Influence of Ganoderma lucidum spores on the levels of neuropeptide Y and somatostatin in brains of seizure rats

BACKGROUND： Previous research has revealed that somatostatin can induce epilepsy, and that the levels of neuropeptide Y may increase and become more active in brain areas with epileptic seizures. OBJECTIVE： To observe the effect of Ganoderma luciclum spore powder on the neuropeptide Y and somatostatin content in the cerebral cortex and hippocampal regions of seizure rats induced by pentylenetetrazol （PTZ）. Furthermore, to verify any effect of Ganoderma lucidum spore powder on inhibition of epileptic seizures. DESIGN, TIME AND SETTING： A randomized group animal study was performed in August 2007 in the School of Basic Medical Sciences, Jiamusi University （Jiamusi, Heilongjiang, China）. MATERIALS： Thirty healthy, male, Wistar rats, aged 12 weeks and weighing 180-220 g, were taken as the experimental animals. PTZ （Sigma Company, United States） was used to induce epilepsy. Ganoderma lucidum spores （Leyss, ex Fr variety） were purchased from Jiamusi City Wild Growing Case of the Ganoderma Lucidum （China）. Rabbit anti-somatostatin antibodies and secondary antibodies were purchased from Wuhan Boster Company （China）. Neuropeptide Y radioimmunoassay kit was purchased from Beijing Furui Biotechnology Company （China）. METHODS： Thirty rats were randomly divided into three groups： a control group, an epilepsy model group and a Ganoderma lucidum spore-treated group. Each group contained 10 animals. Rats in the epilepsy model group were treated with intraperitoneal injections of PTZ and gastric perfusion of physiologic saline. In the Ganoderma lucidum spore-treated group, intraperitoneal injection of PTZ and gastric perfusion of Ganoderma lucidum spore powder were administered. The blank control group was only administered with the physiological saline by intraperitoneal injection and gastric perfusion. MAIN OUTCOME MEASURES： Immunohistochemical staining and radioimmunoassay methods were used to observe the changes of somatostatin and neuropeptide Y content in brain tissue of epileptic rats, as well as the morphology of neurons. RESULTS： All 30 rats were involved in the result analysis, without any loss. The number of somatostatin immunoreacted positive cells in the cerebral cortex and hippocampus was significantly increased in the epilepsy model group compared to the blank control group （P 〈 0.01）. The number of somatostatin immunoreacted positive cells in cerebral cortex and hippocampus was significantly decreased in the Ganoderma lucidum spore-treated group compared to the epilepsy model group （P 〈 0.01）. The contents of neuropeptide Y in cerebral cortex and hippocampus were significantly increased in the epilepsy model group compared to the blank control group （P 〈 0.01）. The contents of neuropeptide Y in the cerebral cortex and hippocampus were significantly decreased in the Ganoderma lucidum spore-treated group compared to the epilepsy model group （P 〈 0.05）. The epilepsy seizures in the Ganoderma lucidum spore-treated group were obviously reduced compared to the epilepsy model group. CONCLUSION： Ganoderma lucidum spore powder was able to reduce the somatostatin and neuropeptide Y content in the cerebral cortex and hippocampus effectively, so as to achieve an anti-epileptic function and protect neurons from being damaged.

Breaking and Characteristics of Ganoderma Lucidum Spores by High Speed Entrifugai Shearing Pulverizer

The spores of Ganoderma lucidum were ground and broken to ultrafine particles by high speed centrifugal shearing（HSCS） pulverizer. The characteristics of Ganoderma lucidum spores were analyzed by scanning electron microscope （SEM）, Fourier transform infrared spectrophotometry （FTIR）. Ultraviolet-visible pectrophotometer was used to determine the extraction ratio of aqueous solubility polysaccharide between the raw and broken spores. The immunological function on the mice before and after the breaking of spores wan investigated. The experimental results show that after being ground, the sporoderm-broken ratio reachs 100%, the original active ingredients of ganoderma lucidum spores do not change, and the extraction ratio of aqueous solubility polysaccharide is greatly increased by 40.08%. The broken spores show much higher immunological activity comparing with original spores of Ganoderma lucidum.

Ganoderma lucidum spore oil enhances the effect of paclitaxel,improves the tolerance to paclitaxel and prolongs the survival in Lewis tumor-bearing mice

Purpose:This study aims to investigate whether Ganoderma lucidum spore oil(GLSO)could enhance the effect of paclitaxel(PTX),improve the tolerance to PTX and prolong the overall survival of Lewis tumor-bearing mice,which has never been reported before.Methods:The tumor,spleen,and thymus were weighed at the end of the experiment.Whole blood was collected for hematological index analysis,and the intact femur was removed to determine the bone marrow nucleated cell count(BMN).The percentage of lymphocytes in the spleen of mice was detected by flow cytometry,the activity of NK cells was detected by LDH assay,and the proliferation index of lymphocytes was determined by CCK-8 assay.The overall and mean survival time and life extension rate were calculated using SPSS software.Results:Our data showed that GLSO could enhance the anti-tumor effect of PTX and prolong the survival of mice.The underlying mechanisms of the above effects might be related to the toxic reduction effect of GLSO by relieving hematotoxicity,myelosuppression and immunosuppression.Specifically,GLSO could increase the number of blood cells and bone marrow cells,alleviate the thymic index,and elevate the number and activity of NK cells in mice treated with PTX.Conclusion:GLSO may enhance the efficacy of PTX by boosting the activity of immune NK cells and prolong survival by counteracting PTX-induced bone marrow alterations and improving hematopoiesis.These findings suggested the promising role of GLSO in combination with PTX to extend the survival and increase the tolerance of patients in clinical chemotherapy of lung cancer.

Studies on the Triterpenoid Constituents of the Spores from Ganoderma lucidum Karst

Five compounds were isolated from the ether-soluble fraction of the spores of Gano- derma lucidum.On the basis of their chemical properties and spectral data(MS,UV,IR,~1H and ^(13)CNMR),they were identified as 3,7,11,12,15,23-hexaoxo-5α-lanosta-8-en-26-oic acid(Ⅰ),gano- deric acid B(Ⅱ),C(Ⅲ),D(Ⅳ)and ganodermanontriol(Ⅴ).Compound Ⅰ is a new natural product, named ganosporeric acid A.Compounds Ⅱ,Ⅲ,Ⅳ and Ⅴ are known compounds and were obtained for the first time from the spores of Ganoderma lucidum.Pharmacological experiments showed that ganosporeric acid A has an activity for lowering the levels GPT in mice with liver injury by CCl_4 and GaNI and exhibits heptoprotective effects.

Effects of Ganoderma lucidum spores on sialoadenitis of nonobese tiabetic mice

Background Sjǒgren syndrome （SS） is a systematic autoimmune disease, on which traditional therapeutic agents show limited effect. More effective agents with longer-lasting and fewer side effects are needed in the clinic. The aim of this study was to investigate the effects of Ganoderma lucindum spores （GLS） on sialoadenitis of nonobese diabetic （NOD） mice. Methods Thirty-two female NOD mice were assigned randomly into 4 groups： low-dose GLS-treated （L-GLS） group and high-dose GLS-treated （H-GLS） group, a dexamethasone group, and a normal saline （NS） control group. Stimulated total saliva flow rate （STFR）, area of lymphocytic infiltration in submandibular glands and ratios of CD4^＋ and CD8^＋ T lymphocytes and B lymphocytes in peripheral blood as well as apoptosis of these subsets and serum IgG level were tested after 10 weeks of treatments. Differences among the groups were analyzed by one-way analysis of variance （ANOVA）, Student-Newman-Keuls Test （SNK） was used between each two groups and a P 〈0.05 was considered statistically significant. Results STFR of the high-dose GLS group increased significantly after a 10-week treatment compared with those of the NS control group （P 〈0.05）. The incidence of sialoadenitis in GLS-treated NOD mice groups showed no significant difference compared with the control group （P 〉0.05）, but the area of lymphocytic foci in both the H-GLS and L-GLS groups decreased significantly to 50% of the NS control group （P 〈0.05）; the ratio of CD4^＋/CD8^＋ T lymphocytes and apoptosis of B lymphocytes of NOD mice with sialoadenitis were less and apoptosis of CD4^＋ and CD8^＋ T lymphocytes were significantly increased compared with the control group （P 〈0.05）. After pretreatment with H-GLS before sialoadenitis onset, the ratio of CD4^＋/CD8^＋ T lymphocyte and the serum IgG levels of NOD mice decreased significantly （P 〈0.05）. Conclusions Pretreatment with H-GLS can relieve symptoms of sialoadenitis in NOD mice. GLS has some protective effects on sialoadenitis in NOD mice through increasing STFR and decreasing the area of lymphocytic foci by regulating the ratio of CD4^＋/CD8^＋ T and apoptosis of B lymphocytes.

Isolation and Structural Determination of a Glucan from the Spores of Ganoderma lucidum

A water soluble, (1 -->6)-branched, (1 -->4) linked D-glucan (LB-B1), [alpha](D)(21) = +174.2 degrees (c 0.87, H2O), was obtained from a hot-water extract of the sporoderm-broken spores of Ganoderma lucidun (Fr.) Karst by HPSEC, with 0.001 mol/L sodium hydroxide as the eluant, the molecular weight (Mw) of LB-B1 was estimated to be 9.3 x 10(3). From the results of total hydrolysis, methylation analysis, acetolysis and 1D, 2D-NMR experimentation, it was concluded that LB-B1 was composed of repeating units with the following structure: alpha -D-Glc(p)-(1 -->6)-alpha -D-Glc(p)-(1 -->6)-alpha -D-Glc(p) 1 down arrow 6 (-->4)-alpha -D-Glc(p)-(1 -->4)-alpha -D-Glc(p)-(1 -->4)-alpha -D-Glc(p)-(1 -->4)-alpha -D-Glc(p)-(1 -->4-)-alpha -D-Glc(p)-(1 -->)(n)

Magnetic Ganoderma Lucidum Spores(mGLS):A Novel Regulatable Targeted Drug Delivery System

In the past decades,many materials have been studied as carriers for targeted drug delivery.However,there is a need for utilizable and selective carrier materials with few side effects.Here,the magnetic Ganoderma Lucidum Spores(mGLS)as a highly efficient targeted drug delivery carrier were explored.Then the regulatable targeted drug delivery system was verified by loading and releasing of the 5-Fluorouracil(5-FU).The results showed that the maximum of the loaded 5-FU reached 250.23 mg·g^(−1)in the mGLS.The cumulative release of the 5-FU for the drug delivery system could reach 80.11%and 67.14%in the PBS and HCl after 48 h,respectively.In addition,this system showed the good pharmacokinetic properties in vivo.After 12 h,the blood concentration in the 5-FU@mGLS group kept at 5.3µg·mL^(−1)and was four times higher than that in the 5-FU group.In summary,the GLS as a natural microscale core-shell structures appears the striking application in carrier material for oral drug delivery.

Aqueous-soluble components of sporoderm-removed Ganoderma lucidum spore powder promote ferroptosis in oral squamous cell carcinoma

Objective: Ferroptosis is a novel cell death process which displays a promising role in cancer treatment.However, clinically available drugs targeting ferroptosis are rarely used, and yet there are no studies reporting on inducing ferroptosis via Chinese herbal extracts. Here we explored the tumor inhibition effects of Ganoderma lucidum(G. lucidum) on oral squamous cell carcinoma(OSCC). Specifically, we aimed to clarify the biological mechanism of components in the dietary, aqueous-soluble sporoderm-removed G. lucidum spore powder(A-GSP).Methods: Preliminary transcriptome analysis revealed the significant enrichment of the ferroptosis pathway.Cellular Fe2+, glutathione(GSH), malondialdehyde(MDA), reactive oxygen species(ROS) and lipid peroxide levels were measured to identify ferroptosis occurrence. Western blotting was used to measure ferroptosis-related proteins. Changes in mitochondria morphology and function were observed with transmission electron microscopy(TEM) and ATP detection assays. Ferroptosis inhibitor ferrostatin-1 was then used to verify the anti-tumor effects of A-GSP. Finally, nude mice xenograft models of oral cancer confirmed that A-GSP inhibited tumor growth.Results: A-GSP promoted ferroptosis in oral cancer cells by inducing Fe2+influx, GSH depletion, as well as lipid peroxide and ROS accumulation. Ferroptosis-related proteins exhibited corresponding changes, particularly Acylco A synthetase long chain family member 4(ACSL4) increase and glutathione peroxidase 4(GPX4) decrease. A-GSP considerably lowered mitochondrial volume and ridge number, while significantly decreasing ATP production. Ferrostatin-1 reversed all of these A-GSP-induced changes. In vivo, A-GSP exerted a ferroptosismediated tumor-suppressing effect without observable adverse reactions.Conclusions: Our findings demonstrate the therapeutic potential of A-GSP for treating patients with OSCC by targeting ferroptosis.

Volatile Components and Antitumor Activity of Ganoderma lucidum Spore Oil and Its Molecular Distillation Components

[Objectives]To compare the effects of molecular distillation on the flavor and antitumor activity of Ganoderma lucidum spore oil.[Methods]G.lucidum spore oil was separated and purified by molecular distillation technology,and the volatile components of different components of molecular distillation were analyzed by gas chromatography-ion mobility spectrometry(GC-IMS)technology.Human liver carcinoma cells(HepG2),human breast cancer cells(MCF-7),and human cervical cancer cells(Hela)were selected as the tumor cell lines to be tested,and the cell viability was detected by the MTT assay.[Results]Molecular distillation effectively reduced small molecular substances produced by oil oxidation in G.lucidum spore oil,such as heptanal,octanal,linalool,hexanal,E-2-octanal,3-ethylpyridine,etc.Among the heavy components,the content of esters was relatively high,mainly including ethyl levulinate,ethyl crotonate,and amyl butyrate.The MTT cytotoxicity test indicated that G.lucidum spore oil and its molecular distillation components had certain inhibitory effects on the growth of three tumor cells,and G.lucidum spore oil crude oil had the most significant antitumor activity.G.lucidum spore oil crude oil,heavy component,and light component had the most significant antitumor activity on HepG2 cells,followed by MCF-7 cells,and the weakest antitumor activity on Hela cells.The quality of G.lucidum spore oil became higher after molecular distillation,and the rancid smell was reduced,and molecular distillation had little effect on the antitumor activity of G.lucidum spores.[Conclusions]Molecular distillation technology can be applied to the refining of G.lucidum spore oil to improve product quality.

Antitumor Activity of the Ganoderma Lucidum Spore Alcohol Extract in Vitro

Several cancer cell lines(epithelioma cells or leukemia cells)from human being or mouse were first used to study the antitumor activity of the Ganoderma lucidum spore alcohol extract(GLSAE)in vitro by the MTT test A comparision was made between the sporodermbroken(SB)and sporoderm nonbroken(SN)GLSAE It was showed that both GLSAE SB and GLSAE SN could inhibit the proliferation of these cancer cells,but the activity of GLSAE SB was much higher than that of GLSAE SN These results suggested that Ganoderma lucidum spore could probably be used for tumor treatment

Effect of Ganoderma lucidum spore on expression of insulin-like growth factor-1, nuclear factor-kappa B, and neuronal apoptosis in the epileptic rat brain

BACKGROUND：It has been reported that Ganoderma lucidum spore powder, a very well known Chinese traditional medicine, can affect immunoregulation, free radical scavenging, and anti-hypoxia responses. OBJECTIVE： To investigate the effect of Ganoderma lucidum spore powder on expression of insulin-like growth factor-1 （IGF-1）, nuclear factor-κB （NF-κB） and neuronal apoptosis in rats with pentylenetetrazol （PTZ）-induced epilepsy. DESIGN, TIME AND SETTING： A cellular and molecular biology experiment with randomized controlled study design was performed at the Central Laboratory of Basic Medical College of Jiamusi University from June to August 2005. MATERIALS： Thirty healthy, adult, male, Wistar rats were selected and randomly divided into 3 groups （10 rats per group）： control, epilepsy model, and Ganoderma lucidum spore powder. A sub-eclampsia PTZ dose （35 mg/kg） was intraperitoneally injected to induce epilepsy in the latter two groups. Wild Ganoderma lucidum spore powder （30 g/L） was provided by the wild Ganoderma lucidum plant nursery at Jiamusi, China. Immunohistochemical detection and terminal deoxynucleotidyl transferase-mediate dUTP nick end-labeling （TUNEL） kits were purchased from Wuhan Boster Biological Technology Co., Ltd., China. METHODS： Ganoderma lucidum spore powder was intragastrically administered at a dose of 10.0 mL/kg, once a day for 28 days. In the epilepsy and control groups, an equivalent volume of normal saline was intragastrically administered. MAIN OUTCOME MEASURES： Immunoreactivity for IGF-1 and NF-κB/P65 were detected by immunohistochemical staining. Neuronal apoptosis was detected using TUNEL methods. RESULTS： The hippocampus and cerebral cortex of rats with PTZ-induced epilepsy exhibited a higher number of apoptotic cells at high magnification （×400）, compared with the control group. Expression of IGF-1 and NF-κB were higher in the epilepsy group, compared with the control group （P 〈 0.01）. In Ganoderma lucidum spore-treated rats, fewer apoptotic cells were observed in the hippocampus and cerebral cortex, expression of NF-κB/P65 was lower, and immunoreactivity to IGF-1 increased more distinctly, compared with the epilepsy group. In addition, seizure latency was longer on 17, 21, and 25 days post-PTZ treatment in the Ganoderma lucidum spore powder group, compared with the epilepsy group （P 〈 0.05-0.01）. CONCLUSION： Ganoderma lucidum spore powder down-regulated expression of NF-κB in brain tissues of rats with PTZ-induced epilepsy, increased immunoreactivity to IGF-1, and inhibited neuronal apoptosis. These results indicated that Ganoderma lucidum spore powder has a neuroprotective effect.

Ganoderma lucidum spore oil(GLSO),a novel antioxidant,extends the average life span in Drosophila melanogaster

In ancient China,Ganoderma lucidum was believed to be a medical fungus that could increase lifespan.Recently,pharmacologic studies have found that polysaccharide peptides and triterpenoids extracted from Ganoderma lucidum have various physiological effects as active compounds.However,the effects of spore oil isolated from Ganoderma lucidum remains unknown.In this study,the biological effects of Ganoderma lucidum spore oil(GLSO)were evaluated using a Drosophila melanogaster model.Compared with untreated groups,groups treated with GLSO had significantly longer average and maximum lifespan in both normal conditions and under oxidative stress.The activities of various antioxidant enzymes were measured to determine the antioxidant effect of GLSO.GLSO treatment markedly enhanced total superoxide dismutase(SOD)and catalase(CAT)activity and decreased levels of malondialdehyde(MDA).Further,we found dose-dependent increases in the mRNA expression of Cu,Zn-SOD,Mn-SOD,and CAT in GLSO-treated groups.These results suggest that GLSO may effectively eliminate free radicals and extend lifespan in Drosophila.Future work should investigate the value of GLSO as a functional food for the prevention of aging in larger animal models.

Ganoderma lucidum spore powder modulates Bcl-2 and Bax expression in the hippocampus and cerebral cortex,and improves learning and memory in pentylenetetrazole-kindled rats

We studied the effects of Ganoderma lucidum spore powder on Bax and Bcl-2 expression and neuronal apoptosis in pentylenetetrazole-kindled epileptic rats. Sixty adult rats were randomly divided into a control group, an epileptic group （kindled） and three medication groups （150,300 450 mg/kg given to kindled rats）. Bax and Bcl-2 immunohistochemistry and TUNEL labeling showed that the number of Bax- and TUNEL-positive cells in the hippocampus and cerebral cortex decreased significantly in the high-dose medication group, while the number of Bcl-2 immunoreactive cells increased. The Morris water maze test showed that high-dose treatment significantly shortened escape latency and increased spatial probe trial performance. Our findings indicate that a high dose of Ganoderma lucidum spore powder upregulates the expression of antiapoptotic Bcl-2 protein in the hippocampus and cerebral cortex, inhibits proapoptotic Bax expression, and decreases seizure-induced neuronal apoptosis. Further, Ganoderma lucidum appears to protect against epilepsy-related learning and memory impairments.

Effects of Ganoderma lucidum spore powder on astrocyte expression and glutamine synthetase activity in the hippocampal region of epileptic rats

BACKGROUND： Recent studies have demonstrated that astrocyte dysfunction plays a central role in inhibiting epileptic seizures and that regulation of astrocyte function may be a new target for treatment of epilepsy. OBJECTIVE： To observe the effects of Ganoderma lucidum spore powder （GLSP） on astrocyte morphology and glutamine synthetase （COS） activity in the hippocampal region of epileptic rats. DESIGN, TIME AND SETTING： A randomized, controlled animal experiment was performed at the Function Laboratory, College of Basic Medicine, Jiamusi University between October and December 2006. MATERIALS： A total of 30 Sprague Dawley （SD） rats were randomized to three groups （n = 10）： control, model, and GLSP. GLSP was sourced from Jiamusi Wild Ganoderma Lucidum Planting Base and prepared to 30 g/L with physiological saline before use. Pentylenetetrazol （PTZ） （10 g/L） was provided by Sigma Company, USA. METHODS： The control group received intraperitoneal （i.p.） and intragastric （i.g.） physiological saline. Following epilepsy induction by i.p. administration of PTZ （35 mg/kg）, rats from the mode/and GLSP groups were ig injected with physiological saline and GLSP （300 mg/kg）, respectively. Each compound was administered once per day, for a total of 28 successive days. Epileptic seizure convulsions were graded 0-5. A higher grade indicated more severe epilepsy. Only those rats showing stage 2 or higher convulsions at least 5 times successively were included in further experiments. MAIN OUTCOME MEASURES： Immediately alter injection, seizure activity was monitored for 30 minutes to determine the latent period and seizure duration; simultaneously, astrocyte numbers and GS activity in the hippocampal region of rats with epilepsy were detected by immunohistochemistry. RESULTS： All 30 rats were included in the final analysis. On day 28, following PTZ administration epileptic seizures were not found in the control group. In the GLSP group, rats exhibited rhythmic head nodding or facial spasms, and the latent period was significantly longer than that of the model group （P 〈 0.05）. In the model group, the majority of rats exhibited myoclonus or generalized convulsions, and epileptic seizure duration was slightly （but not significantly） longer than that in the GLSP group （P 〉 0.05）. Within 2-3 minutes of PTZ injection, the model group exhibited grade 4 or 5 epileptic seizures, and the GLSP group mostly showed grade 2 or 3, and occasionally grade 4 or 5, epileptic seizures. All rats recovered within 30 minutes. The model group exhibited significantly increased astrocytes （P 〈 0.05）, with thicker cellular processes and a higher number of cellular processes, and significantly decreased GS activity （P 〈 0.05） tban the control group. The astrocyte count was significantly decreased but GS activity was significantly increased in the GLSP group when compared with the model group （P 〈 0.05）; however, astrocyte appearance was similar in both groups （P 〈 0.05）. CONCLUSION： GLSP can effectively inhibit astrocyte numbers and elevate GS activity in the hippocampal region of rats with epilepsy, thereby reducing epileptic seizures.

Study on Key Techniques for High-quality and High-yield Cultivation of Ganoderma lucidum Spore Powder

Comparative test of 4 Ganoderma lucidum varieties from different sources showed that the mycelium of Chizhi 1 grew fast with thick and dense hyphae,round and solid cap,and high spore powder yield.Chizhi 1 was proved to be an excellent variety because of its strong resistance and high spore powder yield.Different cultivation materials were chosen and combined to form 3 cultivation formulations.The results showed that Formulation(3),in which basswood was soaked in nutrient solution for 24 h,presented fast mycelial growth and high spore powder output,and therefore was proved to be a high-yield formulation of Ganoderma lucidum spore powder.

Bioactive components,pharmacological properties and underlying mechanism of Ganoderma lucidum spore oil:A review

Ganoderma lucidum is a Chinese medicinal fungus with a long history of use in healthcare and disease treatment.G.lucidum spores(GLS)are tiny germ cells released from the mushroom cap during the mature stage of growth.They contain all the genetic active substances of G.lucidum.G.lucidum spore oil(GLSO)is a lipid component extracted from broken-walled Ganoderma spores using supercritical CO_(2)extraction technology.GLSO contains fatty acids,Ganoderma triterpenes,sterols and other bioactive compounds.Previous studies have demonstrated that GLSO has a wide range of pharmacological properties,including anti-tumor,anti-aging,neuroprotection,immunomodulation,hepatoprotection and modulation of metabolic diseases.This review summarizes the research progress of GLSO over the past two decades in terms of its bioactive components,extraction and processing techniques,pharmacological effects and safety evaluation.This provides a solid foundation for further research and application of GLSO.

Antitumor effects of different Ganoderma lucidum spore powder in cell-and zebrafish-based bioassays

Objective:Ganoderma lucidum spore(GLS)is gaining recognition as a medicinal part of G.lucidum and has been reported to possess various pharmacological properties,such as antitumor activity.In this work,wall-broken GLS powder(BGLSP)and wall-removed GLS powder(RGLSP),two kinds of GLS powder with different manufacturing techniques,were compared in terms of contents of active constituents and in vivo and in vitro antitumor effects.Methods:The ultraviolet and visible spectrophotometry method was used to determine the contents of polysaccharides and total triterpenoids in BGLSP and RGLSP.Seventeen individual triterpenoids were further quantified using ultra-high-performance liquid chromatography and quantitative analysis of multicomponents by single marker.The antitumor effects of BGLSP and RGLSP were evaluated using in vitro cell viability assay against human gastric carcinoma SGC-7901,lung carcinoma A549 and lymphoma Ramos and further validated by in vivo zebrafish xenograft models with transplanted SGC-7901,A549 and Ramos,Results:The results showed that the contents of polysaccharides,total triterpenoids and individual triterpenoids of RGLSP were significantly higher than those of BGLSP.Although both BGLSP and RGLSP inhibited the three tumor cell lines in vitro in a dose-dependent manner,the inhibitory effects of RGLSP were much better than those of BGLSP.In the in vivo zebra fish assay,RGLSP exhibited more potent inhibitory activities against tumors transpla nted into the zebra fish compared with BGLSP,and the inhibition rates of RGLSP reached approximately 78%,31%and 83%on SGC-7901,A549 and Ramos,respectively.Conclusion:The results indicated that the antitumor effects of GLS were positively correlated with the contents of the polysaccha rides and triterpenoids and demonstrated that the wall-removing manufacturing technique could significantly improve the levels of active constituents,and thereby enhance the antitumor activity.