A Cocktail of Natural Compounds Holds Promise for New Immunotherapeutic Potential in Head and Neck Cancer / 中国结合医学杂志

OBJECTIVE@#To obtain detailed understanding on the gene regulation of natural compounds in altering prognosis of head and neck squamous cell carcinomas (HNSC).@*METHODS@#Gene expression data of HNSC samples and peripheral blood mononuclear cells (PBMCs) of HNSC patients were collected from Gene Expression Omnibus (GEO). Differential gene expression analysis of GEO datasets were achieved by the GEO2R tool. Common differentially expressed gerres (DEGs) were screened by comparing DEGs of HNSC with those of PBMCs. The combination was further analyzed for regulating pathways and biological processes that were affected.@*RESULTS@#Totally 110 DEGs were retrieved and identified to be involved in biological processes related to tumor regulation. Then 102 natural compounds were screened for a combination such that the expression of all 110 commonly DEGs was altered. A combination of salidroside, ginsenoside Rd, oridonin, britanin, and scutellarein was chosen. A multifaceted, multi-dimensional tumor regression was showed by altering autophagy, apoptosis, inhibiting cell proliferation, angiogenesis, metastasis and inflammatory cytokines production.@*CONCLUSIONS@#This study has helped develop a unique combination of natural compounds that will markedly reduce the propensity of development of drug resistance in tumors and immune evasion by tumors. The result is crucial to developing a combinatorial natural therapeutic cocktail with accentuated immunotherapeutic potential.

Quality evaluation of ginsenoside reference substances based on qNMR spectroscopy / 中国中药杂志

The present study established a quality evaluation method for ginsenoside reference substances based on quantitative nuclear magnetic resonance(qNMR) spectroscopy. ~1H-NMR spectra were collected on Bruker Avance Ⅲ 500 MHz NMR spectrometer equipped with a 5 mm BBO probe. The acquire parameters were set up as follows: pulse sequence of 30°, D_1=20 s, probe temperature= 303 K, and the scan number = 32. Dimethyl terephthalate, a high-quality ~1H-qNMR standard, was used as the internal standard and measured by the absolute quantitative method. Methyl peaks of comparatively good sensitivity were selected for quantification, and linear fitting deconvolution was adopted to improve the accuracy of integration results. The qNMR spectroscopy-based method was established and validated, which was then used for the quality evaluation of ginsenoside Rg_1, ginsenoside Re, ginsenoside Rb_1, ginsenoside Rd, and notoginsenoside R_1. The results suggested that the content of these ginsenoside reference standards obtained from the qNMR spectroscopy-based method was lower than that detected by the normalization method in HPLC provided by the manufacturers. In conclusion, the qNMR spectroscopy-based method can ensure the quality of ginsenoside reference substances and provide powerful support for the accurate quality evaluation of Chinese medicine and its preparations. The qNMR spectroscopy-based method is simple, rapid, and accurate, which can be developed for the quantitative assay of Chinese medicine standard references.

Ginsenoside Rd inhibits migration and invasion of tongue cancer cells through H19/miR-675-5p/CDH1 axis

Abstract Objective Tongue squamous cell carcinoma (TSCC) is an oral cancer, with high malignancy and frequent early migration and invasion. Only a few drugs can treat tongue cancer. Ginsenoside Rd is a ginseng extract with anti-cancer effects. Many noncoding RNAs are abnormally expressed in tongue cancer, thus influencing its occurrence and development. H19 and miR-675-5p can promote cancer cell growth. This study aimed to analyze the regulation effect of ginsenoside Rd on H19 and miR-675-5p in tongue cancer. Methodology We used CCK8 and flow cytometry to study the growth and apoptosis. Transwell assay was used to assess invasion; wound-healing assay to assess migration; and colony formation assays to test the ability of cells to form colonies. H19, miR-675-5p, and CDH1 expressions were analyzed by qPCR. E-cadherin expression was detected using western blot. CRISPR/cas9 system was used for CDH1 knockout. Results Ginsenoside Rd inhibited the growth and increased the apoptosis of SCC9 cells. Ginsenoside Rd also inhibited the migration and invasion of SCC9 cells. H19 and miR-675-5p were highly expressed, while CDH1 and E-cadherin expressions were low. H19 and miR-675-5p promoted SCC9 metastasis. In contrast, CDH1 and E-cadherin inhibited the metastasis of SCC9 cells. Bioinformatics analysis showed that miR-675-5p was associated with CDH1. H19 and miR-675-5p expressions decreased after ginsenoside Rd treatment, while CDH1 and E-cadherin expressions increased. Conclusions Ginsenoside Rd inhibits tongue cancer cell migration and invasion via the H19/miR-675-5p/CDH1 axis.

Quality evaluation of different origins and commercial grades of Panax notginseng by HPLC and grey correlation analysis / 中草药

Objective: To establish a method for determination of five saponins in Panax notginseng by HPLC and comprehensively evaluate the quality of it by using grey correlation analysis. Methods: The content of notoginsenoside R1 and ginsenoside Rg1, Re, Rb1, Rd in the different origins and commercial grades of P. notginseng was simultaneously determined by HPLC, and the entire quality evaluation model was established by grey correlation analysis. Results: The established method was applied to quantify five major bioactive components in P. notginseng simultaneously with satisfactory results. Gray correlation method can distinguish the samples from genuine producing areas, qualified samples and unqualified samples, and provide reference for quality evaluation of P. notoginseng and quality evaluation of multi-index components of Chinese materia medica. Conclusion: This HPLC method was simple, accurate, stable and rapid with better separation effect, which was suitable for determination of notoginsenoside R1 and ginsenosides Rg1, Re, Rb1, Rd; The grey recognition analysis was suitable for the comprehensive quality evaluation of multi-component samples of Chinese materia medica.

Ginsenoside Rd improves behavioral impairment of rats with acute plateau status by modulating synaptic plasticity / 中草药·英文版

Objective: To investigate the protective effect of ginsenoside Rd on the improvement of the behavior and synaptic plasticity in rats with acute plateau status. Methods: A total of 60 Wistar rats were randomly divided into the control group, the model group, and the intervention group, with 20 rats in each group. The model was established in low-pressure oxygen chamber simulating the plateau, and the intervention group was administered with ginsenoside. Electron microscope was used to observe synaptic ultrastructure of hippocampal CA1 area, and analyze the structural parameters on the Gray I synaptic interface. Morris water maze and Y electric maze experiment were used for behavioral detection. Results: Compared with the control group, the number of electrical stimulation required for rat to avoid was increased in the model group, the latency in the Morris water maze was prolonged, the swimming distance was increased, and the frequency of crossing the platform was decreased. Under the electron microscope, the synaptic cleft was increased, the length of the synaptic active area was shorter, the postsynaptic density (PSD) was thinner, the flat synapse was increased, and the concave and perforated types were significantly reduced. Compared with the model group, the number of electrical stimulation required for rat to avoid was decreased in the intervention group, the latency in the Morris water maze was shortened, the swimming distance was decreased, and the frequency of crossing the platform was increased. Under the electron microscope, the synaptic cleft was decreased, PSD was thicker, the flat synapse was decreased, and the concave and perforated types were increased. Conclusion: Low pressure and low oxygen environment of plateau damages the plasticity changes of the synaptic structure and function. And to a certain extent, ginsenoside Rd reverses Gray I synaptic interface structure parameters, so as to improve the behavior performance of model rats at high altitude condition.

Determination of seven saponins in flower buds of Panax ginseng by quantitative analysis of multi-components by single marker / 中草药

Objective: On the basis of simultaneous determination of seven saponins in flower buds of Panax ginseng, a method of quantitative analysis of multi-components by single marker (QAMS) for the determination of seven saponins was established, and the feasibility of the method was verified. Methods: Using HPLC-UV, ten batches of dried P. ginseng flowers were used as the research object. Ginsenoside Re was used as internal reference to determine the relative correction factor of ginsenoside Rg1, Rg2, Rb1, Rc, Rb2 and Rd. The content of each component was measured by the traditional external standard method, and the difference between the calculated value and the measured value was compared to verify the feasibility and accuracy of the external standard method. Results: The relative correction factors of six ginsenoside Rg1, Rg2, Rb1, Rc1, Rb2, and Rd in P. ginseng flower were 1.07, 1.05, 0.81, 0.80, 0.64, and 0.84, respectively. The relative correction factors of six ginsenosides were reproducible in the 10 batches, the determiation of QAMS were not significantly different from those measured by the external standard method. Conclusion: In the case of shortage of ginsenoside reference substance, a method of QAMS can be used, the content of ginsenoside Rg1, Rg2, Rb1, Rb1, Rb2, and Rd in flower buds of P. ginseng can be determined by relative calibration factor.

Ginsenoside Rd inhibits IL-1ß-induced inflammation and degradation of intervertebral disc chondrocytes by increasing IL1RAP ubiquitination

Many compounds of ginsenosides show anti-inflammatory properties. However, their anti-inflammatory effects in intervertebral chondrocytes in the presence of inflammatory factors have never been shown. Increased levels of pro-inflammatory cytokines are generally associated with the degradation and death of chondrocytes; therefore, finding an effective and nontoxic substance that attenuates the inflammation is worthwhile. In this study, chondrocytes were isolated from the nucleus pulposus tissues, and the cells were treated with ginsenoside compounds and IL-1β, alone and in combination. Cell viability and death rate were assessed by CCK-8 and flow cytometry methods, respectively. PCR, western blot, and immunoprecipitation assays were performed to determine the mRNA and protein expression, and the interactions between proteins, respectively. Monomeric component of ginsenoside Rd had no toxicity at the tested range of concentrations. Furthermore, Rd suppressed the inflammatory response of chondrocytes to interleukin (IL)-1β by suppressing the increase in IL-1β, tumor necrosis factor (TNF)-α, IL-6, COX-2, and inducible nitric oxide synthase (iNOS) expression, and retarding IL-1β-induced degradation of chondrocytes by improving cell proliferation characteristics and expression of aggrecan and COL2A1. These protective effects of Rd were associated with ubiquitination of IL-1 receptor accessory protein (IL1RAP), blocking the stimulation of IL-1β to NF-κB. Bioinformatics analysis showed that NEDD4, CBL, CBLB, CBLC, and ITCH most likely target IL1RAP. Rd increased intracellular ITCH level and the amount of ITCH attaching to IL1RAP. Thus, IL1RAP ubiquitination promoted by Rd is likely to occur by up-regulation of ITCH. In summary, Rd inhibited IL-1β-induced inflammation and degradation of intervertebral disc chondrocytes by increasing IL1RAP ubiquitination.

Simultaneous determination of six active components in Panax ginseng by QAMS method / 中草药

Objective: This study aimed to simultaneously determine six composition of Panax ginseng by quantitative analysis of multi-components with a single-marker (QAMS) in different paris. Methods: Phenomenex Luna C18 (250 mm × 4.6 mm, 5 μm) was used with mobile phase consisting of acetonitrile-0.1% phosphoric acid for gradient elution at a flow rate of 1.0 mL/min, The column temperature was 25 ℃ and the detection wavelength was 203 nm. Ginsenoside Rb1 was used as reference to establish its relative correction factor of Rg1, Re, Rc, Rb2, Rd, The contents of six components were determined by both external standard method and QAMS. and t test was used to evaluate the feasibility and applicability of QAMS. Results: In a certain linear range, the relative correction factor (RCF) was good, No significant differences were observed between the quantitative results of the two methods. Conclusion: It is feasible and suitable to evaluate the quality of Panax ginseng. It can provide a useful reference to quality control of multi-indexed components in Chinese herbs and traditional Chinese preparations.

Regulatory mechanism of ginsenoside Rd on histone H3 acetylation / 中草药

Objective Based on quantitative proteomics analysis and molecular biology experimental verification, the regulatory mechanism of ginsenoside Rd on histone H3 acetylation levels was elucidated. Methods The effects of ginsenoside Rd on the dynamic changes of proteome of HEK293T cells were detected by.stable isotope labeling with amino acid (SILAC) technique and LC-MS/MS; Quantitative proteomics database analysis was used to monitor the changes in histone acetyltransferase HATs and histone deacetylase HDACs expression levels. Western blotting and qRT-PCR were used to verify the changes of related protein expression and transcriptional level. Gene knockdown experiments were performed using siRNAs to determine the role of ginsenoside Rd in regulating the level of acetyl modifications at histone H3K9 and K18 sites. Results The histone H3K9ac, H3K18ac expression levels in HEK293T cells decreased after ginsenoside Rd treatment, but the P300 catalytic modification of these two sites did not change significantly; At the same time, ginsenoside Rd up-regulated the transcription and expression of HDAC2, and siHDAC2 treatment reversed the down-regulation effects of ginsenoside Rd on H3K9ac and H3K18ac in HEK293T cells. Conclusion Ginsenoside Rd down-regulates the acetylation level of lysine at histone H3K9 and K18 sites by up-regulating HDAC2, thereby affecting transcriptional activation of downstream genes.

Simultaneous determination of 11 components in Naofuqing Capsule by HPLC-MS/MS / 中草药

Objective: To develop a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for simultaneous determination of 11 components including eleutheroside E, 2,3,5,4’-tetera-hydroxystilbene-2-O-β-D-glucoside (stilbene glycoside), isofraxidin, rosmarinic acid, notoginsenoside R1, salvianolic acid B, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rd, and emodin in Naofuqing Capsule. Methods: The analysis was performed on an Agilent Poroshell 120 EC-C18 column (75 mm × 4.6 mm, 2.7 μm), and the mobile phase consisted of water containing 0.1% formic acid and acetonitrile with gradient elution at the flow rate of 0.4 mL/min. The column temperature was maintained at 35 ℃. The MS detection for the 11 tested components was performed in negative ion multiple reaction monitoring mode. Results: Good linear relationship were observed in the test range for 11 components, and the recoveries of eleutheroside E, stilbene glycoside, isofraxidin, rosmarinic acid, notoginsenoside R1, salvianolic acid B, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rd, and emodin were 106.4%, 104.2%, 99.9%, 102.3%, 104.1%, 98.3%, 108.9%, 103.6%, 105.8%, 101.9%, and 104.2%, respectively. The contents of 10 batches of the 11 components were in the ranges of 0.274-0.310, 1.579-1.642, 0.093-0.099, 0.331-0.352, 1.115-1.229, 1.663-1.870, 4.884-5.173, 0.691-0.762, 2.974-3.358, 1.053-1.493, and 0.100-0.115 mg/g, respectively. Conclusion: The established method is sensitive, stable, and rapid. It is suitable for the simultaneous determination of multiple components in Naofuqing Capsule.

Determination of ginsenoside Re, pseudo-ginsenoside F11, ginsenoside Rb3, and ginsenoside Rd from flower buds of Panax quinquefolium / 中草药

Objective To establish the determination method of four ginsenosides from flower buds of Panax quinquefolium by Waters Acquity UPLC H-Class with Xevo TQD. Methods The chromatographic separation was performed on a Waters Acquity UPLC BEH C18 column (50 mm × 2.1mm, 1.7 μm). The mobile phase was a mixture of acetonitrile and water containing 0.05% ammonium hydroxide with gradient elution; The flow rate was 0.45 mL/min, and the column temperature was 35 ℃; Multiple reaction monitoring (MRM) acquisition under negative ion scan mode by ESI was also performed. Results The method was established with good precision, stability, repeatability, and accuracy. Ginsenoside Re, pseudo-ginsenoside F11, ginsenoside Rb3, and ginsenoside Rd showed a good linearity in the range of corresponding concentrations. Conclusion The UPLC-MS/MS method is rapid, simple, accurate, reliable, which can be used for the analysis of ginsenosides from flower buds of P. quinquefolium.

Research Progress in the Effects of Notoginseng Radix et Rhizoma and Its Main Saponins in Cardiovascular Diseases / 中国中医药信息杂志

Blood stasis is the main pathological factor in the occurrence of cardiovascular diseases. Notoginseng Radix et Rhizoma is the representative medicine for activating blood and dissipating stasis. The effective components of panax notoginseng saponins, the active part of Notoginseng Radix et Rhizoma, are ginsenoside Rg1, ginsenoside Rb1, ginsenoside Rd, and notoginsennoside R1, having good intervention effects on cardiovascular diseases, which have become the research hotspots in recent years. This article reviewed the research progress in Notoginseng Radix et Rhizoma and its major saponins in the field of cardiovascular diseases, and provided ideas for clinical prevention and treatment and research and development of medicine.

Establishment of a rapid method to evaluate ultrafiltration membrane aperture for pharmaceutical industry / 中草药

Objective: To solve the technical bottleneck in the application of membrane technology in pharmaceutical industry and to establish a fast and feasible method for evaluating molecular weight cut-off (MWCO) of ultrafiltration membrane. Methods: Based on works, Panax notoginseng saponins (PNS), R1, Rg1, Rb1, and Rd, were selected as indexes for filtering with series ultrafiltration membranes, the retention rate of PNS was accumulated and analyzed with different MWCOs (1×103-1 × 105). Results: The relevant equations were fitted by linear and exponential function, the regression coefficients of equations between the retention rates of PNS and MWCOs were bigger than 0.98. Then, the equations were applied for calculating the different MWCOs of being-measured ultrafiltration membranes. Conclusion: Preliminary judgment is obtained by comparison analysis of cumulative retention rate, and then the optimal MWCO under tested membranes is calculated by subsection equations. This research results have provided a simple and rapid method of evaluating the membrane MWCOs for pharmaceutical industries.

In vivo pharmacokinetic study on total saponins from roots of Panax ginseng in rats / 中草药

Objective: To study the pharmacokinetic profiles of the nine ginsenosides from the roots of Panax ginseng in rats, such as ginsenosides Rb1, Rb2/Rb3, Rc, Rd, Re, Rf, Rg1, and Rh1. Methods: After different time points of ig administration of 200 mg/kg ginsenosides, the blood was taken from the venous plexus of fundus. The biological samples were extracted with n-butanol. Chromatographic separation was performed on a C18 column using a gradient elution program at the flow rate of 0.2 mL/min. The LC-MS system was operated using an electro-spray ionization probe in the negative ion model. After the oral administration of 200 mg/kg ginsenosides to rats, plasma was collected and analyzed under the above conditions. The pharmacokinetic parameters were calculated by non-compartment model. Results: After the oral administration of ginsenosides to rats, six ginsenosides were detected in plasma which included Rb1, Rb2/Rb3, Rc, Rd, Re, and Rg1. Among these ginsenosides, the protopanaxatriol ginsenoside Rg1 and Re were quickly eliminated. However, the pharmacokinetic behaviors of protopanaxadiol ginsenoside Rb1, Rb2/Rb3, Rc, and Rd were markedly different from those of ginsenosides Rg1 and Re in rats with the significantly longer half-life of the protopanaxadiol ginsenosides. Conclusion: The method is accurate, stable, and reliable, and can be used for profiling total ginsenosides' pharmacokinetic properties in rats.

Effects of different steaming methods on saponins in taproots of Panax notoginseng / 中草药

Objective: To research the effect of different steaming methods on the saponins content in the taproots of Panax notoginseng. Methods: Colorimetric method is used to determine the content of total saponins in the taproot of P. notoginseng, using high performance liquid chromatography (HPLC) method for determining the content of monomer saponins in the taproot of P. notoginseng. Results: The contents of total saponins, notoginsenoside R1, and ginsenosides Rg1, Re, Rb1, and Rd in the taproot of P. notoginseng have been reduced to some extent, while the contents of five kinds of monomer saponins of ginseng, i.e. ginsenosides Rh1, Rh4, Rk3, 20(S)-Rg3, and 20(R)-Rg3 all have been increased in varying degrees after steamed. Conclusion: The different steaming methods have the different influences to saponin composition in the taproot of P. notoginseng, the contents of total saponins and five main saponins could decrease and new generated monomer saponins could increase associated with the steaming time and temperature. This method can be used for the determination of the contents of saponin compounds and quality control in processed notoginseng products, and provide the basis for the study on the correlation between notoginseng "sheng da shu bu" and substance changes, while provide the certain theory basis for researching the accumulation rule of rare saponins.

Determination of eight components in Qibai Pingfei Granule by quantitative analysis of multi-components with a single-marker / 中草药

Objective: To establish a quantitative analysis of multi-components with a single-marker (QAMS) method for the determination of eight components in Qibai Pingfei Granule (QPG). Methods: Ginsenoside Rb1 was used as the internal reference substance. The relative correlation factors (f) of ginsenosides Rg, Re, Rf, Rc, Rb2, Rc, and astragaloside were calculated and established. The results were compared with those obtained by the external standard method in 10 batches of QPG. The results showed that the f values of ginsenosides Rg, Rf, Rc, Rb2, astragaloside, and ginsenosides Rc to ginsenoside Rb1 were 1.244, 1.075, 1.133, 1.090, 1.071, 0.967 and 1.070. Results: Ginsenosides Rg1, Re, Rf, Rb1, Rc, Rb2, Rd, and astragaloside had good relations within the ranges of 0.416 0-4.992, 0.315 4-3.785 2, 0.259 6-3.115, 0.385 2-4.622, 0.222 8-2.674, 0.193 2-2.318, 0.183 5-2.202, and 0.574 6-6.895 μg, respectively. The two methods did not show the significant difference in assay results. Conclusion: So the QAMS method is feasible and credible, and could be used to determine the multiple components in QPG.

Simultaneous determination of sixteen ginsenosides in Panax ginseng and its preparation by HPLC / 中草药

Objective: In order to evaluate the quality of Panax ginseng and its preparation, a simple and accurate HPLC method for determining the contents of 16 ginsenosides from P. ginseng was established. Methods: The chromatographic separation was achieved on a C18 column (150 mm × 4.6 mm, 5 μm) using a mobile phase made up of acetonitrie and water at a flow rate of 1.0 mL/min. The detection wavelength and column temperature were set as 203 nm and 35°C, respectively. Results: Sixteen ginsenosides (Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, Rb3, F1, Rd, F2, Rg3, protopanaxatriol, compound K, Rh2, and protopanaxadiol) were separated at baseline with good linearity (r ≥ 0.9990). The recovery rates were 95%-102% (RSD < 2%). Conclusion: The method is simple, fast, accurate, and could be applied to the quality control of P. ginseng and its preparation.

The growth and biotransformation characteristics of a high ginsenoside Rd productive mutant strain Paecilomyces bainier sp229-7 / 复旦学报（医学版）

Objective To investigate the mycelia growth of the mutant strain Paecilomyces bainier sp229-7 and it's biotransformation from ginsenoside Rb1 to Rd. Methods The substrate was biotransformed by the mutant in different conditions, and the products were analyzed by HPLC. Results The period of vigorous growth of the mutant was 12-48 h. The optimal transformation temperature range was from 27 ℃ to 32 ℃. The best time for adding substrate was 36 h after inoculation and the substrate (Rb1 20 mg/mL) was specifically transformed into ginsenoside Rd. The transformation process was terminated at 72 h. The ginsenoside Rb1 could be transformed to Rd by all of mycelia, crude enzymes and external enzymes. Conclusions Compared with the reference, the mutant of Paecilomyces bainier sp229-7 growth is quicker and the transformation efficiency has been greatly improved. The reaction time can be shortened to 3 days, and the amount of substrate increased at least 20 times.

Determination of ginsenoside Rb_1,Re,Rb_2 and Rd in Shenqi Granules by microbore liquid chromatography / 中成药

AIM: To develop a method for determining ginsenoside Rb_1,Rc,Rb_2 and Rd in Shenqi Granules (Radix Rhizoma Ginseng,Radix Astragali,Rhizoma Atractylodis macrocephalae,Poria,etc.) by microbore LC.METHODS: Microbore liquid chromatography was applied.Microsil C_(18) column( 150 mm×1.0 mm) was used with the mobile phase containing acetonitrile-water for gradient elution.The flow rate was 50 μL/min,and the detection wavelength was set at 220 nm.RESULTS : The relationships between the concentrations and the peak areas of these for components were all linear.The recoveries were 98.14% for ginsenoside Rb_1,98.04% for ginsenoside Rc,98.79% for ginsenoside Rb_2,and 97.97% for ginsenoside Rd respectively.The relative standard deviations (RSD) were 0.67%,0.99%,0.75%,and 1.46%,respectively.CONCLUSION: This method is simple,convenient and can be used for quality control.

Effect of Ginsenoside Rd Pretreatment on the Expressions of NR2B Receptor and Endonuclease G After Focal Cerebral Ischemia-Reperfusion in Rats / 国际脑血管病杂志

Objective:To observe the effect of ginsenoside Rd pretreatment on the expressions of N-methyl-D-aspartate(NMDA)receptor subunit NR2 B protein and endonuclease G(EndoG)in basal ganglia region after cerebral focal ischemia-reperfusion in rats and to investigate possible mechanism of ginsenoside Rd in the treatment of ischemic stroke.Methods:A rat model of middle cerebral artery occlusion(MCAO)was induced by intraluminal filament method.The expressions of NR2B and EndoG in basal ganglia region for focal cerebral iSChemia 1 hour,and 1,6,24 and 72 hours reperfusion were detected by immunohistochemical staining and image analysis method.The effects of ginsenoside Rd on the expressions of FaxioG and NR2B and the volume of cerebral infarction were evaluated.Results:The positive expression of NR2B in basal ganglia region on the ischemic side in ischemia-reperfusion group was increased significantly.The expression of EndoG in the nucleus was notable;the positive expressions of NR2B and EndoG at different reperfusion time points in ginsenoside Rd pretreatment group were decreased significantly(P<0.05 or P<0.01),and the volume of cerebral infarction was reduced significantly(P<0.01).Conclusions:The expressions of NMDA receptor subunit NR2B protein and apoptosis-inducing factor EndoG were increased significantly after cerebral focal ischemia reperfusion;ginsenoside Rd pretreatment may significantly reduce the expressions of NR2B and EndoG.It reduces the volume of cerebral infarction by inhibiting excitatory neurotoxicity and blocking neuronal apoptosis,and thus plays a role in neuroprotection.