在的药理学机制。方法将6周龄SPF级雄性SD大鼠随机分为正常组、模型组、联苯双酯组(100 mg/kg)、竹节参总皂苷低、中、高(50、100、200 mg/kg)剂量组,各组8只,除空白组外,其余各组采用CCl_(4)诱导大鼠急性肝损伤模型,处理组于造模中给予药物灌胃干预。比较各组大鼠的血清谷草转氨酶(AST)、谷丙转氨酶(ALT)、总胆红素(TBil)和碱性磷酸酶(ALP)水平;HE染色观察肝组织病理学改变;免疫组化检测肝脏组织PI3K/Akt/NF-κB信号通路相关分子的表达;酶联免疫法测定肝脏组织总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)水平;蛋白免疫印迹法检测肝脏组织PI3K-Akt和SIRT6-NF-κB通路相关蛋白表达情况。结果网络药理学分析显示,竹节参总皂苷对急性肝损伤的有治疗作用,其关键的通路为PI3K/Akt等信号通路。血清学和酶联免疫学实验结果显示,与正常组相比,模型组大鼠血清和肝组织的AST、ALT、ALP、TBil和MDA明显增高(P<0.01),T-SOD和GSHPx水平显著降低(P<0.01);与模型组相比,各治疗组ALT、AST、ALP、TBIL和MDA水平显著降低(P<0.01),T-SOD和GSH-Px水平显著升高(P<0.01)。免疫组化结果显示,正常组大鼠肝组织细胞内未见p-NF-κB表达阳性,与正常组相比,模型组中p-NF-κB阳性表达则明显增加,阳性细胞数增多(P<0.01);与模型组比较,各治疗组阳性表达明显减少(P<0.01)。免疫印迹结果显示,相对于正常组,模型组PI3K、p-Akt蛋白的表达水平下降,p-NF-κB、TNF-α和IL-6蛋白的表达水平升高(P<0.05);与模型组相比,各治疗组PI3K、p-Akt和SIRT6蛋白的表达水平显著升高,p-NF-κB p65、TNF-α和IL-6的蛋白的表达水平显著降低(P<0.05)。结论竹节参总皂苷可以通过调节PI3K/Akt和NF-κB通路,有效减缓CCl_(4)诱导的大鼠急性肝损伤,发挥其抗炎、抗氧化应激和保护肝损伤的作用。

Effects of Organic and Medium and Trace Element Fertilizers on Yield and Quality of Panax notoginseng

[Objectives]This study aimed to investigate the effects of organic and medium and trace element fertilizers(Ca,Zn,B)on yield and quality of Panax notoginseng to provide theoretical support for rational fertilization in cultivation of P.notoginseng.[Methods]Five fertilization treatments,control(CK),organic fertilizer(OM),zinc fertilizer(ZF),boron fertilizer(BF)and lime(LF),were designed.A two-consecutive-year field plot trail was conducted.The biological traits,yield and saponin content of P.notoginseng were determined.[Results]The application of organic fertilizer had no significant effect on the biological traits of P.notoginseng.Trace element fertilizers significantly increased the scape length of P.notoginseng.Among the treatments,ZF significantly increased the single flower weight but reduced the inflorescence diameter,while the effects of BF were opposite to those of ZF;LF significantly increased the stem thickness and reduced the plant height.All treatments significantly increased the seedling rate of three-year-old P.notoginseng,and the increase in the LF group(20.49%)was the largest,followed by those in the ZF(16.80%)and OM(16.40%)groups,and the increase in the BF group(13.08%)was the smallest.Although OM,ZF and BF treatments caused the root weight of individual plants to decrease,the final yield of each treatment was higher than that of the control group,and the increases in the BF and LF groups exceeded 17%(P<0.05).The total saponin outputs of all the treatments except OM were significantly higher than that of the control group.[Conclusions]Under the conditions of this test,the supplementation of organic and medium and trace element fertilizers on the basis of conventional fertilization will help to increase the yield of P.notoginseng.However,the reduction of the total saponin output of P.notoginseng caused by organic fertilizer cannot be ignored.

三七总皂苷对短暂性前脑缺血大鼠海马区神经元的修复作用实验研究

目的:探讨三七总皂苷对短暂性前脑缺血大鼠海马区神经元的修复作用。方法:选择30只SD大鼠,随机分为三七总皂苷组、模型组、假手术组,每组10只。模型组、三七总皂苷组使用四血管闭塞建立短暂性前脑缺血大鼠动物模型。假手术组手术方式同三七总皂苷组,不做卡环夹闭、电灼永久性闭塞,仅将右侧颈总动脉、颈内动脉、颈外动脉暴露,之后逐层缝合。三七总皂苷组造模后灌胃给予50 mg/kg三七总皂苷,每天2次,每次间隔12 h,模型组、假手术组灌胃给予等量的0.5%羟甲基纤维素钠。对比三组干预后7、14、28 d的海马神经细胞凋亡、新生神经元数量,对比三组大鼠的学习记忆能力,对比三组干预后7、14、28 d时测量大鼠脑梗死体积及含水量,对比三组DCX/NeuN染色阳性的细胞数量。结果:模型组干预后7、14、28 d的海马神经细胞凋亡明显较三七总皂苷组、假手术组高(均P<0.05);三七总皂苷组干预后7、14、28 d的海马神经细胞凋亡明显较假手术组高(均P<0.05);三七总皂苷组干预后7、14、28 d的海马新生神经元明显较模型组、假手术组高(均P<0.05);干预后7、14、28 d的模型组新生神经元明显较假手术组高(均P<0.05);三七总皂苷组中,随着干预时间延长,海马神经细胞凋亡明显降低,新生神经元明显升高(均P<0.05)。模型组大鼠的潜伏期、错误次数、第1记忆错误次数、第1天学习错误次数、第5天记忆错误次数、第5天学习错误次数明显较三七总皂苷组、假手术组高(均P<0.05),三七总皂苷组大鼠的潜伏期、错误次数、第1天记忆错误次数、第1天学习错误次数、第5天记忆错误次数、第5天学习错误次数明显较假手术组高(均P<0.05)。三七总皂苷组中,随着干预时间延长,第1天记忆错误次数、第1天学习错误次数、第5天记忆错误次数、第5天学习错误次数明显降低(均P<0.05)。模型组的脑梗死体积、含水量明显较三七总皂苷组、假手术组高(均P<0.05)。三七总皂苷组的脑梗死体积、含水量明显较假手术组高(均P<0.05)。三七总皂苷组中,随着干预时间延长,脑梗死体积、含水量明显降低(均P<0.05)。三七总皂苷组的DCX/NeuN染色阳性细胞数量明显较模型组、假手术组高(均P<0.05),模型组明显较假手术组高(P<0.05)。结论:三七总皂苷可促进短暂性前脑缺血大鼠海马区神经元的修复。

竹节参总皂苷通过HMGB1/TLR4/NF-κB信号通路改善游离脂肪酸诱导肝细胞脂肪变性的实验研究

目的:通过HMGB1/TLR4/NF-κB信号通路探讨竹节参总皂苷(TSPJ)改善游离脂肪酸(FFA)诱导的肝癌细胞HepG2脂肪变性的作用机制。方法:体外培养HepG2细胞,随机分成对照组、FFA组、FFA+高迁移率族蛋白1(HMGB1) p-NC组、FFA+HMGB1 siRNA组、FFA+TSPJ组、FFA+pcDNA+TSPJ组、FFA+HMGB1 pcDNA+TSPJ组、FFA+HMGB1 p-NC+TSPJ组、FFA+HMGB1 siRNA+TSPJ组。RT-qPCR和Western blot检测HMGB1 siRNA干扰效率;油红O染色观察TSPJ对细胞脂质蓄积的影响;检测TSPJ对甘油三酯(TG)的作用。RT-qPCR和Western blot检测各组细胞HMGB1、蛋白Toll样受体4(TLR4)和核因子κB(NF-κB)表达水平。结果:HepG2细胞经0.5 mmol/L FFA处理后,脂质水平及TG含量升高(P<0.05);TSPJ干预使肝细胞脂质水平及TG含量降低(P<0.05);干扰HMGB1表达后,肝细胞脂质水平及TG含量降低(P<0.05);TSPJ处理的同时过表达HMGB1,FFA诱导的脂质水平和TG含量恢复(P<0.05)。FFA诱导使肝细胞HMGB1、TLR4和NF-κB表达升高(P<0.05);TSPJ处理使肝细胞HMGB1、TLR4和NF-κB表达降低(P<0.05);干扰HMGB1表达后TLR4和NF-κB p65进一步降低(P<0.05)。结论:竹节参总皂苷通过HMGB1/TLR4/NF-κB信号通路改善FFA诱导肝细胞脂肪变性。

竹节参总皂苷减轻高脂饮食诱导的小鼠睾丸支持细胞连接功能损伤

目的探究竹节参总皂苷(TSPJ)对高脂饮食诱导的小鼠睾丸支持细胞功能损伤的保护作用及机制。方法将40只C57BL/6J雄性小鼠随机分为正常饮食组、高脂饮食组、TSPJ低剂量(25 mg/kg)组和TSPJ高剂量(75 mg/kg)组,10只/组。正常饮食组给予普通饲料喂养,其余组均给予高脂饲料喂养,TSPJ连续灌胃给药5月,小鼠称重处死,取出睾丸和附睾组织称重并计算其指数;取附睾组织进行精液分析;HE染色观察小鼠睾丸组织形态变化,测量统计生精上皮厚度与生精小管直径;Western blot检测睾丸支持细胞紧密连接功能蛋白ZO-1、Occludin和Claudin11以及外质特化功能蛋白N-cadherin、E-cadherin和β-catenin的表达水平;免疫荧光法检测睾丸组织中ZO-1和β-catenin蛋白定位及表达;免疫荧光双标法检测睾丸支持细胞中LC3B、p-AKT和p-mTOR表达变化。结果与高脂饮食组相比,TSPJ能明显减轻高脂饮食小鼠体质量(P<0.001),增加睾丸和附睾指数(P<0.05,P<0.01),提高精子浓度与精子活率(P<0.05,P<0.01),改善睾丸组织形态异常,增加生精上皮厚度(P<0.05,P<0.001),但对生精小管直径无明显影响。Western blot结果显示,TSPJ能增加ZO-1、Occludin、N-cadherin、E-cadherin和β-catenin的蛋白表达水平(P<0.05,P<0.01,P<0.001),但对Claudin11的表达无显著性影响;免疫荧光结果显示,TSPJ能增加睾丸组织中ZO-1和β-catenin蛋白的表达(P<0.001),下调睾丸支持细胞中LC3B的表达,上调p-AKT和p-mTOR蛋白表达。结论TSPJ能减轻高脂饮食诱导的睾丸支持细胞连接功能损伤,改善小鼠生精功能减退,其机制可能与激活支持细胞AKT/mTOR信号通路抑制自噬有关。

云南不同产地珠子参的品质评价研究

目的:研究不同产地野生珠子参的竹节参皂苷IVa和总皂苷的含量差异,结合农艺性状对三个不同产地珠子参的种质资源进行评价,为珠子参的栽培及优质种质的筛选提供理论基础。方法:以云南三个不同产地野生珠子参为研究对象,采用紫外分光光度法和试剂显色法测定珠子参中总皂苷的含量;采用高效液相色谱法(HPLC)测定珠子参中竹节参皂苷IVa的含量;采用民族植物学田野调查法研究云南三个不同产地珠子参的农艺性状。结果:云南三个不同产地的野生珠子参,以云南省丽江市玉龙县的野生珠子参中总皂苷和竹节参皂苷IVa的含量最高,其珠子参总皂苷的含量为15.21%±1.25%,珠子参中竹节参皂苷IVa的含量为4.78%±0.35%,农艺性状也相对较优。结论:不同产地的珠子参质量差异较大,云南省丽江市玉龙县的野生珠子参品质较好,具有较高的开发利用价值。

Combination of Total Astragalus Extract and Total Panax Notoginseng Saponins Strengthened the Protective Effects on Brain Damage through Improving Energy Metabolism and Inhibiting Apoptosis after Cerebral Ischemia-Reperfusion in Mice

Objective： To explore the effects and molecular mechanisms of the combination between total Astragalus extract （TAE） and total Panax notoginseng saponins （TPNS） against cerebral ischemia- reperfusion injury. Methods： C57BL/6 mice were randomly divided into sham-operated group, model group, TAE （110 mg/kg） group, TPNS （115 mg/kg） group, TAE-TPNS combination group and Edaravone （4 mg/kg） group, treated for 4 days, then, cerebral ischemia-repeffusion injury was established by bilateral common carotid artery （CCA） ligation for 20 min followed by reperfusion for 1 and 24 h. Results： TPNS could increase adenosine triphosphate （ATP） level, TAE and TAE-TPNS combination increased ATP, adenosine diphosphate （ADP） contents and Na＋-K＋-ATPase activity, and the effects of TAE-TPNS combination were stronger than those of TAE or TPNS alone after reperfusion for 1 h. After reperfusion for 24 h, TAE, TPNS and TAE-TPNS combination significantly increased neurocyte survival rate and decreased the apoptosis rate as well as down-regulated the expression of phosphorylated c-June N-terminal kinasel/2 （p-JNK1/2）, cytochrome C （Cyt C）, cysteine aspartic acid-specific protease （Caspase）-9 and Caspase-3. Furthermore, the effects in TAE-TPNS combination were better than those in TAE or TPNS alone. Conclusion： The combination of TAE 110 mg/kg and TPNS 115 mg/kg could strengthen protective effects on cerebral ischemia injury, the mechanism underlying might be related to improving jointly the early energy metabolism, and relieving the delayed apoptosis via inhibiting the mitochondrial apoptosis pathway of JNK signal transduction.

Total Saponins of Panax notoginseng Activate Akt/mTOR Pathway and Exhibit Neuroprotection in vitro and in vivo against Ischemic Damage

Objective:To reveal the neuroprotective effect and the underlying mechanisms of a mixture of the main components of Panax notoginseng saponins(TSPN)on cerebral ischemia-reperfusion injury and oxygenglucose deprivation/reoxygenation(OGD/R)of cultured cortical neurons.Methods:The neuroprotective effect of TSPN was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT)assay,flow cytometry and live/dead cell assays.The morphology of dendrites was detected by immunofluorescence.Middle cerebral artery occlusion(MCAO)was developed in rats as a model of cerebral ischemia-reperfusion.The neuroprotective effect of TSPN was evaluated by neurological scoring,tail suspension test,2,3,5-triphenyltetrazolium chloride(TTC)and Nissl stainings.Western blot analysis,immunohistochemistry and immunofluorescence were used to measure the changes in the Akt/mammalian target of rapamycin(mTOR)signaling pathway.Results:MTT showed that TSPN(50,25 and 12.5μg/m L)protected cortical neurons after OGD/R treatment(P<0.01 or P<0.05).Flow cytometry and live/dead cell assays indicated that 25μg/m L TSPN decreased neuronal apoptosis(P<0.05),and immunofluorescence showed that 25μg/m L TSPN restored the dendritic morphology of damaged neurons(P<0.05).Moreover,12.5μg/m L TSPN downregulated the expression of Beclin-1,Cleaved-caspase 3 and LC3 B-Ⅱ/LC3 B-Ⅰ,and upregulated the levels of phosphorylated(p)-Akt and p-mTOR(P<0.01 or P<0.05).In the MCAO model,50μg/m L TSPN improved defective neurological behavior and reduced infarct volume(P<0.05).Moreover,the expression of Beclin-1 and LC3 B in cerebral ischemic penumbra was downregulated after 50μg/m L TSPN treatment,whereas the p-mTOR level was upregulated(P<0.05 or P<0.01).Conclusions:TSPN promoted neuronal survival and protected dendrite integrity after OGD/R and had a potential therapeutic effect by alleviating neurological deficits and reversing neuronal loss.TSPN promoted p-mTOR and inhibited Beclin-1 to alleviate ischemic damage,which may be the mechanism that underlies the neuroprotective activity of TSPN.

Chemical constituents from acid hydrolyzates of Panax quinquefolius total saponins and their inhibition activity to α-glycosidase and protein tyrosine phosphatase 1B

Objective:To investigate the hypoglycemic components from the acid hydrolyzates of Panax quinquefolius total saponins,and screen the active compounds by in vitro inhibitory activities toα-glycosidase enzymes and protein tyrosine phosphatase-1 B(PTP1 B).Methods:The hydrolyzates were chromatographed repeatedly over silica gel column,and the structures of the compounds were determined by means of NMR.The in vitro bioassay was performed through the inhibitory effects onα-glucosidase or/and PTP1 B.Results:Eight compounds were isolated,which identified as 20(S)-panaxadiol(1),(20 S,24 R)-dammarane-20,24-epoxy-3β,6α,12β,25-tetraol(2),20(R)-dammarane-3β,12β,20,25-tetraol(3),20(S)-dammarane-3β,6α,12β,20,25-pentol(4),20(R)-dammarane-3β,12β,20,25-tetrahydroxy-3β-O-β-D-glucopyranoside(5),β-sitosterol(6),oleanolic acid(7)and 20(S)-protopanaxadiol(8).Compound 5 was ginseng triterpenoid isolated from the acid hydrolysates of total saponins from P.quinquefolius for the first time.In this paper,the possible in vitro inhibitory activities were investigated.Compound 5 exhibited significantly inhibitory activity againstα-glucosidase,and the IC50 value[(0.22±0.21)μmol/L]was about 43-fold lower than positive control.For the PTP1 B inhibition assay,compound 5 indicated the strongest inhibitory effect with IC50 of(5.91±0.38)μmol/L,followed by compound 4 with IC50 of(6.21±0.21)μmol/L,which were all showed competitive inhibitory pattern by using a Lineweaver-Burk plot.Conclusion:These results supported the potential application of dammaranes from acid hydrolyzates of P.quinquefolius total saponins can be used as ingredients of ancillary anti-diabetic agent or functional factor.

三七总皂苷调控SIRT1/FOXO3a/p27通路抑制大鼠肺动脉平滑肌细胞增殖

目的探讨三七总皂苷(PNS)通过SIRT1/FOXO3a/p27通路抑制大鼠肺动脉平滑肌细胞(PASMCs)增殖的作用。方法首先将PASMCs随机分为正常(Control)组和三七总皂苷(PNS)组、野百合碱(MCT)组和野百合碱+三七总皂苷(MCT+PNS)组,CCK8检测PNS对正常细胞的安全浓度,进一步检测PNS抑制增殖的最适浓度;为研究作用机制将PASMCs分为Control组、溶剂(DMSO组)组、MCT组、MCT+PNS组、MCT+PNS+SIRT1抑制剂(MCT+PNS+EX-527)组。造模结束,Edu检测细胞增殖;免疫荧光检测SIRT1、FOXO3a表达;qPCR检测细胞SIRT1、FOXO3a、p27、PCNA的表达。结果CCK8显示:0~400 mg·L^(-1)的PNS对正常细胞无毒(P>0.05),100 mg·L^(-1)的PNS可显著抑制MCT诱导的细胞增殖(P<0.01);Edu显示:MCT组较Control组增殖增多;MCT+PNS组较MCT组增殖减少;MCT+PNS+EX-527组较MCT+PNS组增殖增多。免疫荧光和qPCR显示:MCT组较Control组,SIRT1、FOXO3a、p27表达降低,PCNA表达增高;较MCT组,MCT+PNS组SIRT1、FOXO3a、p27表达增多,PCNA表达降低;较MCT+PNS组,MCT+PNS+EX-527组SIRT1、FOXO3a、p27表达降低,PCNA表达增高。结论PNS可通过SIRT1/FOXO3a/p27通路抑制大鼠PASMCs增殖。

竹节参总皂苷中10种活性成分在正常、肥胖大鼠体内药动学比较

目的比较竹节参总皂苷中人参皂苷Re、Rg1、Rf、Rb1、Rg2、Rd,竹节参皂苷Ⅴ、Ⅳ、Ⅳa,拟人参皂苷RT1在正常、肥胖大鼠体内药动学。方法大鼠随机分为正常组和肥胖组,灌胃给予总皂苷0.4%CMC-Na混悬液(0.8 g/kg),于0、0.08、0.25、0.5、0.75、1、1.5、2、4、6、8、12、24、48 h采血,UPLC-MS法测定各活性成分血药浓度,计算主要药动学参数。结果各活性成分C_(max)、AUC_(0~t)与其在总皂苷中的含量呈正相关。竹节参皂苷Ⅴ、Ⅳ、Ⅳa为体内主要暴露成分,t_(1/2)、MRT_(0~t)更长。与正常组比较,肥胖组各活性成分C_(max)(除人参皂苷Rg1)、AUC_(0~t)(除人参皂苷Re、Rg1和拟人参皂苷RT1)降低,t_(1/2)、MRT_(0~t)、T_(max)无明显变化。结论肥胖可影响竹节参总皂苷中部分活性成分的体内药动学,竹节参皂苷Ⅴ、Ⅳ、Ⅳa可能是竹节参总皂苷改善肥胖的关键组分。

基于UPLC-Q/TOF MS技术探讨竹节参总皂苷对HepG2细胞脂质代谢的改善作用

基于细胞代谢组学技术研究竹节参总皂苷改善HepG2细胞脂质代谢的机制。应用棕榈酸和油酸诱导HepG2细胞脂质沉积模型,竹节参总皂苷干预24 h后进行油红O染色和甘油三酯分析。采用改良的Bligh-Dy⁃er法将细胞内源性代谢物分为极性和非极性部位,运用超高效液相色谱-四极杆飞行时间质谱(UPLC-Q/TOF MS)技术检测代谢物,结合多元统计分析和火山图筛选差异代谢物,并富集代谢通路。结果显示,竹节参总皂苷可显著改善HepG2细胞脂质沉积模型的脂质代谢。从极性部位鉴定了34个差异代谢物,主要为脂肪酸和氨基酸类成分;从非极性部位鉴定了28个差异代谢物,主要为磷脂酰胆碱、磷脂酰乙醇胺、神经酰胺类成分。差异代谢物在竹节参总皂苷干预后均有恢复至正常水平的趋势。该研究提示竹节参总皂苷改善HepG2细胞脂质沉积模型的脂质代谢与调控鞘脂代谢、甘油磷脂代谢、脂肪酸代谢、炎症反应密切相关。

The Effects of Total Saponins of Panax Ginseng on Hematopoietic Progenitor Cells in Healthy Humans and Aplastic Anemia Patients

Ginseng is said to have beneficial effects on anemia. The proliferation effects of totalsaponins of Panax ginseng (TSPG) on hematopoietic progenitor cell in healthy individuals and 29 patientswith aplastic anemia (AA) were observed through bone marrow cultures of burst forming unit-erythroid(BFU-E) , colony forming unit-erythroid (CFU-E) and colony forming unit-granulocyte/macrophage (CFU-GM) in vitrcacompared with methyltestosterone (MT). The results suggest TSPG might prompt the prolif-eration of normal progenitor cellS at a concentration of 20 g/ml. The numbers of BFU-E ,CFU-E and CFU-GM increased by 37. 8±2.9 % , 31. 4±2. 9 % and 33. 3± 4. 0 % respectively over the controls ; further-more TSPG was still useful to BFU-E,CFU-E growth without Epo in vitro, although the colony nurnberswere much lower. Otherwise MT was useless to CFUGM. Of the 29 patients with AA, 14 who respondedto MT showed sensitivity to TSPG in marrow culture (the rising rate of colony formation exceeded 30 % ) ,but immune-mediated AA (patient's peripheral blood mononucleated cell suppressed normalhematopoiesis) and stem cell decreased AA (few of colonies were formed) showed almost no expressionfor TSPG activity because of the immunological suppression system and the absence of progenitors.