人参皂苷Rg1抑制PGF_(2α)诱导心肌细胞肥大

目的观察人参皂苷Rg1(ginsenoside Rg1,Rg1)对前列腺素F2α(prostaglandin F2α,PGF2α)所致心肌肥大的影响。方法利用培养的新生大鼠心肌细胞,以细胞直径、蛋白含量?心房利钠因子(atrial natriuretic factor,ANF)mRNA的表达为心肌肥大标志,观察药物的抗心肌肥大效应。AN-FmRNA的表达用Real-time PCR检测;用Fura-2/AM负载的培养心肌细胞检测细胞内游离钙浓度([Ca2+]i);一氧化氮试剂盒测定细胞上清液中一氧化氮(nitric oxide,NO)代谢物的含量,以探讨Rg1可能的作用机制。结果PGF2α0.1μmol·L-1使心肌细胞直径明显增大,蛋白质含量明显增加,ANF mRNA的表达增强,并使[Ca2+]i明显升高。Rg115.6、31.2和62.4μmol·L-1使经PGF2α处理的心肌细胞的直径分别缩短18.4%、32.7%和43.8%;使心肌细胞蛋白含量分别减少8.2%、14.6%和17.7%;Rg1 62.4μmol·L-1还使ANF mRNA的表达降低;Rg1 15.6、31.2和62.4μmol·L-1呈剂量依赖地抑制PGF2α所致的[Ca2+]i升高;NO前体L-精氨酸(L-arg)1 mmol.L-1具有相似的作用。此外,NO合酶抑制剂L-NAME可取消L-arg的作用,并部分抑制Rg1的效应;Rg1还明显升高心肌细胞上清液NO代谢物的含量。结论Rg1可抑制PGF2α诱导的心肌细胞肥大,该作用可能与其促进心肌细胞NO的释放,降低由PGF2α所升高的心肌细胞[Ca2+]i有关。     

人参皂苷Rg3诱导膀胱癌细胞系EJ的凋亡作用

目的：探讨Rg3抑制膀胱癌细胞的作用及其诱导凋亡的机制。方法：用一定质量浓度的Rg3 处理膀胱癌细胞系EJ细胞,MTT法检测细胞增殖活力和抑制率50%的时候药物的浓度(IC₅₀)；Hoechst33258荧光染色观察凋亡细胞的形态；流式细胞仪分析细胞周期及细胞凋亡率；免疫细胞化学观察caspase-3的表达变化；琼脂糖凝胶电泳测定DNA梯状带。结果：Rg3抑制EJ细胞生长,呈浓度依赖关系,Rg3处理细胞48 h的IC₅₀为125.5 mg·L^-1；经150 mg·L^-1Rg3处理细胞24 h和48 h后,观察到典型的凋亡细胞形态,即染色质凝集、核片段化、凋亡小体、核内致密的颗粒状荧光及caspase-3的表达增加；并呈时效依赖关系；用75 mg·L^-1Rg3处理细胞24,48,150 mg·L^-1的Rg3处理细胞48 h后,Rg3诱导了细胞凋亡和调节细胞周期,S期和G₂/M期的细胞比率增加,G₀/G₁的细胞比率下降；凋亡率从对照组的(1.05±0.17)%分别上升到(8.41±0.98)%,(18.57±2.20)% 和(33.98±1.64)%,琼脂糖凝胶电泳检测出典型的DNA梯形条带,其效应随药物浓度的增加和作用时间的延长而增强。结论：Rg3可通过诱导EJ细胞凋亡而发挥其抑制细胞增殖的作用。     

心脑治胶囊中人参皂苷Rg_1、Rb_1及三七皂苷R_1的检测方法

目的建立心脑治胶囊中人参皂苷及三七皂苷含量的检测方法。方法采用高效液相色谱法,色谱柱为ZorbaxC18柱,乙腈-水梯度洗脱(0~3min,20%~25%乙腈;3~15min,25%~45%乙腈);流速1.0mL.min-1;检测波长203nm。结果人参皂苷Rg1、人参皂苷Rb1、三七皂苷R1线性关系良好(r≥0.999),平均回收率(n=5)分别为105.68%(RSD=1.52%)、104.34%(RSD=0.73%)、102.93%(RSD=1.26%)。结论本含量测定方法灵敏、简便、快速、准确,适用于心脑治胶囊内在质量的控制。     

Neuroprotective effects of ginseng total saponin and ginsenosides Rb1 and Rg1 on spinal cord neurons in vitro

Spinal cord injury is a major cause of disability and results in many serious physical, psychological, and social difficulties. Numerous studies have shown that traumatic spinal cord injuries (SCI) lead to neuronal loss and axonal degeneration in and around the injury site that cause partial disability or complete paralysis. An important strategy in the treatment of SCI is to promote neuron survival and axon outgrowth, making possible the recovery of neural connections. Using an in vitro survival assay, we have identified ginsenosides Rb1 and Rg1, extracted from ginseng root (Panax ginseng C. A. Meyer), as efficient neuroprotective agents for spinal cord neurons. These compounds protect spinal neurons from excitotoxicity induced by glutamate and kainic acid, as well as oxidative stress induced by H(2)O(2). The neuroprotective effects are dose-dependent. The optimal doses are 20-40 microM for ginsenosides Rb1 and Rg1. The effects are specific for Rb1 and Rg1, since a third ginsenoside, Re, did not exhibit any activity. Ginseng has been used for thousands of years in the treatment of neurological disorders and other diseases in Asia. Ginsenosides Rb1 and Rg1 represent potentially effective therapeutic agents for spinal cord injuries.     

人参皂甙Rb1、黄芪抗大鼠大脑皮层神经细胞缺氧性凋亡研究

目的研究人参皂甙Rb1、黄芪对体外培养的新生大鼠大脑皮层神经细胞缺氧性凋亡的保护机理。方法应用Neurobasal加B27 Supplement体外培养大鼠大脑皮层神经细胞;流式细胞仪检测神经细胞凋亡率;Hoechst33342荧光染色法检测神经细胞凋亡率。结果(1)流式细胞仪测定结果:正常对照组神经细胞凋亡率为0.78%,凋亡阳性组细胞凋亡率为31.61%,人参皂甙Rb1 1、2、3组细胞凋亡率分别为8.52%、4.86%、1.43%,黄芪注射液1、2、3组细胞凋亡率分别为16.06%、7.03%、1.22%;(2)神经细胞凋亡阳性组可见有明显的凋亡梯状条带,而正常对照组、人参皂甙Rb1 2组、黄芪注射液2组未见有明显的凋亡梯状条带;(3)Hoechst 33342荧光染色结果:正常对照组神经细胞凋亡率为(8.24±0.31)%,凋亡阳性组细胞凋亡率为(41.30±2.56)%;与凋亡阳性组比较,缺氧前加BDNF组细胞凋亡率为(21.06±0.72)%(P<0.05),缺氧后加BDNF组细胞凋亡率为(27.82±3.65)%(P<0.01);缺氧前加人参皂甙Rb1 3组细胞凋亡率为(12.57±1.61)%(P<0.05),缺氧后加人参皂甙Rb1 4组细胞凋亡率为(23.71±2.24)%(P<0.01);缺氧前加黄芪注射液1、2、3组细胞凋亡率分别为(20.28±1.00)%、(18.67±0.57)%、(13.84±0.79)%(P<0.05),缺氧后加黄芪注射液4组细胞凋亡率为(26.46±2.18)%(P<0.01)。结论人参皂甙Rb1在100μg/mL,黄芪在10~100 mg/mL的浓度范围內,具有抗缺氧的神经细胞凋亡的作用。     

人参皂苷抗高脂血症及动脉粥样硬化的实验研究

目的:探讨人参皂苷防治高脂血症、动脉粥样硬化的疗效与可能的机制。方法:给C57/BL-ApoE基因敲除小鼠喂饲胆固醇饲料90天,形成实验性高胆固醇血症和动脉粥样硬化斑块,同时中药治疗组连续给人参皂苷喂养90天,实验结束后测量小鼠血脂,观察动脉壁的光镜改变。结果显示:人参皂苷能抑制血清总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)的上升。结论:说明人参皂苷能治疗实验性高脂血症,抑制动脉粥样硬化斑块的形成。     

Promotion of hair growth by ginseng radix on cultured mouse vibrissal hair follicles

A 70% methanol extract from red ginseng (steamed and dried roots of Panax ginseng C. A. Meyer, a kind of Ginseng Radix) had superior activity to that of white ginseng (peeled and dried root of P. ginseng, another kind of Ginseng Radix) in a hair growth promoting assay using mouse vibrissal follicles in organ culture. Of the major constituents of P. ginseng, ginsenoside-Rb(1) (G-Rb(1)) exhibited activity, but ginsenoside-Rg(1) (G-Rg(1)) and -Ro (G-Ro) were ineffective. Additionally, 20(S)-ginsenoside-Rg(3) (20(S)-G-Rg(3)) formed by the processing of red ginseng from the crude root of P. ginseng also showed hair growth promoting activity. These results indicate that Ginseng Radix possesses hair growth promoting activity, and its bioactive components are partially attributable to the ginseng saponin components mentioned above.     

长寿液质量标准研究

目的　制定长寿液质量标准。方法　采用了LC法对处方中的枸杞子、当归和川芎等3种药材追行定性鉴别；采用HPLC法对处方中人参的人参皂苷RgI追行了含量测定，色谱条件：依利特Hypersil　C18色谱柱（4．6×150mm，5μm）；流动相乙腈-0．1％磷酸溶液（19．8：70．2）；流速1．0ml／min；检测波长为203nm。结果　人参皂苷Rgl在0．0298125—0．477mg／ml范围内线性关系良好，r=1．0000，平均回收率为101．50％，RSD为0．61％。结论　本法操作简便，能准确、可靠地追行定性、定量检测，可有效控制长寿液的质量。