Preparation of disulfiram naonosuspensions and their anti-tumor efficacy in vitro and in vivo / 药学学报

Disulfiram (DSF) is a traditional anti-alcohol drug, but it was recently found that DSF has strong inhibitory effect on the growth of a variety of cancer cells. However, its clinical application is greatly limited due to its poor solubility, instability in gastrointestinal tract and short plasma half-life. In this study, DSF is fabricated into nanosuspensions with the aim of trying to solve these problems. DSF nanosuspensions (DSF-NSps) were prepared by the anti-solvent precipitation method under ultrasonication, and the suitable stabilizer was screened according to the size, polydispersity index (PDI), and zeta potential of the resultant nanosuspensions, along with their particle size change during the storage at room temperature. The particle size, PDI, and zeta potential of DSF-NSps were determined using dynamic light scattering method, while the morphology of DSF-NSps was observed by transmission electronic microscope (TEM). The stability of DSF-NSps in media was examined according to their particle size change in different physiological media. The concentration of DSF was measured by HPLC assay. The in vitro drug release was evaluated on basis of dialysis. MTT assay was employed to evaluate the in vitro cytotoxicity of DSF-NSps against cancer cell lines. The 4T1 tumor-bearing mouse model was used to evaluate the in vivo therapeutic efficacy of DSF-NSps. All the animal experiments were acquired according to the Regulations for Animal Experiments and Guidelines for Ethical as defined by Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College. As a result, the combinational use of soyabean lecithin (SPC) and D-alpha tocopherol acid polyethyene glycol succinate (TPGS) was determined to best stabilize DSF-NSps when the ratio of DSF-SPC-TPGS was 24∶20∶4 (weight ratio), with small particle size and good storage stability. The resultant DSF-NSps showed a regular spherical morphology and drug loading content of (45.36 ± 2.09) %, with average particle size of 175.00 ± 0.75 nm, PDI of 0.24 ± 0.07 and zeta potential of -14.3 mV. DSF-NSps displayed good particle size stability in a variety of biological media including phosphate buffer saline, normal saline, 5% glucose, artificial gastric fluid, artificial intestinal fluid and plasma, which would meet the demand of both intravenous and oral administration. The in vitro study demonstrated that nano-encapsulation greatly increased the stability of DSF in aqueous media, DSF-NSps exhibited sustained release of the encapsulated drug and significantly inhibited 4T1 cells compared to free DSF (IC50, 1.07 vs 5.53 μg·mL-1, P<0.01). DSF-NSps showed a good dose-response relationship on the 4T1 tumor-bearing mice with the tumor inhibition rates at the three doses being 80.22%, 75.14% and 66.10%, all higher than that of paclitaxel injections (55.01%, P<0.05). The in vivo biodistribution study displayed that DSF-NSps were mainly distributed into liver, spleen and tumor. In sum, disulfiram nanoparticles could be expected to provide an effective anti-cancer drug for the treatment of breast cancer.

Protective effect of raspberry extract on ConA-induced acute liver injuryin mice / 中国中药杂志

In this experiment,the antioxidant capacity of raspberry extract and the protective effect on liver injury induced by ConA in mice were investigated. Balb/C male mice were randomly divided into six groups: normal group,model group,bicyclol control group( 200 mg·kg~(-1)),low-dose raspberry extract group( 200 mg·kg~(-1)),middle-dose raspberry extract group( 400 mg·kg~(-1)),and highdose raspberry extract group( 800 mg·kg~(-1)). Each group was intragastrically administered with drugs according to the body weight once a day. Seven days later,all of the groups except for the normal group were treated with ConA( 20 mg·kg~(-1)) through tail vein injection to establish the acute liver injury model. The mice were put to death 8 hours later. The organ indexes were calculated. These rum levels of ALT,AST and LDH and the activities of SOD,CAT,GSH and MDA in liver tissue were detected. HE staining was used to observe the pathological changes of liver tissue in mice. Western blot was used to detect the expressions of Bax,Bcl-2,Nrf2 and Keap-1. The antioxidant capacity of raspberry extract was measured by CAA assay. The results showed that,raspberry extract had a strong antioxidant capacity. Simultaneously,compared with the model group,raspberry extract can significantly improve the pathological conditions of liver,and significantly reduce ALT,AST and LDH activities in serum of liver injury mice( P<0. 01). The activities of SOD,CAT in liver homogenate supernatant were significantly increased in the high-dose group,the content of GSH increased,while the content of MDA was sharply declined in the high-dose group( P<0. 01). Meanwhile,raspberry extract down-regulated the expressions of Bax and Keap-1 and up-regulated the expressions of Bcl-2 and Nrf2. CAA showed that the compound raspberry extract had a strong antioxidant capacity. Therefore,raspberry extract has an obvious protective effect on acute liver injury induced by ConA in mice.

Protective effect of scutellarin on myocardial ischemia/reperfusion injury and its mechanism / 中国药理学通报

Aim To study the protective effect of scutellarin (Scu) on ischemia/reperfusion (I/R) injury in isolated rat hearts and its mechanism. Methods Acute myocardial I/R injury was induced by Lange-ndorff perfusion in isolated rat hearts. SD rats were randomly divided into five groups; blank group, model group, low, medium and high dose group of Scu (0.3, 3, 30 mg • kg-1). The drug experimental group was perfused with K-H solution containing Scu (0. 3, 3, 30 mg • L-1) in vitro perfusion. Myocardial contractile function was recorded; AST, CK and LDH activities in rat myocardium were detected by kit; ICAM-1, IL-ip, IL-6, IL-18 and TNF-α were detected by ELISA; the morphological changes of myocardium were observed by HE staining; and the expressions of IL-1 p, NLRP3 and NF-kB were detected by Western blot. Results Compared with blank group, the contractile function of isolated rat heart in model group significantly decreased, the activities of AST, CK and LDH in myocardium significantly increased, and the contents of inflammatory factors such as ICAM-1, IL-lp, IL-6, IL-18 and TNF-a significantly increased. The expression of IL-1 (3 and NLRP3 and the nuclear translocation of NF-kB significantly increased. Scu significantly improved the above pathological changes. Conclusion The protective effect of Scu on myocardial I/R injury may be related to the inhibition of NF-KB/NLRP3/IL-1 p pathway.

Effects of cultivated Cordyceps sinensis on proliferation and migration of B16 melanoma cells / 中草药

Objective To investigate the effects of cultivated Cordyceps sinensis on inhibiting the proliferation and migration of B16 melanoma cells. Methods MTT assay and clone formation assay were used to examine the inhibitory effect of cultivated C. sinensis on the proliferation of B16 cells; Scratching test and transwell assay were used to detect its effect on migration; Cell adhesion assay was used to detect its effect on adhesion. Flow cytometry was used to detect its effect on cell cycle arrest; Western blotting was used to detect its effect on the expression of the proteins of MMP-2, MMP-9, Bax, Bcl-2, CyclinD1, CDK2, CDK4, P21, and p-Akt. Results The results showed that cultivated C. sinensis can significantly inhibit the proliferation and clone formation of B16 cells via arresting cells at G1/S phase in a dose-dependent manner. Moreover, cell migration was also substantially inhibited in a dose-dependent manner. Western blotting analysis showed that cultivated C. sinensis obviously increased the levels of Bax and P21, and meanwhile, decreased the expression of Bcl-2, MMP-2, MMP-9, CDK2, CDK4, CyclinD1, and p-Akt. Conclusion The inhibitory effects of cultivated C. Sinensis on the proliferation and migration of B16 cells probably associated with the expression of related regulating proteins, MMPs family proteins and p-Akt protein in cell cycle.

Structural modification and anti-inflammatory biological evaluation of monomeric compounds from Aralia elata / 中草药

Objective: Based on structural modification of monomeric compound calenduloside E from Aralia elata, to evaluate anti-inflammatory activity of the analogues. Methods: Applying oleanolic acid as starting material, the target compounds were prepared by seven steps reactions and evaluated for anti-inflammatory effects by RAW264. 7 cells in vitro. Results: Ten analogues G1-G5 and H1-H5 were synthesized. The structures of the target compounds were identified by spectrum. Pharmacological results showed that all of the compounds had different levels potency of anti-inflammatory effects in cells. In particular, compounds G1-G4 and H1-H3 showed significant anti-inflammatory activity comparing wiht lead compounds. Conclusion: The new compounds G1-G5 and H1-H5 which showed potential of anti-inflammatory biological activity, had not been reported in any literatures and deserved further research.

Preparation of annonaceous acetogenins nanosuspensions using poloxamer 188 as a stabilizer and their in vitro and in vivo investigation / 药学学报

Annonaceous acetogenins (ACGs) are effective part extracted and separated from Annona squamosa seeds, they have good antitumor activity against a variety of tumor cells. However, the solubility of ACGs is poor with serious toxic and side effects, which greatly limits their application in clinical practice. In this study poloxamer 188 (P188) was selected as a drug carrier or a stabilizer to prepare ACGs nanosuspensions (ACGs-NSps) using anti-solvent precipitation. The nanosuspensions were examined via dynamic light scattering (DLS) method to examine size of the nanosuspensions. Transmission electron microscopy was used to observe their morphology. HPLC assay was used to measure their drug loading content and the in vitro drug release. The stability of ACGs-NSps at room temperature, in various physiological media and plasma, and the hemolytic test and lyophilization were all investigated. MTT assay was performed to study the cytotoxocity of ACGs-NSps against four tumor cell lines. 4T1 bearing tumor model was used to assess their in vivo antitumor therapeutic efficacy. The obtained ACGs-NSps were spherical, the average particle size was 169.4±1.25 nm, the polydispersity index (PDI) value was 0.130±0.020, the zeta potential was -19.8 mV and the drug loading content was 48.18%. ACGs-NSps were stable at room temperature for at least 15 days. They could be lyophilized in the presence of 0.5% glucose and 2.0% P188. ACGs-NSps showed sustained in vitro drug release, and the cumulative drug release reached 80.82% within 144 hours. ACGs-NSps maintained their particle size in various physiological media, and plasma with no hemolysis and then met demands of both oral and intravenous administration. In contrast to free ACGs, ACGs-NSps displayed significantly higher cytotoxicity against 4T1 (IC50, 0.892±0.124 μg·mL-1 vs 2.495±0.108 μg·mL-1, P 50, 0.747±0.051 μg·mL-1 vs 2.204±0.064 μg·mL-1, P 50, 2.265±0.081 μg·mL-1 vs 4.159±0.071 μg·mL-1, P 50, 0.473±0.024 μg·mL-1 vs 1.196±0.022 μg·mL-1, P in vivo study demonstrated that the daily oral administration of ACGs-NSps (3 mg·kg-1) resulted in higher tumor inhibition rate compared to ACGs/oil solution (67.23% vs 53.11%), comparable to the intravenous injection of 0.5 mg·kg-1 ACGs-NSps every other day (70.34%). Nanosuspensions effectively solved the problem of ACGs insolubility and difficulty in drug delivery. Using P188, a pharmaceutic adjuvant approved by FDA for iv injection, the resultant ACGs-NSps appear promising as an anti-tumor drug that can be used in clinic.

Preparation of gambogic acid naonosuspensions and their anti-tumor efficacy / 药学学报

Gambogic acid (GA), the main active ingredient in gamboge, has been reported to have good anti-tumor activity with excellent selectivity. However, its clinical application is limited by the poor water solubility. GA nanosuspensions were designed in this study in order to solve this problem. GA nanosuspensions were prepared by microprecipitation method based on pH adjustment. Suitable stabilizer was screened according to the size and polydispersity index (PDI) of the resultant nanosuspensions. Dynamic light scattering method was used to measure the particle size and transmission electron microscopy was used to observe the morphology. The stability was studied in different medium. The drug release was evaluated using a dialysis method. MTT assay was used to assess their cytotoxicity in vitro against cancer cell line. Anti-tumor effect in vivo was investigated on H22-bearing mice. In result, Poloxamer (P188) was found to be a good stabilizer. The resultant GA nanosuspensions (GA-NSps) were 135.9 ±5.1 nm in diameter, with PDI value being 0.26 ±0.01 and the zeta potential being −35.1 ±1.36) mV. GA-NSps were nearly spherical. They were quite stable in various physiological media. GA-NSps exhibited a sustained drug release pattern, with the cumulative release reaching 90.26% within 312 h. In MTT assay, GA-NSps had a stronger cytotoxicity against HepG2 cells than the free drug (IC50, 0.851 8 μg·mL−1 vs 2.104 μg·mL−1, P vs 66.80%, P < 0.01). In summary, we prepared GA-NSps with high drug loading capacity, small particle size and good stability, and provided a solid basis for the effective dosage form of gambogic acid.

Honokiol nanosuspensions:preparation, in vitro and in vivo evaluation / 药学学报

Honokiol (HK) have extensive pharmacological activities, but its poor solubility and instability restricted its clinical application and efficacy exertion. HK nanosuspensions (HK-NSps) were designed in this study in order to solve the problems. HK-NSps were prepared by antisolvent precipitation method, using poly-vinylpyrrolidone (PVP) and bovine serum albumin (BSA) as a combined stabilizer. The particle size was measured using dynamic light scattering method, the morphology was observed by transmission electron microscopy. The size change and drug content of HK-NSps in various physiological media during the storage at ambient temperature was examined to evaluate their storage stability. Dialysis method was used to study their drug release in vitro. MTT assay was used to assess their in vitro cytotoxicity against 4T1 breast cancer cell line. Anti-tumor effect in vivo was also investigated in 4T1 tumor-bearing mice. HK-NSps were prepared with high drug loading content of 48.62%, nearly spherical shape and good storage stability. The average particle size was (83.40 ±1.042) nm, the polydispersity index (PDI) value was 0.223 ±0.011, the zeta potential was (-42.2 ±1.2) mV. HK-NSps showed sustained in vitro drug release and enhanced cytotoxicity in contrast to free HK against 4T1 cells (IC50, 8.36 μg·mL-1 vs 37.58 μg·mL-1, Pin vivo study on 4T1 tumor-bearing mice demonstrated that HK-NSps showed good dose-dependent tumor inhibition rate (TIR). In contrast to 4 mg·kg-1 of PTX injection (TIR, 47.9%), medium and high dose of HK-NSps displayed improved therapeutic efficacy (TIR, 55.67% for 40 mg·kg-1, 67.28% for 60 mg·kg-1, P-1) had TIR of only 54.13% even administrated every day. In conclusion, HK-NSps were prepared with small size, high drug-loading capacity, and good stability. The improved in vitro and in vivo antitumor efficacy demonstrated that HK can be a promising antitumor drug in combination with nanosuspensions technology.

Synthesis and lipid-regulating evaluation of caffeic acid amide derivatives / 中草药

Objective To evaluate the anti-lipid metabolic disorder activities of these compounds in vitro, the natural caffeic acid amide derivatives were designed and synthesized. Methods Using caffeic acid as start material, BOP as a condensing agent, and the target compounds were sequentially prepared with fourteen amines, and then the lipid-regulating effect was evaluated using HepG2 cells. Results Fourteen caffeic acid amide compounds CA1-CA14 were synthesized. The structures of the target compounds were identified by spectrum. Pharmacological results showed that fourteen derivatives have potency of lipid-regulating in different levels. In particular, compound CA6 showed significant lipid-regulating effects compared to the lead compounds caffeic acid and Simvastatin. Conclusion Compound CA6, CA7, and CA11 had not been reported in any literatures before. Among them, the novel compounds CA6 and CA11 have showed potential of lipid-regulating activities, and deserved further research.

Research progress on pharmacological effects of sophocarpine / 中草药

Sophocarpine is an alkaloid extracted from the dry roots and ground parts of Leguminous Sophora flavescens or S. alopecuroides. Sophocarpine has a wide range of sources, it is abundant in S. flavescens and S. alopecuroides, and it is easy to extract. Sophocarpine has a wide range of pharmacological effects, such as antitumor, anti-inflammatory and analgesia, antivirus, liver protection, cardiovascular protection and so on. In this paper, the pharmacological effects of sophocarpine were reviewed in order to provide references for the treatment of related diseases and new drug research and development.

Trace phenolic compounds from Red Yeast Rice / 中国中药杂志

Trace chemical constituents from the ethyl acetate extract of Red Yeast Rice were investigated. Four phenolic compounds were isolated by various column chromatographies, and their structures were identified on the basis of spectroscopic analysis including UV, MS, IR and NMR. The four compounds were identified as 2-methyl-5-(2'R-methyl-4'-hydroxy-butyl)-cinnamic acid(1), 5-(2'-hydroxy-6'-methyl phenyl)-3-methylfuran-2-carboxylic acid(2), daidzein(3), and genistein(4). Compound 1 was new and 2 was firstly discovered from the genus Monascus, while 3-4 were obtained from Red Yeast Rice for the first time.

Study on mechanism of HepaRG cell injury induced by bavachinin / 中国药理学通报

Aim To probe into the bavachinin toxic mechanisms in HepaRG cells. Methods MTT assay was used to determine toxicity and dose. After treated with 6.25,12.5 and 25 μmol·L-1bavachinin for 24 h,the cell death was investigated by Hoechst 33342/PI,LDH,caspase-3 activity,and Bax,Bcl-2 expression levels. Mitochondrial damage was detected by mito-chondrial membrane potential,ATP content,openness of mitochondrial permeability transition pore and cyto-chrome C level. Results Bavachinin administration up-regulated Bax/Bcl-2 ratio and caspase-3 activity with increase of drug concentration after 24 h. Apop-totic rate increased in a dose-dependent manner be-tween 6.25 and 25 μmol·L-1,but when reached to 25 μmol·L-1,cells showed more necrosis. Mitochon-drial injury indicators significantly increased after 24h. Conclusion Bavachinin induces HepaRG cell apopto-sis and necrosis through mitochondria injury.

Research progress on pharmacological effects of tetrahydroberberine / 中草药

Tetrahydroberberine (THB) belongs to the alkaloid of tetrahydroisoquinoline, which is derived from the roots of Corydalis yanhusuo and can also be hydrogenated from berberine. THB has a variety of significant biological activities compared to berberine. It is reported that THB has the effects of anti-hypertension, anti-arrhythmia, anti-fibrillation and against acute myocardial infarction, and also can treat and protect the injury of ischemic and reperfusion. Moreover, other research has found its effects upon anti-oxidant and regulating the functions of gastrointestinal tract. By searching literature of domestic and foreign from Pubmed, CNKI and other databases, pharmacological activities of THB were summarized in this paper, in order to provide reference for the further study of THB.

Research progress on chemical constituents and pharmacological effects of alkaloids from plants of Lycoris Herb / 中草药

Recently, studies on Lycoris Herb. type alkaloids received the attention of scholars home and abroad. Lycoris Herb. type alkaloids can be devided into seven types according to their molecular structure, including lycorine type, crinine type, galanthamine type, tazettine type, narciclasine type, lycorenine type, homolycorine type, and montanine type. Researches have shown that Lycoris Herb. type alkaloids possess multiple phamocology activities, such as strong antitumor activity on human breast cancer cell (MCF-7), human leukemia cell (HL-60); and strong inhibitory effect of flu virus, measles virus, polio virus, and SARS virus; Lycorine type and crinine type alkaloids have antimalaria effect; Besides, lycorine type has strong anti-acetylcholinesterase effect. In a word, lycorine type and lycoris type alkaloids carry multiple pharmacological effect and belong to promising substances.

Impact of AMPKgamma silencing on AMPK activation and intracellular lipids regulation / 药学学报

The study is aimed to confirm the silencing efficiency of the vector in human hepatocellular liver carcinoma cell line (HepG2), and observe effects of AMPKgamma silencing on the AMPK stimulating activity and lipid synthesis of cordycepin (CCS), a natural product with known AMPK activating function. The downregulating efficacy of siRNAs on AMPKgamma expression was confirmed in our previous study. The double stranded shRNA Oligo was ligated to lentivirus vector and verified by sequencing. The lentiviral which can effectively inhibited protein expression levels of AMPKgamma was selected by Western blotting, and the regulation of CCS on protein expression of AMPKgamma and p-AMPK in AMPKgamma silence cells were detected by Western blotting analysis. The lipid accumulation in cells was observed by Oil-Red O stain and cells were collected for the estimation of cholesterol (TC), triglyceride (TG). The results showed that the lentiviral vector carrying a shRNA targeting the AMPKgamma gene was successfully constructed. Western blotting analysis confirmed that GR085 had the highest interfering efficiency. Treatment with CCS can significantly increase the levels of phospho-AMPK in normal cells, and the level of TC, TG was reduced, but in AMPKgamma silence cells the effects of CCS on AMPK activation and lipid synthesis were almost completely abolished without changing the expression levels of total AMPK or AMPKgamma protein. In conclusion, the AMPKgamma gene may be related to AMPK activation and intracellular lipids regulation by CCS.

Research progress of transgenic Drosophila model of Alzheimer disease / 药学学报

Alzheimer disease (AD) is a common neurodegenerative disease. Drosophila has been regard as one of the ideal models for Alzheimer because of its unique advantage on genetic manipulation. AD transgenic drosophila models not only help to elucidate the pathogenesis of Alzheimer disease, but also provide potential screening models for drugs to treat the disease. In this review, we summarize the recent research progress using AD transgenic drosophila.

Effects of solanine on microtubules system of human breast cancer MCF-7 cells / 中草药

Objective To investigate the effects of solanine on microtubular system in MCF-7 cell line. Methods Proliferation inhibition of MCF-7 cell line was evaluated by MTT assay. Cell cycle of MCF-7 cells was analyzed and the changes of a-tubulin protein and microtubule-associated protein 2 (MAP-2) protein were detected by flow cytometry. Results The IC50 of MCF-7 cells was 22.08 μg/mL. Solanine could induce MCF-7 cells arrested in S phaseand increase the levels of a-tubulin and MAP-2 in MCF-7 cell line. Conclusion Solanine could inhibit the MCF-7 cell proliferation by increasing a-tubulin and MAP-2 expression and inducing MCF-7 cells arrested in S phase.

Material basis of cytotoxicity in polysaccharide from Solanum nigrum / 中草药

Objective: To study the material basis of cytotoxicity in polysaccharide from Solanum nigrum. Methods: Crude products of polysaccharide were isolated from S. nigrum fruit with free protein being removed by Sevage method. The crude products were decolored by 10% H2O2 and precipitated by 95% ethanol treatment. Polysaccharide-protein complex was isolated by DEAE-52 fiber column and the relative molecular weight was detected by SDS-PAGE. MTT assay was used in detecting the cytotoxicity of polysaccharide-protein complex for MCF-7 cells in vitro. Then polysaccharide-protein complex was separated by SephadexG-200 gel column and the cytotoxicity was detected by MTT method. Results: Polysaccharide-protein complex was isolated by DEAE-52 fiber column and the relative molecular wight was detected by SDS-PAGE as 3.0×10 4 and 2.5×104 for the two polysaccharide-protein complexes, It suggested that IC50 was 804.51 μg/mL by MTT. Glycoproteins A andd B were gained by Sephadex G-200 gel column from polysaccharide-protein complex. The IC50 of glycoproteins A and B were 532.96 and 613.91 μg/mL, respectively. Conclusion: Material basis of cytotoxicity in polysaccharide from S. nigrum is two kinds of glycoproteins whose relative molecular weight are 3.0×104 and 2.5×104.

Inhibitory mechanism of oridonin on G2/M phase arrest in SGC-7901 cell growth / 中草药

Objective: To investigate the effect of oridonin on G2/M phase arrest of human stomach cancer SGC-7901 cell growth in vitro and its molecular mechanism. Methods: The inhibition of oridonin on SGC-7901 cells was detected by MTT assay. Effect of oridonin on the phase distribution of SGC-7901 cell cycle was observed by flow cytometry (FCM). Western blotting was used to analyze the expression of Cdk1 and CyclinB1. Results: IC50 of oridonin on SGC-7901 cells was 15. 64 μmol/L. The effect of oridonin on SGC-7901 cells presented the increasing percentages of cells in G2/M phase as the concentration of oridonin increased. With the increase of oridonin dose, the expression of Cdk1 and CyclinB1 proteins was remarkably decreased. Conclusion: Oridonin can down-regulate the expression of Cdk1 and CyclinB1, which may be the mechanism of arresting SGC-7901 cells in G2/M phase.

Studies of betuionic acid on cell cycle and related protein expressions on mice of bearing H22 tumor cells / 中国中药杂志

<p><b>OBJECTIVE</b>To study the betulonic acid on the cell cycle and related protein expressions on mice of bearing H22 tumor cells.</p><p><b>METHOD</b>Flow cytometray was used to observe the changes of betulonic acid on the cell cycle and P53 of H22 tumor cells. Immunohistochemistry was determined the espressions of PI3K and AKT.</p><p><b>RESULT</b>Increasing the doses of betulonic acid, the number of H22 cells in S phase and G2 phase was increasing gradually, it can speculate that when the betulonic acid act on cells, the cells were blocked in S and G2 phase and inhibited the protein expressions of PI3K and AKT.</p><p><b>CONCLUSION</b>Betulonic acid may be up-regulate the activity of P53 and inhibite the expressions of PI3K and AKT, so that it inhibited the survival pathway of tumor cells.</p>