GPCR-Gs mediates the protective effects of ginsenoside Rb1 against oxygen-glucose deprivation/re-oxygenation-induced astrocyte injury

Objectives:To investigate whether the protective actions of ginsenoside Rb1(Rb1)on astrocytes are mediated through the G_(s)-type G-protein-coupled receptor(GPCR-G_(s)).Methods:Primary astrocyte cultures derived from neonatal mouse brain were used.Astrocyte injury was induced via oxygen-glucose deprivation/re-oxygenation(OGD/R).Cell morphology,viability,lactate dehydrogenase(LDH)leakage,apoptosis,glutamate uptake,and brain-derived neurotrophic factor(BDNF)secretion were assessed to gauge cell survival and functionality.Western blot was used to investigate the cyclic adenosine monophosphate(cAMP)and protein kinase B(Akt)signaling pathways.GPCR-G_(s)-specific inhibitors and molecular docking were used to identify target receptors.Results:Rb1 at concentrations ranging from 0.8 to 5μM did not significantly affect the viability,glutamate uptake,or BDNF secretion in normal astrocytes.OGD/R reduced astrocyte viability,increasing their LDH leakage and apoptosis rate.It also decreased glutamate uptake and BDNF secretion by these cells.Rb1 had protective effects of astrocytes challenged by OGD/R,by improving viability,reducing apoptosis,and enhancing glutamate uptake and BDNF secretion.Additionally,Rb1 activated the cAMP and Akt pathways in these cells.When the GPCR-G_(s) inhibitor NF449 was introduced,the protective effects of Rb1 completely disappeared,and its activation of cAMP and Akt signaling pathways was significantly inhibited.Conclusion:Rb1 protects against astrocytes from OGD/R-induced injury through GPCR-G_(s) mediation.

Effect of ginsenoside Rg1 on hematopoietic stem cells in treating aplastic anemia in mice via MAPK pathway

BACKGROUND Aplastic anemia(AA)presents a significant clinical challenge as a life-threatening condition due to failure to produce essential blood cells,with the current the-rapeutic options being notably limited.AIM To assess the therapeutic potential of ginsenoside Rg1 on AA,specifically its protective effects,while elucidating the mechanism at play.METHODS We employed a model of myelosuppression induced by cyclophosphamide(CTX)in C57 mice,followed by administration of ginsenoside Rg1 over 13 d.The invest-igation included examining the bone marrow,thymus and spleen for pathological changes via hematoxylin-eosin staining.Moreover,orbital blood of mice was collected for blood routine examinations.Flow cytometry was employed to identify the impact of ginsenoside Rg1 on cell apoptosis and cycle in the bone marrow of AA mice.Additionally,the study further evaluated cytokine levels with enzyme-linked immunosorbent assay and analyzed the expression of key proteins in the MAPK signaling pathway via western blot.RESULTS Administration of CTX led to significant damage to the bone marrow’s structural integrity and a reduction in hematopoietic cells,establishing a model of AA.Ginsenoside Rg1 successfully reversed hematopoietic dysfunction in AA mice.In comparison to the AA group,ginsenoside Rg1 provided relief by reducing the induction of cell apoptosis and inflammation factors caused by CTX.Furthermore,it helped alleviate the blockade in the cell cycle.Treatment with ginsenoside Rg1 significantly alleviated myelosuppression in mice by inhibiting the MAPK signaling pathway.CONCLUSION This study suggested that ginsenoside Rg1 addresses AA by alleviating myelosuppression,primarily through modulating the MAPK signaling pathway,which paves the way for a novel therapeutic strategy in treating AA,highlighting the potential of ginsenoside Rg1 as a beneficial intervention.

人参皂苷Rb_(1)治疗非酒精性脂肪性肝病药理作用的研究进展

非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)已成为全球最常见的慢性肝病,主要表现为肝脏中脂肪过度沉积,并可进一步导致肝纤维化,以及肝硬化和肝细胞癌等终末期肝病。NAFLD发生和发展的分子机制尚不完全清楚,目前还没有批准的药物用于治疗NAFLD。人参皂苷Rb_(1)是人参中最丰富和最具生物活性的分子,具有调节糖脂代谢、抗炎、抗氧化和神经保护等多种药理活性。就NAFLD发病机制及人参皂苷Rb_(1)治疗NAFLD药理作用进行简要综述,为今后人参皂苷Rb_(1)的研究和临床治疗提供参考。

Effect of Ginsenoside-Rb_1 on Cardiomyocyte Apoptosis after Ischemia and Reper fusion in Rats

The effect of ginsenoside Rb 1 on cardiomyocyte apotosis after ischemia (30 min ) and reperfusion (6 h) in rats was observed. The ischemia/ reperfusion heart mo del was established by ligating left anterior descending branch of coronary arte ry in Wistar rats. The apoptotic cardiomyocytes were examined under transmission electron microscopy and counted by in situ nick end labeling (TUNEL) method and light microscopy. Results showed that (1) The apoptotic cardiomyocytes were found in ischemic regions in the ischemia/reperfusion group, but not in the sh am oper ating group under transmission electron microscopy; (2) The number of apoptotic cells were 134.45±45.61/field in the ischemia/reperfusion group, 0/field in the sham operating group and 51.65±13.71/field in the ginsenoside Rb 1 treated group. The differences were significant among the three groups ( P <0.01). It was concluded that myocardial ischemia reperfusion could induce cardiomyocyte a poptosis, and ginsenoside Rb 1 could significantly inhibit cardiomyocyte apopto sis induced by ischemia reperfusion in rats, indicating that ginsenoside Rb 1 could inhibit cardiomyocyte apoptosis induced by ischemia reperfusion, thus alleviating ischemia reperfusion injury.

Ginsenoside Rg1 protects against ischemia-induced neuron damage by regulating the rno-miRNA-27a-3p/PPARγaxis

A preliminary miRNA screening showed that expression levels of rno-miRNA-27a-3p were significantly increased in the serum and brain tissues of rats undergoing cerebral ischemia.In recent years,there is evidence of the protective capacity of the saponins extracted from panax ginseng and its primary active ingredient ginsenosideRg1oncerebral ischemic injury.Methods:Fetal rat neurons(FRNs)were cultured in glucose-and-serumfree medium and exposed to hypoxia to establish a cerebral ischemia model in vitro(oxygen and glucose deprivation model,OGD).Antioxidant indexes(CAT,SOD),inflammatory markers(MPO,TNF-αand IL-6),and the expression of apoptosis and proliferation associated proteins(NF kB-p65,Caspase 3-cleaved,BCL-2)were examined.Results:Pre-treatment of Rg1(30–100μg/mL)could effectively inhibit the decline of antioxidant indexes(CAT,SOD)and increase in inflammatory markers(MPO,TNF-αand IL-6),and effectively inhibited the apoptosis in FRNs induced by OGD in a gradient-dependent manner.The mechanism analysis showed that the role of Rg1 in protecting against ischemia-induced neuron damage depends on its indirect up-regulation of PPAR protein via suppression of rnomiRNA-27a-3p.Moreover,these effects of Rg1 could be reversed by exogenous rno-miRNA-27a-3p and PPAR gene silencing in FRNs exposed to OGD.Conclusion:To summarize,our study demonstrates that Rg1 could effectively attenuate neuronal damage caused by cerebral ischemia via the rno-miRNA-27a-3p/PPARγpathway.Further,clarification of the novel mechanism will certainly improve our previous understanding of the role of Rg1 and enhancing its level in treatments for alleviating ischemic brain injury.

Ginsenoside F1 administration promotes UCP1-dependent fat browning and ameliorates obesity-associated insulin resistance

Obesity-induced type 2 diabetes is mainly due to excessive free fatty acids leading to insulin resistance.Increasing thermogenesis is regarded as an effective strategy for hypolipidemia and hypoglycemia.Ginsenoside is a natural active component in Panax ginseng C.A.Meyer,and some of them enhance thermogenesis.However,there are few studies on the mechanism and target of ginsenosides enhancing thermogenesis.Using thermogenic protein uncoupling protein 1(UCP1)-luciferase reporter assay,we identifi ed ginsenoside F1 as a novel UCP1 activator in the ginsenosides library.Using pull down assay and inhibitor interference,we found F1 binds toβ3-adrenergic receptors(β3-AR)to enhance UCP1 expression via cAMP/PKA/CREB pathway.We also investigated the ability of F1 on energy metabolism in obesity-induced diabetic mice,including body weight,body composition and energy expenditure.The results of proteomics showed that F1 signifi cantly up-regulated thermogenesis proteins and lipolytic proteins,but down-regulated fatty acid synthesis proteins.Ginsenoside F1 increased thermogenesis and ameliorated insulin resistance specifi cally by promoting the browning of white adipose tissue in obese mice.Additionally,ginsenoside F1 improves norepinephrine-induced insulin resistance in adipocytes and hepatocytes,and shows a stronger mitochondria respiration ability than norepinephrine.These fi ndings suggest that ginsenoside F1 is a promising lead compound in the improvement of insulin resistance.

Protective effect of ginsenoside Rg1 on 661W cells exposed to oxygen-glucose deprivation/reperfusion via keap1/nrf2 pathway

AIM:To construct an in vitro model of oxygen-glucose deprivation/reperfusion(OGD/R)induced injury to the optic nerve and to study the oxidative damage mechanism of ischemia-reperfusion(I/R)injury in 661W cells and the protective effect of ginsenoside Rg1.METHODS:The 661W cells were treated with different concentrations of Na2S2O4 to establish OGD/R model in vitro.Apoptosis,intracellular reactive oxygen species(ROS)levels and superoxide dismutase(SOD)levels were measured at different time points during the reperfusion injury process.The injury model was pretreated with graded concentrations of ginsenoside Rg1.Real-time polymerase chain reaction(PCR)was used to measure the expression levels of cytochrome C(cyt C)/B-cell lymphoma-2(Bcl2)/Bcl2 associated protein X(Bax),heme oxygenase-1(HO-1),caspase9,nuclear factor erythroid 2-related factor 2(nrf2),kelch-like ECH-associated protein 1(keap1)and other genes.Western blot was used to detect the expression of nrf2,phosphorylated nrf2(pnrf2)and keap1 protein levels.RESULTS:Compared to the untreated group,the cell activity of 661W cells treated with Na2S2O4 for 6 and 8h decreased(P<0.01).Additionally,the ROS content increased and SOD levels decreased significantly(P<0.01).In contrast,treatment with ginsenoside Rg1 reversed the cell viability and SOD levels in comparison to the Na_(2)S_(2)O_(4)treated group(P<0.01).Moreover,Rg1 reduced the levels of caspase3,caspase9,and cyt C,while increasing the Bcl2/Bax level.These differences were all statistically significant(P<0.05).Western blot analysis showed no significant difference in the protein expression levels of keap1 and nrf2 with Rg1 treatment,however,Rg1 significantly increased the ratio of pnrf2/nrf2 protein expression compared to the Na_(2)S_(2)O_(4)treated group(P<0.001).CONCLUSION:The OGD/R process is induced in 661W cells using Na_(2)S_(2)O_(4).Rg1 inhibits OGD/R-induced oxidative damage and alleviates the extent of apoptosis in 661W cells through the keap1/nrf2 pathway.These results suggest a potential protective effect of Rg1 against retinal I/R injury.

Ginsenoside Rb1 improves energy metabolism after spinal cord injury

Mitochondrial damage caused by oxidative stress and energy deficiency induced by focal ischemia and hypoxia are important factors that aggravate diseases.Studies have shown that ginsenoside Rb1 has neurotrophic and neuroprotective effects.However,whether it influences energy metabolism after spinal cord injury remains unclear.In this study,we treated mouse and cell models of spinal cord injury with ginsenoside Rb1.We found that ginsenoside Rb1 remarkably inhibited neuronal oxidative stress,protected mitochondria,promoted neuronal metabolic reprogramming,increased glycolytic activity and ATP production,and promoted the survival of motor neurons in the anterior horn and the recovery of motor function in the hind limb.Because sirtuin 3 regulates glycolysis and oxidative stress,mouse and cell models of spinal cord injury were treated with the sirtuin 3 inhibitor 3-TYP.When Sirt3 expression was suppressed,we found that the therapeutic effects of ginsenoside Rb1 on spinal cord injury were remarkably inhibited.Therefore,ginsenoside Rb1 is considered a potential drug for the treatment of spinal cord injury,and its therapeutic effects are closely related to sirtuin 3.

高效液相色谱法同时测定三七总皂苷中人参皂苷Rg_1、Re、Rb_1与三七皂苷R_1含量

目的建立高效液相色谱法同时测定三七总皂苷中人参皂苷Rg1、Re、Rb1与三七皂苷R1的方法。方法采用高效液相色谱法 ,固定相为氨基键合相 ,流动相为乙腈 水 ( 81∶1 9,V∶V) ,检测波长2 0 3nm。结果三七总皂苷中人参皂苷Rg1、Re、Rb1和三七皂苷R1与其他成分分离良好 ,保留时间分别约为 5 7min、8 9min、2 5 1min和 9 9min。人参皂苷Rg1在 80～ 2 80mg/L(r =0 9992 )、Re在 2 0～ 1 80mg/L(r=0 9993 )、Rb1在 95～ 2 85mg/L(r=0 9991 )、三七皂苷R1在1 8～ 1 4 6mg/L(r=0 9991 )内线性关系良好 ,人参皂苷Rg1、Re、Rb1和三七皂苷R1的回收率分别为 99 1 %、98 4 %、98 6%和 97 1 % ,RSD分别为 2 1 %、2 0 %、2 2 %、2 8%。

人参皂苷Rb_1和Rd对不同类型记忆障碍模型小鼠学习记忆功能的影响

分别以东莨菪碱 ,环己米特和乙醇致小鼠获得性 ,巩固性和再现性记忆障碍 ,用跳台实验观察人参皂苷Rb1和Rd对学习记忆功能的影响 .结果表明 ,人参皂苷Rb1(10 0 ,5 0mg·kg- 1,ig)和人参皂苷Rd(2 0 ,10mg·kg- 1,ip)每天上午 9:0 0给药 1次 ,连续给药 7d ,对东莨菪碱和环己米特所致的小鼠获得性及巩固性记忆障碍均具有明显的改善作用 ,而对乙醇所致的小鼠记忆再现障碍则无明显影响 .结果表明 ,人参皂苷Rb1(ig)和Rd(ip)对小鼠学习记忆功能有增强作用 .

人参皂甙Rb_1与Re对大鼠缺血再灌注心肌细胞凋亡的影响

目的 观察人参皂甙Rb1与Re对缺血再灌注心肌细胞凋亡的影响 ,并比较两者的效应差异。方法 结扎Wistar大鼠左冠状动脉前降支 ,建立大鼠缺血再灌注动物模型 ;采用透射电镜、缺口末端标记法检测心肌凋亡细胞 ,利用光学显微镜进行细胞计数。结果 (1)透射电镜发现缺血再灌注组缺血区出现心肌凋亡细胞 ,假手术组未发现心肌凋亡细胞 ;(2 )缺血再灌注组心肌细胞凋亡数为 134 45± 45 6 1个 /视野 ,人参皂甙Rb1治疗组 5 1 6 5± 13 71个 /视野 ,人参皂甙Re治疗组 90 6 6± 19 2 2个 /视野 ,三组间有非常显著性差异 (P <0 0 1)。结论 心肌缺血再灌注诱导心肌细胞凋亡 ,人参皂甙Rb1和Re均可显著减少缺血再灌注心肌细胞的凋亡。证实人参皂甙Rb1与Re均有抑制缺血再灌注心肌细胞凋亡 ,减轻心肌缺血再灌注损伤的作用 ;人参皂甙Rb1的抗心肌细胞凋亡作用较Re的效果为佳。

HPLC法同时测定三七总皂苷中人参皂苷Rg_1与Rb_1的含量

目的 建立 HPL C同时测定三七总皂苷中人参皂苷 Rg1 与 Rb1 的方法。方法 采用 HPL C法 ,以茶碱为内标 ,氨基键合相为固定相 ,流动相为乙腈 -水 (80∶ 2 0 ) ,检测波长 2 0 3nm。结果 三七总皂苷中人参皂苷 Rg1 和 Rb1与其他成分分离良好 ,保留时间分别约为 5 .7和 2 1.5 min。人参皂苷 Rg1 在 80～ 2 80μg/ m L (r=0 .9995 ) ,Rb1 在80～ 2 4 0 μg/ m L(r=0 .9993)线性关系良好 ,Rg1 和 Rb1 加样回收率分别为 97.1%和 98.4 %,RSD分别为 2 .4 4 %和 2 .35 %(n=9)。结论 本法操作简便 ,准确 ,重现性好 ,可用于人参及三七的质量控制。

真菌对人参皂苷Rb_1及人参二醇系皂苷的代谢作用

目的 研究真菌对人参皂苷Rb1 (G Rb1 )及人参二醇系皂苷 (PDS)的代谢作用。方法 在恒温振荡条件下 ,10种真菌对其底物G Rb1 及PDS分别进行代谢 ,不同时间采集代谢物 ,正丁醇萃取 ,用薄层色谱 (TLC)、高效液相色谱 (HPLC)和电喷雾质谱 (ESI MS)检测代谢成分。结果 代谢产物中存在化合物G Rb1 ,G Rd ,G F2 ,CK和Ppd。结论 发现 6种真菌对底物有不同程度的代谢作用 ,其代谢过程可能为G Rb1 (或PDS)→G Rd→G F2 →CK→Ppd ,并通过控制这一过程可获得目的代谢产物CK。

人参皂甙Rb_1促进大鼠雪旺细胞增殖的实验研究

目的 观察人参皂甙 Rb1 对体外培养的大鼠坐骨神经雪旺细胞增殖能力的影响 ,探讨其促进神经再生的作用机制。 方法 取 SD雄性大鼠坐骨神经体外培养第 2代第 5天 ,加入不同浓度的人参皂甙 Rb1 ,利用 MTT比色分析法、3H-胸腺嘧啶核甙测定法检测不同浓度人参皂甙 Rb1 在不同培养时间对体外培养大鼠雪旺细胞增殖的影响。 结果 人参皂甙 Rb1 在 10μg/ ml的浓度对雪旺细胞增殖有明显促进作用 ,高浓度的人参皂甙 Rb1 1mg/ ml则显示抑制作用。而 2 0 0μg/ ml人参皂甙 Rb1 对细胞增殖的促进作用与对照组相近。 结论 人参皂甙 Rb1 在适当浓度范围内可以促进雪旺细胞的增殖 ,从而为促进活体神经损伤的修复途径提供了一些研究基础。

超高速液相测定西洋参中人参皂苷Rg_1、Re、Rb_1的含量研究

目的:建立测定西洋参中人参皂苷Rg_1、Re、Rb_1含量的超高速液相色谱法。方法:采用Waters Acquity UPLC BEHC_(18)(100 mm×2.1 mm,1.7μm)柱分离,以甲醇-水为流动相进行梯度洗脱,流速为0.2ml·min^(-1),柱温为35℃,在203 nm处检测。结果:人参皂苷Rg_1、Re、Rb_1在测定范围内有良好的线性关系,其平均回收率为97.36%～99.23%,RSD为1.16%～1.39%(n=6)。结论:本法操作简便,准确度高,重复性好,可作为西洋参质量控制方法之一。

RP-HPLC梯度法测定血塞通滴丸中三七皂苷R_1、人参皂苷Rb_1及人参皂苷Rg_1的含量

目的 :用高效液相色谱梯度洗脱法同时检测血塞通滴丸中三七皂苷R1、人参皂苷Rb1、人参皂苷Rg13种皂苷含量的方法 ,为制定质量标准中含量测定方法及含量限度提供了依据。方法 :用大连SpherisorbC18分析柱 ,乙腈 水线性梯度洗脱 ,0～ 15min(2 0∶80 - 4 0∶6 0 ) ,流速 1.0mL·min-1,检测波长 2 0 3nm。结果 :R1,Rf1和Rb1线性范围分别为 1.0 2～ 9.18μg ,4 .8～ 4 3.4 μg和 4 .6～ 4 1.6 μg。该方法回收率R1为 10 1.0 % (RSD =3.18% ) ,Rb1为 10 0 .0 % (RSD =1.19% ) ,Rg1为 99.5 % (RSD =3.0 2 % )。结论 :HPLC梯度洗脱法能将多种皂苷很好地分离检测 ,提高了时效 ,减少了误差 ,结果表明该方法准确可靠 ,重现性好 。

HPLC-ELSD测定三七胶囊中人参皂苷Rg_1和Rb_1

目的 :研究三七胶囊的质量标准。方法 :应用HPLC ELSD对三七胶囊中的活性成分人参皂苷Rg1和Rb1的含量进行测定。结果 :人参皂苷Rg1在 1.3 μg～ 6.5μg范围内呈现良好的线性关系 (r =0 .9994) ,平均回收率为 98.3 0 % ,RSD为 1.2 0 % (n =5)。人参皂苷Rb1在 1.0 μg～ 5.0 μg范围内呈现良好的线性关系 (r =0 .9996) ,平均回收率为 97.12 % ,RSD为 1.17% (n =5)。结论 :该方法简便 ,快速 ,重现性好 。

人参皂甙Rb_1、Rg_1、Re和Rh_1对HeLa细胞的影响

本研究应用细胞化学及ＭＴＴ法，测定人宫颈癌细胞（ＨｅＬａ）在人参皂甙Ｒｂ１、Ｒｇ１、Ｒｅ、Ｒｈ１作用３～５ｄ后，细胞增殖和细胞化学含量的变化。结果表明，四种人参单体皂甙与人参根总皂甙作用相似，可以抑制ＨｅＬａ细胞的增殖，降低ＨｅＬａ细胞内多糖（ＰＡＳ）、葡萄糖６磷酸脱氢酶（Ｇ６ＰＤＨ）、葡萄糖６磷酸酶（Ｇ６Ｐａｓｅ）、乳酸脱氢酶（ＬＤＨ）和琥珀酸脱氢酶（ＳＤＨ）的含量。本文提示，不同的人参单体皂甙均可以降低癌细胞的增殖和代谢活性，但Ｒｅ、Ｒｈ１的作用更显著。

人参皂甙Rb_1对大鼠缺血再灌注心肌细胞Bcl-2、Bax、Bad、Fas基因表达的影响

目的 观察人参皂甙Rb1对缺血再灌注心肌细胞Bcl 2、Bax、Bad、Fas基因表达的影响。方法 结扎 /松解Wistar大鼠左冠状动脉前降支 ,建立大鼠缺血再灌注动物模型 ,免疫组化法检测Bcl 2、Bax、Bad、Fas基因的蛋白表达 ,并利用图象分析系统测量蛋白阳性表达区域平均光密度值 ,进行定量分析。结果 缺血再灌注组及Rb1治疗组Bcl 2、Bax、Bad、Fas基因的表达较假手术组明显增加 (P <0 0 5 ) ,Rb1治疗组Bcl 2的表达与缺血再灌注组比较无明显差异 (P >0 0 5 ) ,而Bax、Bad、Fas的表达明显下降 (P <0 0 5 ) ,人参皂甙Rb1治疗组Bcl 2 /Bax、Bcl 2 /Bad以及Bcl 2 /Fas比值均较假手术组与缺血再灌注组明显增加。结论 人参皂甙Rb1治疗可以抑制缺血再灌注心肌细胞中促凋亡基因Bax、Bad、Fas的表达 ,并使Bcl 2 /Bax、Bcl 2 /Bad以及Bcl 2 /Fas比值增加。

人参皂甙Rb_1对应激性性行为缺损的保护作用及机制

用悬吊应激模型，小鼠连续应激小鼠１０ｄ，每天应激一次，并循序增加应激强度，每次应激前３０ｍｉｎ，ｉｐ人参皂甙Ｒｂ１，观察人参皂甙Ｒｂ１对应激性性行为低下的保护作用。结果表明，应激模型组小鼠性行为明显减少，甚至达到缺损的程度，同时血浆睾酮水平明显降低，而Ｒｂ１各剂量组（２５，５，１０ｍｇ·ｋｇ－１）对应激引起的性行为低下及血浆睾酮水平下降均有明显的对抗作用。提示人参皂甙Ｒｂ１对应激性性行为低下有保护作用，其机制可能与Ｒｂ１抑制应激动物血浆皮质酮升高和提高睾酮水平有关。