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OBJECTIVE： To study the imp rovement effects of Ganoderma lucidum polysaccharides crude extract on estradiol-induced thymus atrophy in mice. METHODS ：Totally 60 female ICR mice were randomly divided into normal control group（normal saline ），model group （normal saline ），G. lucidum polysaccharides crude extract high-dose and low-dose groups （400，100 mg/kg，by crude drug ），with 15 mice in each group. Except for normal control group ，other groups were given estradiol intraperitoneally （0.1 mg/mice，6 times）every other day to establish thymic atrophy model. The next day after modeling finished，they were given relevant medicine intragastrically ，once a day ，for consecutive 14 d. Twenty-four hours after last medication，organ（thymus，spleen）index，MDA content and GST activity in plasma were determined. HE staining was adopted to observe the pathological changes of thymus and spleen tissue in mice. The thymus cell apoptosis was examined by TUNEL assay ， and the T cell subsets in peripheral blood were detected by flow cytometry. RESULTS ：Compared with normal control group ，the thymus index ，proportion of CD 3+CD4+T cell in peripheral blood and CD 4+/CD8+ ratio were decreased significantly in model group （P＜0.01）；spleen index ，MDA content in plasma and thymocyte apoptosis level as well as the proportion of CD 3+CD8+T cell in peripheral blood were all increased significantly （P＜0.05 or P＜0.01）. Thymic cortex and medullary boundary of mice was blurred；the intercellular space was enlarged ；some cells were damaged and apoptotic in cortex ；no pathological changes were found in the spleen. Compared with model group ，thymus index and GST activity in plasma as well as proportion of CD 3+CD4+T cell in peripheral blood and CD 4+/CD8+ ratio were all increased significantly in G. lucidum polysaccharides crude extract high-dose group（P＜0.05 or P＜0.01）；while MDA content in plasma ，the apoptosis level of thymocytes were all decreased significantly （P＜0.01 or P＜0.05）；and the pathological changes of thymus were improved significantly. MDA content in plasma was decreased significantly in G. lucidum polysaccharides crude extract low-dose group （P＜0.01），and other indexes/pathological changes were not obvious. CONCLUSIONS ：High dose （400 mg/kg）of G. lucidum polysaccharides crude extract can improve the thymus atrophy induced by estradiol in mice.
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Ganoderma lucidum is a traditional folk common rare medicinal herb, which plays an important role in maintaining human health. Among them, G. lucidum polysaccharide is one of the main bioactive components of G. lucidum, which has various pharmacological effects such as anti-cancer, immunomodulatory, antibacterial, antioxidant, and scavenging free radicals. Due to the complex etiology and pathogenesis of central nervous system diseases, the clinical manifestations are diverse, and the individual response to treatment varies widely, which brings great challenges to clinical treatment. The characteristics and mechanism of the central neuroprotective effect of G. lucidum polysaccharide are reviewed in this paper from the aspects of Alzheimer's disease, anti-epileptic, modulating microglia inflammatory response, etc., in order to provide relevant evidence for its clinical application.
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OBJECTIVE Non-alcoholic fatty liver disease(NAFLD) encompasses a series of patho-logic changes ranging from steatosis to steatohepatitis,which may progress to cirrhosis and hepatocel-lular carcinoma.The purpose of this study was to determine whether Ganoderma lucidum polysaccha-ride peptide (GLPP) has therapeutic effect on NAFLD. METHODS ob/ob mouse model and ApoC3 transgenic mouse model were used for exploring the effect of GLPP on NAFLD. Key metabolic path-ways 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 glycerophospholipid 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 hepato-cytes 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 therapeutic drug for NAFLD.
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Objective To extract, separate, and purify polysaccharide from Ganoderma lucidum with alkali from the residue after water extraction, characterize the basic physicochemical properties and structural features in detail, and study the immunomodulatory activity in vitro. Methods The polysaccharide LZJ-015 was isolated and purified from the dry fruiting bodies of G. lucidum by alkaline extraction and ethanol precipitation following Q-Sepharose Fast Flow ion exchange chromatography column. Monosaccharide composition and molecular weight of LZJ-0.15 was analyzed by high performance liquid chromatography (HPLC) with PMP precolumn derivatization and high performance gel permeation chromatography-multiple angle laser (HPGPC-MALLS), respectively. The detailed structure of polysaccharide LZJ-0.15 was characterized by 1H-NMR, 13C-NMR, 1H-1H COSY, and 1H-13C HSQC spectrum. The immunomodulatory activity of G. lucidum polysaccharide was test by RAW264.7 cells phagocytose neutral red experiment. Results The molecular weight, molecular radius, and Mw/Mn of LZJ-0.15 were determined to be 24 700, 46.6 nm, and 1.019, respectively. The monosaccharide composition was confirmed to be mainly composed of glucose (92.3%). LZJ-0.15 was →3) Glc (β1→and→6) Glc (β1→linked glucan indicated by NMR spectrum. Moreover, G. lucidum polysaccharide exhibited good immunomodulatory activity in our study. Conclusion G. lucidum polysaccharide LZJ-0.15 from alkaline extraction showed better immune activity than the polysaccharide extracted from water, which would be potentially developed as an effective immunomodulatory agent.
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Objective To study the effect of ganoderma lucidum polysaccharides on T cell subsets and AQP1, AQP3 expression of bladder cancer T24 cell line bearing mice.Methods 60 BALB/C nude mice were selection as experimental animals, bladder tumor bearing animal models were made by bladder cancer T24 cells subcutaneous injection, and were randomly divided into control group, cisplatin group, ganoderma lucidum polysaccharides and cisplatin group, cisplatin group given 25 mg/kgcisplatin intraperitoneal injection, cisplatin combined ganoderma lucidum polysaccharide group given 200 mg/kg ganoderma lucidum polysaccharide lavage, 25 mg/kg cisplatin intraperitoneal injection, the control group given quite a volume normal saline lavage.Then tumor volume and weight, peripheral blood T cell subsets and AQP1, AQP3, Caspase-3, Bax mRNA content in tumor tissue were determined.Results Cisplatin group, cisplatin combined ganoderma lucidum polysaccharide tumor volume and mass were significantly lower than the control group (P<0.05), cisplatin and ganoderma lucidum polysaccharide tumor volume and mass were significantly lower than that cisplatin group ( P<0.05 ); Cisplatin group, cisplatin combined ganoderma lucidum polysaccharides group CD4 + T cells, CD8 +T cells , CD4 +/CD8 + were significantly higher than that of control group (P<0.05), cisplatin combined ganoderma lucidum polysaccharide group CD4 +T cells, CD4 +/CD8 +were significantly higher than that of cisplatin group (P<0.05); Cisplatin group, cisplatin combined ganoderma lucidum polysaccharide group mices tumor tissues Bax, Caspase 3 mRNA content were significantly lower than the control group (P<0.05), cisplatin combined ganoderma lucidum polysaccharide group Bax,caspase 3 mRNA content were significantly lower than that of cisplatin group (P<0.05); Cisplatin group, cisplatin combined ganoderma lucidum polysaccharide group mice tumor tissue AQP1, AQP3 mRNA content were significantly lower than the control group (P<0.05); Cisplatin combined ganoderma lucidum polysaccharide group AQP1, AQP3 mRNA content were significantly lower than cisplatin group (P<0.05 ). Conclusion ganoderma lucidum polysaccharide can inhibit tumor growth and enhance cellular immune function of bladder cancer T24 cell line bearing mice, and adjust the expression of pro-apoptotic genes, AQP1 and AQP3.
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Objective To observe the effects of ganoderma lucidum polysaccharide(GLP) on intestine mucosal barrier function and tumor necrosis factor-α(TNF-α)in hemorrhagic shock rabbits. Methods Thirty healthy Japanese white rabbits were divided into three groups by random number table:sham operation group, resuscitation/reperfusion with normal saline(NS)group(NS group)and resuscitation/reperfusion with GLP group (GLP group). The hemorrhagic shock models were reproduced by being bled from the femoral artery to the mean arterial pressure(MAP)of 30-40 mmHg(1 mmHg=0.133 kPa)within 10 minutes and kept maintaining at this MAP level for 40 minutes. Then,the anti-shock therapy was given. The rabbits in NS group were immediately and quickly resuscitated with reperfusion of their shed blood and intravenous drip of NS in a volume equal to 2-fold of the volume of the shed blood. The rabbits in GLP group were dealt with the same means as NS group except NS was replaced by 1%GLP. The rabbits in sham operation group were only cannulated and maintained in a status of blood pressure measurement,but without bloodletting and resuscitation. The TNF-α content,the number of positive bacterial culture of blood bacteria were observed at the time before shock(S0),40 minutes after shock(S40)and 40 minutes and 90 minutes after resuscitation/reperfusion(R40,R90),respectively in the three groups. The serum endotoxin level and the TNF-αcontent in intestinal mucosa were examined at R90. Under the light microscopy,the pathological changes of intestinal mucosa were examined,and the score of the mucosal damage was evaluated by the criteria of Chiu method. Results ①Before shock,the results of blood bacterial culture of all animals in three groups were negative. With the extension of time,the positive rates of blood bacteria were increased gradually at R40 and R90 in NS and GLP groups,showing increment of bacterial translocation,but the levels at R40 and R90 were significantly lower in GLP group than those in NS group at the same time points(number of rabbits at the time of R40:2 vs. 4,at the time of R90:4 vs. 6,both P<0.05). Escherichia coli was the most common positive bacteria in the blood;the other positive bacteria were sequentially less and less,Enterococcus,Lactobacillus acidophilus and Staphylococcus aureus. Compared with sham operation group,the serum endotoxin levels of NS and GLP groups were significantly higher at R90(kU/L:1.823±0.963,1.361±0.529 vs. 0.064±0.036,both P<0.05). ② Under the light microscope,it was shown that in NS group the mucosal villi were edematous,the sub-epithelial space was widened,and there was infiltration of neutrophils and lymphocytes. The damage of intestinal mucosa in GLP group was obviously milder than that in the NS group. The Chiu scores of GLP and NS groups were significantly higher than that of sham operation group;meanwhile,Chiu score of GLP group was lower than that of NS group(1.44±0.64 vs. 3.79±1.23,P<0.05).③Along with extension of ischemia and reperfusion time,TNF-α contents in NS and GLP groups were gradually increased significantly,the peak value being at R90,meanwhile,the levels in both groups were significantly higher than the level of sham operation group,and the TNF-α content in plasma of GLP group was lower than that of NS group〔A value:33.350±7.950 vs. 85.080±4.330,P<0.05〕. Compared with sham operation group,the TNF-αcontents in intestinal mucosa of NS and GLP groups were higher than the level in sham operation group,meanwhile,TNF-αcontent in intestinal mucosa of GLP group was lower than that of NS group〔A value:96.38±8.59 vs. 167.73±12.32, P<0.05〕. Conclusion In the salvage process of resuscitation-reperfusion,GLP can protect intestinal barrier function in rabbits with hemorrhagic shock,and its effect may be related to antagonizing the release of TNF-α.
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Objective To study the effects of Ganoderma lucidum polysaccharide (GLP) on PBMCs and the related immune mechanism.Methods PBMCs from cancer patients and healthy donors were isolated and treated with different doses of Ganoderma lucidum polysaccharides (10 ng/ml,50 ng/ml and 100 ng/ml).DC cell costimulatory molecules (HLA-DR,CD83,and CD11 c),Th1 (CD3 + CD8-IFN-γ+) cells,Th2 (CD3 + CD8-IL-4+) cells and NK (CD3-CD56+) were analyzed by FCM.Furthermore,The CD3+ CD4+ Th ceils were separated by immunomagnetic beads and stimulated with Ganoderma lucidum polysaccharides at different concentrations in culture.After 24 h,the cytokine expression levels of Th1 and Th2were detected by RT-PCR.The expressions of Th1 differentiation-related transcription factor,STAT4,were analyzed by Western blot.Results Ganoderma lucidum polysaccharides can significantly stimulate in vitro Thl cell differentiation (P＜0.01) in a dose depend manner.It correlates with an increased expression of STAT4 and the elevated mRNA expression levels of Th1 cytokineincluding IL-12,IFN-γ and TNF-α (P＜0.01).Conclusion Ganoderma lucidum polysaccharides may promote Th1 differentiation and increase the secretion of Th1 cvtokines through the upregulation of STAT4.
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Objective To study the effects on inhibit tumor growth and enhance immune function of Ganoderma lucidum polysaccharide(GLP)combined with low-dose cisplatin in tumor-bearing mice.Methods 2 × 106 Lewis cells in 0.1 mL PBS were injected subcutaneously into the flanks of healthy female BALB/c nude mice (5～6 weeks old).When the tumor volume reached 100 mm3,mice were randomly assigned to 4 groups,control group (Oral gavage of 0.5 ml PBS and intraperitoneal injection of 0.5 ml),GLP group (gavage once every two days,40 mg/kg GLP in 0.5 ml),cisplatin group (intraperitoneal injection once every two days,2 mg/kg cisplatin in 0.5 ml) and the combination group (such as the above dose of cisplatin and GLP).After the treatment regimen was complete,the mice were sacrificed,and the tumors were removed,photographed,weighed,and prepared for histological analysis.The immune cells in spleen and peripheral blood were analyzed by flow cytometry.Results GLP can effectively increase anti-tumor effect of the small dose cisplatin,improve the survival of tumor-bearing mice,promote Th cells convert to Th1 in peripheral blood,increased the expression of CD 11c + on DC cell surface (GLP group 4.59％ VS the control group 1.06％,the combined group 7.21％ VS the cisplatin group 2.8％).Conclusion GLP can improve the immune function of tumor-bearing mice,had synergistic anti-tumor growth with low dose cisplatin,moreover,reduce renal toxic effects of cisplatin.
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Polysaccharide was extracted by boiling water reflux method from the fruiting body of Ganoderma lucidum. Additionally, the purified polysaccharide was obtained by removing protein with Sevage way, ethanol precipitation, centrifugation, run water dialysis, membrane separation, concentration and frozen-drying. The structural characteristics, chain conformation and triple-helix conformation of ganoderma lucidum polysaccharide (GLP) were distinguished by Smith degradation, methylation analysis, and the wavelength change of the red shift of the mixture of polysaccharide and Congo red in alkaline solution, as well as IR, GC-MS, NMR, and visible spectrometry. The results indicated that GLP was a linear (1→3) β-D-Glcp main chain linkage. Its monosaccharide component was predominantly composed of D-Glc, and small amount of galactose, mannose, xylose and idose, residues of branches terminated with substituted at 1→6 by a small number of single-unit β-D-Glcp side-chains, it′s also observed that the (1→3)-linked β-D- glucan contained a triple-helical conformation, which was composed of a repeating unit with a structure as below:→3)-β-D-Glcp-(1→3)-[β-D-Glcp-(1→3)-]_n-β-D-Glcp-(1→.↑6/1β-D-Glcp
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Objective To observe the counteraction of Ganoderma lucidum polysaccharide(GLP) on the inhibition of mice splenocyte proliferation and IL-1? and IL-2 mRNA expression induced by prostaglandin E2(PGE2).Methods Mixed lymphocyte culture reaction(MLR) method was used for the experiment.The lymphocyte proliferation was determined by MTT method,and the levels of IL-1? and IL-2 mRNA expression were evaluated by semi-quantitative reverse transcriptase polymerase chain reaction(RT-PCR).Results After co-culture with PGE2 for 48 hours,the splenocyte proliferation was inhibited to some extent,and the difference was significant when the concentration of PGE2 was over 10?mol/L(P
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Objective To observe the effects and mechanism of Ganoderma Lucidum Polysaccharides (GLP) on the activity of superoxide dismutase(SOD), the content of malondialdehyde(MDA) of hippocampus tissue and the learning and memory ability. Methods AD rats induced by A? 25-35 were treated with GLP for 7 days and then observing the Morris water maze to study the learning and memory ability, the content of MDA, and the activity of SOD was measured in the hippocampus tissue with spectrophotometer. Ultrastructural changes of neuron were viewed under transmission electron microscope. Results GLP can distinctly improve the learning and memory ability of A?-injected rats(P
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Objective To compare the monosaccharide composition and molar ratio of GL-PPT2, GL-PPT3, and GL-PPT4, which are three polysaccharide peptides isolated from the fruit body of Ganoderma lucidum. Methods The polysaccharide peptides were hydrolyzed by triflouroacetic acid, then derivatized by trichloro-aldehyde-1-pheny-3-methyl-5-pyrazolone (PMP), and determined by HPLC method with the 245 nm detection wavelength. Gradient elution with the solution of KH2PO4 (pH 5.0)-acetonitrile (83.5∶16.5) resulted in baseline separation of nine monosaccharides, of which the chromatographic peaks were very clear. Results GL-PPT2, GL-PPT3, and GL-PPT4 comprise the nine monosaccharides of mannose, ribose, rhamnose, glucose, galactose, arabinose, xylose, galactose, and glucuronic acid. Conclusion The ribose in polysaccharide peptides of G. lucidum is first reported.
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Objective: To study the preventive actions of ganoderma lucidum polysaccharide(GLP) against kidney damage induced by cisplatin. Methods : Female Wistar rats were randomly divided into normal saline(NS) group, cisplatin(CDDP) group, GLP group, CDDP+GLP group. The changes of Scr, BUN, MDA, SOD were measured and renal structure was observed after 5 days by injecting drugs. Results : The contents of serum Scr and BUN of CDDP group were significantly highter than that of NS group. The activity of RBC SOD reduced and the contents of serum MDA increased. The contents of renocortical tissue MDA increased and the activity of SOD declined in renocortical tissue. The contents of serum Scr and BUN of GLP+CDDP group were significantly lower than that of CDDP group. The activity of RBC SOD increased and the contents of serum MDA declined. The concents of renocortical tissue MDA declined and the activity of SOD increased in renocortical tissue. The pathological slice indicated that renal structure was significantly improved. Conclusion : GLP may reduce cisplatin nephrotoxicity and its mechanism may be correlative with that GLP inhibited the blood and renocortical tissue lipid peroxidation increasing.
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Objective To study the effects of complex polysaccharides immune milk (CPIM) on cellular and humoral immune functions in mice. Method Sixty ICR mice were divided into five experimental groups and a control group. The mice in experimental groups were given i.p. Ganoderma lucidum polysaccharide (GLP) immune milk,Lycium barbarum polysaccharides (LBP) immune milk and different doses (low,medium and high) CPIM,and the control group was given equivalently skimmed and sterilized goat's milk respectively for 15 d. The cellular and humoral immune functions and the feces were examined. Results Transformation ability of spleen lymphocytes:There were highly marked differences between all experimental groups and control group,between medium dose group of CPIM and GLB group,LBP group (P