Extraction of Astragalus Polysaccharides and Ganoderma lucidum Mycelia Polysaccharides and Their in Vitro Antioxidative Effects

[Objectives]To explore the extraction and in vitro antioxidant effects of astragalus polysaccharides(APS)and Ganoderma lucidum mycelia polysaccharides(GLMPS).[Methods]By studying the polysaccharides of the herbal medicinal material Astragalus membranaceus and the fungal medicinal material Ganoderma lucidum mycelia,two polysaccharides were mixed according to different proportions and concentrations by using the principle of traditional Chinese medicine compound combination.The effect of polysaccharides on the scavenging ability of hydroxyl radical system was determined by salicylic acid method.[Results]When the compound ratios of GLMPS and APS were 1∶1,1∶4,1∶5,4∶1,and 5∶1,the scavenging effect of compound polysaccharides was better than that of single-component polysaccharides,and with the increase of concentration,the scavenging effect increased.When the ratio of GLMPS and APS was 5∶1,the hydroxyl radical scavenging rate of the compound polysaccharide reached 59.77%,which was 18.72%higher than that of single GLMPS and 28.58%higher than that of single APS.The scavenging effect of compound polysaccharide is closely related to the compound ratio and concentration.[Conclusions]APS and GLMPS can obtain better hydroxyl radical scavenging ability than single-component polysaccharides through compounding in appropriate proportions.In addition,within a suitable concentration range,as the concentration increases,the scavenging ability also increases.

Studies on Active Polysaccharides from Ganoderma lucidum vith Immune Activity

从云头状灵芝[Ganoderma Lucidum(Leyss.ex Fr.)Karst.]中分离得到三种灵芝多糖BN3A，BN3B及BN3C，它们均表现明显免疫调节作用。从灵芝多糖BN3B及BN3C中各分离得到四个多糖均一体，对其中主要成分BN3C1，BN3C3，BN3B1及BN3B3进行了物理及化学研究，它们的平均分子量依次为1.6×10^4，2.5×10^4，3.5×10^4及4.0×10^4。经完全酸水解、红外光谱测定、过碘酸氧化、甲酸生成、Smith降解等证明，BN3B1及BN3C1均为β－（1→6)(1→3）甙键相连的葡聚糖。BN3B3为阿拉伯半乳聚糖。BN3C3为由葡萄糖和阿拉伯糖组成的肽多糖。它们均为β－（1→6）（1→3）甙键相连。

Related gene expressions in anti-keratinocyte aging induced by Ganoderma lucidum polysaccharides

Objective:To examine the level of expression of anti skin aging gene of Ganoderma lucidum polysacchayides and clarify its mechanism with anti aging of this ancient Chinese medicine.Methods:HacaT cell of keratinocytes lines were cultured and treated with the polysaccharides.The total RNA was extracted with Trizol reagent and cDNA was synthesized by reverse thanscription.The obtained cDNAs were then fluorescently labeled with cy3 and cy5 respectively and hybridized with gene expressing pedigree cDNA chip.The images were scanned and analyzed with special software.The scan data were analyzed with software and checked by real time PCR.Results:Among total 18 346 human genes,the expression of 103 ones was up-regulated and 378 ones down-regulated.It was demonstrated evidently that Ganoderma lucidum polysaccharides affected the expression of genes of anti skin aging.Two ways are anastomotic.Conclusion:it is concluded by analysis of function of these up-regulation and down-regulation genes that Ganoderma lucidum polysaccharides may play an important role in boosting cell growth and against skin aging.It shows that the results of gene array reliable by real time PCR.

GPP(composition of Ganoderma lucidum polysaccharides and Polyporus umbellatus polysaccharides) protects against DSS-induced murine colitis by enhancing immune function and regulating intestinal flora

Previous study have demonstrated that a compound composed of water-soluable Ganoderma lucidum polysaccharides(GLP)and Polyporus umbellatus polysaccharides(PUP)in a ratio of 3:1 named GPP enhances innate immune function in mice through enhancing the function of macrophage cells and activity of natural killer(NK)cells.Here in our research,we further investigated the effect of GPP on the diversity and composition of intestinal flora,and explored its effect on colitis model mice.The immunoregulatory verification experiments of GPP were conducted in both normal and DSS-induced mice model.Our research showed that GPP increased the diversity of intestinal microorganisms in mice with the extension of administration time.Daily GPP intake attenuated DSS-induced colon injury,protected the splenic lymphocyte proliferation ability,enhanced the serum hemolysin synthesis,and increased peripheral phagocytes and NK cell activity in model mice.Comparisons of the predominant gene pathways of the bacterial microbiota showed that DNA repair and recombination,base mismatch repair pathways was stronger in GPP-treatment group than in control group,indicating the possible molecular mechanisms of immune function regulation.Our study showed that GPP regulated immune function in both health and colitis model,and had a positive effect on maintaining intestinal flora homeostasis.

The Impact on Extraction Effect of Polysaccharide from Ganoderma lucidum by Different Pretreatment

[Objectives] The research aimed to study the impact on extraction effect of polysaccharide from Ganoderma lucidum( lingzhi) by different pretreatment methods. [Methods] The impacts on extraction of G. lucidum polysaccharide by soaking,microwave and air flow fine pulverization were contrasted,and the extraction effect of G. lucidum polysaccharide by combining the optimal pretreatment manner with hot water extraction,alcohol extraction,alkali extraction,ultrasonic binding enzyme extraction,and microwave extraction was compared. Finally,the property of G. lucidum polysaccharide obtained after air flow fine pulverization pretreatment was detected and analyzed by high performance liquid chromatography. [Results] The optimal pretreatment method was air flow fine pulverization. Compared with traditional method-direct extraction( coarse grinding combining hot water extraction),crude yield changed little,while polysaccharide content and yield were improved by 114% and 104%. The best combination manner was air flow fine pulverization pretreatment combining with alkali extraction. Compared with traditional method,crude yield,polysaccharide content and yield were improved by 76%,78% and 215% respectively. The property of G. lucidum polysaccharide obtained after air flow fine pulverization pretreatment was detected and analyzed by high performance liquid chromatography. It was found that the treatment method had little impact on the property of G. lucidum polysaccharide. [Conclusions]Air flow fine pulverization pretreatment could greatly improve extraction effect of G. lucidum polysaccharide,which mainly improved the content and yield of G. lucidum polysaccharide,and extraction was more complete,with less impact on the property of the extracted polysaccharide. It was speculated that air flow fine pulverization pretreatment mainly destroyed mechanical support wall membrane structure of G. lucidum fine powder,making that intracellular functional substances completely dissolved out of the cell,and the content would be studied in follow-up experiment.

Ganoderma lucidum polysaccharide peptide alleviates hepatoteatosis via modulating bile acid metabolism dependent on FXR-SHP/FGF

OBJECTIVE Ganoderma lucidum polysaccharide peptide(GLPP)is a group of extract from Ganoderma lucidum with a molecular mass of approximately 5×10^5,which ratio of polysaccharide to peptide is approximately 95%/5%.The purpose of this study was to determine whether GLPP has therapeutic effect on Non-alcoholic fatty liver disease(NAFLD).METHODS Ob/ob mouse model and ApoC3 transgenic mouse model were used for exploring the effect of GLPP on NAFLD.Key metabolic pathways and enzymes were identified by metabolomics combining with KEGG and PIUmet analyses and key enzymes were detected by Western blotting.Hepatosteatosis models of HepG2 cells and primary hepatocytes were used to further confirm the therapeutic effect of GLPP on NAFLD.RESULTS GLPP administrated for a month alleviated hepatosteatosis,dyslipidemia,liver dysfunction and liver insulin resistance.Pathways of glycerophos⁃pholipid metabolism,fatty acid metabolism and primary bile acid biosynthesis were involved in the therapeutic effect of GLPP on NAFLD.Detection of key enzymes revealed that GLPP reversed low expression of CYP7A1,CYP8B1,FXR,SHP and high expression of FGFR4 in ob/ob mice and ApoC3 mice.Besides,GLPP inhibited fatty acid synthesis by reducing the expression of SREBP1c,FAS and ACC via a FXR-SHP dependent mechanism.Additionally,GLPP reduced the accumulation of lipid droplets and the content of TG in HepG2 cells and primary hepatocytes induced by oleic acid and palmitic acid.CONCLUSION GLPP significantly improves NAFLD via regulating bile acid synthesis dependent on FXR-SHP/FGF pathway,which finally inhibits fatty acid synthesis,indicating that GLPP might be developed as a ther⁃apeutic drug for NAFLD.

Effects of Ganoderma lucidum polysaccharides peptide on tumor metastasis in mice with sleep fragmentation

In recent years,studies have shown that there is a correlation between sleep disorders and cancer,but at the present stage,the research on sleep disorders and tumor related animal models is relatively insufficient.Our research will focus on mice bearing B16F10-luc-G5 melanoma tumor with sleep fragmentation,detecting promoting effect of sleep fragmentation(SF)on the metastasis of melanoma.At the same time,we used Ganoderma lucidum poly⁃saccharides peptide(GL-pp,80 mg·kg-1),a component of traditional Chinese medicine Ganoderma lucidum,which has long enjoyed a good reputation at home and abroad,to observe its anti-tumor metastasis effects on B16F10-luc-G5 mice with SF.Then we used whole proteomics to analyze the difference proteins expressed in lung tissue and compared between groups,includes mice bearing B16F10-luc-G5,mice bearing B16F10-luc-G5 with SF and GL-pp administered mice bearing B16F10-luc-G5 with SF.With the analysis using bioinformatics,we found several key proteins,their genes name are Adcy9,ptk2,Yap1 and Lpin2,Per1 and Tim.And several important clusters,they are,immune system,platelet aggres⁃sion,energy metabolism,cell cytoskeleton,cell adhesion and circadian rhythms.Moreover,we detected the TLR4 signal pathway and macrophage differentiation to reconfirm the results of proteomics and trying to elucidate the mechanism of SF on tumor growth and metastasis and the effects of GL-pp.

Effect of Ganoderma lucidum Polysaccharide on Antioxidative Ability of Rats with Myocardial Injury Induced by Doxorubicin

Objective: The objective is to investigate the effect of Ganoderma lucidum polysaccharide (GLPS) on the oxidative stress induced by doxorubicin (DOX) in cardiomyocytes. Methods: SD rats were divided into control group, DOX group, GLPS low dose and high dose + DOX group. SOD and MDA levels in myocardial tissue were detected in each group. The expression of related proteins in each group was detected by Western blot. Results: GLPS can increase the SOD level and decrease MDA caused by DOX (p Conclusion: Ganoderma polysaccharide can improve the oxidative stress injury of myocardial tissue induced by doxorubicin and play a protective role in myocardial tissue.

Hydrodynamic Shear Stress Affects Cell Growth and Metabolite Production by Medicinal Mushroom Ganoderma lucidum

By investigating the shear effect on submerged cultivation of a traditional Chinese medicinal herb Ganoderma lucidum, a relatively high cell concentration of 13.8 g·L-1 by dry mass was obtained in bioreactor at an impeller tip speed (ITS) of 0.51m·s-1. At an ITS of 0.51,1.02 and 1.53m·s-1, a maximal production titer of intracellular polysaccharide was 2.64, 2.20 and 2.28g·L-1 and that of ganoderic acid was 306, 299 and 273g·L-1, respectively. Under these ITSs, the maximal mean projected area of dispersed hyphae was 3.70, 2.54 and 2.13×104μm2, and that of pellets was 0.91, 0.67 and 0.55mm2, respectively. The information obtained is useful for efficient submerged cultivation of mushrooms on a large scale.

Ganoderma lucidum polysaccharide inhibits LPS-induced inflammatory injury to mammary epithelial cells

This study sought to investigate whether Ganoderma lucidum polysaccharide(GLP)has a protective effect on lipopolysaccharide(LPS)-induced inflammatory injury to mammary epithelial HC-11 cells and to characterize the mechanism involved.Cell viability was assessed using the cell counting kit 8(CCK-8)method,tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and IL-1βlevels were measured by enzyme linked immunosorbent assay(ELISA),and IκBa,p65 NF-κB and STAT3 mRNA were determined using quantitative reverse transcription PCR(qRT-PCR),p65 and STAT3 protein expression were determined using Western blotting,respectively.GLP was shown to inhibit LPS-induced TNF-α,IL-6,and IL-1βproduction(P<0.01 or P<0.05),GLP was also shown to increase IκBαmRNA expression(P<0.01),decrease p65 and STAT3 mRNA expression(P<0.01 or P<0.05),and decrease p-p65,p65,p-STAT3,and STAT3 protein expression in breast epithelial cells(P<0.01 or P<0.05).The findings suggest that GLP inhibits nuclear factor kappa-B(NF-κB)and signal transducers and activators of transcription(STAT)signaling by preventing IκBαdegradation and p65 and STAT3 phosphorylation.This results in lower LPS-induced TNF-α,IL-6,and IL-1βproduction and prevents inflammatory cell injury.

Ganoderma atrum polysaccharide relieves mitochondrial dysfunction to alleviate hydrogen peroxide-induced senescence via activating autophagy

Polysaccharides from genus Ganoderma have prominent anti-aging effect,but their mechanisms are incompletely clarified.In our previous experiments,Ganoderma atrum polysaccharide(PSG)was exhibited to significantly alleviate senescence and DNA damage of A375 cells.To investigate its underlying mechanism,we conducted RNA-Seq analysis and found that PSG upregulated autophagy and mitochondria pathways compared to the model group.Further experiments showed that PSG relieved mitochondrial dysfunction via inhibiting the accumulation of reactive oxygen species(ROS)and loss of the mitochondrial membrane potential(MMP).Meanwhile,PSG activated autophagy and enhanced lysosomal function through transcription factor EB(TFEB).Interestingly,the pretreatment of autophagy inhibitors bafilomycin A1(Baf A1)reversed the ROS decline induced by PSG,which subsequently increased cellular senescence of A375.In addition,PSG had a similar anti-aging effect on normal fibroblast WI-38.Taken together,PSG might reduce ROS levels through activating autophagy,which in turn suppressed mitochondrial dysfunction and cellular senescence.

Effects of GLP on Intestinal Mucosal Injury and the Change of TNF-α Content in Hemorrhagic Shock Rabbits

Objective To observe the intestinal mucosal injury and the change of TNF-αcontent in rabbits with hemorrhagic shock/reperfusion(HS-R)and the effects of ganoderma Lcidum polysaccharide(GLP)on them.Methods 30rabbits were made into hemorrhagic shock,then reperfused with different liquids.These rabbits were divided by random number table into three groups:sham operation group(Sham group),reperfusion with normal saline group(NS group),reperfusion with 1%GLP group(LS group).Bacterial translocation of blood and TNF-αcontent in serum were respectively observed at the time before shock,40 min after shock,40 min and 90 min after.TNF-αcontent in intestinal mucosa and the degree of intestinal mucosal injury were examined at 90 min post-resuscitation.Results 1 With the extension of reperfusion time,the positive rate of blood bacteria increased gradually in NS group,which was significantly higher than that of Sham group and LS group(P<0.05),meanwhile the degree of intestinal mucosal injury in NS group was more severe than that of Sham group and LS group too(P<0.05).2TNF-αcontent in serum of NS group and LS group were increased obviously compared with that before shock and in Sham group(P<0.05).TNF-αcontent in serum was further increased after reperfusion with NS,which was distinctly higher than that in LS group.TNF-αcontent in intestinal mucosa of NS group was significantly higher than that in LS group and Sham group too(P<0.05).Conclusion GLP can protect intestinal mucosa against HS-R injury,and its effects may relate with the change of TNF-αin hemorrhagic shock rabbits.