Fraction n-Butanol of Radix Notoginseng Protects PC12 Cells from Aß25-35-Induced Cytotoxicity and Alleviates Cognitive Deficits in SAMP8 Mice by Attenuating Oxidative Stress and Aß Accumulation.

Chinese medicine has been used for Alzheimer's disease (AD) treatment for thousands of years with more effective and fewer side effects. Therefore, developing effective potential candidates from Chinese medicine against AD would be considered as critical and efficient therapy for AD treatment. This study was designed to evaluate the neuronal protective effect of fraction n-butanol (NB) of Radix Notoginseng on Aß25-35-induced PC12 cells, explore the effect of the tested fraction on spatial learning and memory, and characterize the impacts of fraction NB on antioxidant enzymes, Aß production, and APP and BACE1 expressions. The results revealed that fraction NB could promote proliferation of PC12 cells and protect and rescue PC12 cells from Aß25-35-induced cell death. Moreover, fraction NB could improve spatial learning and memory impairments of senescence-accelerated prone8 (SAMP8) mice and attenuate oxidative stress and reduce the production of Aß by inhibiting the expressions of APP and BACE1 in the brains of SAMP8 mice. The result of single dose acute toxicity assay showed that fraction NB had a mild toxicity in vivo. The pronounced actions against AD and in vivo low toxicity of fraction NB suggest that fraction NB may be a useful alternative to the current AD treatment.

[Preliminary establishment of integration of Alzheimer's disease and blood stasis syndrome tree shrew model and evaluation of intervention of Panax notoginseng saponins].

To establish the integration of Alzheimer's disease(AD) and blood stasis syndrome tree shrew model. Panax notoginseng saponins (PNS) was used to intervene the model to testify the stability of the model. The level of blood stasis of each group in the tree shrew model was evaluated by analyzing five traditional Chinese medicine(TCM) characterizations, four blood coagulation indexes, plasma nitric oxide (NO) level, plasma superoxide dismutase (SOD) level in each group. Hematoxylin and eosin(HE) staining was used to observe the morphological changes of brain hippocampal neuron cell of each group. Immunohistochemical staining was used to assay the ChAT and SYP levels in brain hippocampus of each group.The blood stasis characterization of the integration of disease and syndrome group was more obvious than the AD group, and that of the drug administration group was lower than that of the integration of disease and syndrome group. Aß1-42, APP, P-Tau, ChAT and SYP level of AD group were lower than those in the blank group, which were further reduced in the model of integration of disease and syndrome. However, the administration of PNS relieved the reduction, indicating that the AD and blood stasis integration syndrome tree shrew model is stable.

Cestrum nocturnum Flower Extracts Attenuate Proliferation and Induce Apoptosis in Malignant Cells through Inducing DNA Damage and Inhibiting Topoisomerase II Activity.

Most of the existing chemotherapeutic drugs have plenty of side effects. Chinese herbal medicine has been used for pharmaceutical and dietary therapy for thousands of years with more effective and fewer side effects. Cestrum nocturnum (CN) has long been used to treat digestive diseases for centuries in China. Our previous study first proved that the n-butanol part isolated from the flowers of CN produced an inhibitory effect on the proliferation of malignant cells. However, the fractions responsible for the antiproliferation effect of n-butanol part from CN flowers and related mechanisms remain unknown. Thus, in this study, we extracted fractions C4 and C5 from n-butanol part of CN flowers and investigated their immune toxicity and antitumor activities. It was found that fractions C4 and C5 exhibited great cytotoxicity to cancer cell lines but had low immune toxicity towards T and B lymphocytes in vitro. The tested fractions also attenuated proliferation and induced apoptosis at G0/G1 and G2/M phases in Bel-7404 cells through inducing DNA damage and inhibiting topoisomerase II relaxation activity. These results suggest that fractions C4 and C5 may represent important sources of potential antitumor agents due to their pronounced antitumor effects and low immune toxicity.

Neuroprotective Properties of Panax notoginseng Saponins via Preventing Oxidative Stress Injury in SAMP8 Mice.

Inhibiting oxidative damage in early stage of Alzheimer's disease (AD) is considered as a strategy for AD treatment. Our previous study has shown that Panax notoginseng saponins (PNS) have an antiaging action by increasing the levels of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-PX) in the serum of aged rats. In this study, we aimed to investigate the effects of PNS on antioxidant enzymes and uncoupling proteins (UCPs) involved in oxidative stress in AD mice. The results showed that PNS prevented neuronal loss in hippocampal CA1 region and alleviated pathomorphological change of neurons in CA1 region. Moreover, PNS inhibited the production of 8-hydroxydeoxyguanosine (8-OHdG), enhanced the expressions and activities of SOD, CAT, and GSH-PX, and improved the mRNA and protein levels of UCP4 and UCP5 in the brains of SAMP8 mice. Together, our study shows that PNS has the ability to protect neurons in AD brain from oxidative stress damage through attenuating the production of 8-OHdG, enhancing the activities of antioxidant enzymes and the expressions levels of UCP4 and UCP5. Accordingly, PNS may be a promising agent for AD treatment.

[Study on Chemical Composition of Phyllanthus emblica].

OBJECTIVE: To study the chemical constituents of Phyllanthus emblica. METHODS: The chemical constituents were isolated and purified by silica gel, polyamide and Sephadex LH-20 chromatography. Their structures were elucidated by physicochemical proper- ties and spectral analysis. RESULTS: 13 compounds were isolated and identified as Triacontanol (1), Triacontanoic acid (2), ß-Amyrin ke- tone (3), Betulonic acid (4), Daucosterol (5), Lupeol acetate (6), ß-Amyrin-3-palmitate (7), Gallic acid (8), Betulinic acid (9), Ursolic acid (10), Oleanolic acid (11), Quercetin (12) and Rutin (13). CONCLUSION: Compounds 1,2,4,6,7,9,10 and 11 are obtained from Phyllanthus emblica for the first time.

[Protective effect of serum containing n-butanol fraction from Gynostemma pentaphyllum on NG108-15 cell apoptosis induced by Aß25.35 protein].

OBJECTIVE: To investigate the inhibition effect of serum containing n-butanol fractions from Gynostemma pentaphyllum on NG108-15 cell apoptosis induced by Aß25-35 protein in vitro. METHODS: The apoptosis of NG108-15 cells induced by Aß25-35 protein in vitro was evaluated by immunofluorescence and flow cytometry assay. The cellular Caspase-3 level during the apoptosis was determined by ELISA. RESULTS: The serum containing n-butanol fractions from Gynostemma pentaphyllum significantly inhibited the NG108-15 cells apoptosis induced by Aß25-35 protein in vitro,and decreased the cellular Caspase-3 level. CONCLUSION: The inhibition effect of n-butanol fractions from Gynostemma pentaphyllum on NG108-15 cell apoptosis induced by Aß25.35 protein is likely related to its potency on reducing of cellular Caspase-3 level.

[Study on the effect of extracts from the leaves of Phyllanthus emblica on immune function of mice].

OBJECTIVE: To explore the effect of extracts from the leaves of Phyllanthus emblica (PLFs) on the immune function of mice. METHODS: 70 Kunming mice were choosed to conduct the acute toxicity test of PLFs. The mice were randomly divided into four groups: PLFs high-dosage group, mid-dosage group, low-dosage group and control group. The high,mid,low-dosage groups were treated with PLFs 1.982, 0.991 and 0.496 g/kg respectively per day. The same volume of double distilled water was given to the control group. All by intragastric administration for 7 d. The animals were killed and indexes of thymus and spleen were calculated. The expurgation index K and phagocyte index a were detected after the mice being injected with a dilute India ink through caudal vein. In addition, prepared spleen cells conventionally,the activity of Natural Killer cells was measured and the proliferation of T and B cells were detected. The effect of the extracts on serum hemolysin was detected after the SRBC was injected into the enterocoelia. RESULTS: The LD50 of PLFs was 9. 911 g/kg. Compared with the control group, the indexes of thymus and spleen in the treatment groups had no markedly difference (P > 0.05). The high- and mid-dosage groups could obviously improve the expurgation index K (P < 0.05), phagocyte index alpha (P < 0.05) and NK cell activity (P < 0.05). CONCLUSION: The extracts from Phyllanthus emblica leaves can promote nonspecific immunity immune function in mice.

[Effect of PNS on the activity and content of BACE1 in the brain of SAMP8 mice with Alzheimer's disease].

OBJECTIVE: To explore the effect of Panax notoginseng saponin (PNS) on the activity and content of beta-secretase in the brain of senescence accelerated mouse-prone 8 (SAMP8) mice with Alzheimer's disease. METHODS: Totally 32 SAMP8 mice were randomly divided into the normal control group, the high dose PNS group (200 mg/kg), the low dose group (100 mg/kg), and the huperzine A group (0.3 mg/kg), 8 in each group. Equal volume of double distilled water was given to those in the normal control group. All medication was given by gastrogavage, once daily for two successive months. The activity of BACE1 was assayed by direct immunofluorescent method (DIF). The content of BACE1 protein was detected by Western blot. RESULTS: The relative fluorescence units (RFU/microg) was 2.008 +/- 0.031 in the high dose PNS group, 2.221 +/- 0.029 in the low dose PNS group, and 2.267 +/- 0.076 in the huperzine A group, all lower than that in the normal control group (2.403 +/- 0.058; all P < 0.01). The content of BACE1 protein was 0.900 +/- 0.028 in the high dose PNS group, 1.000 +/- 0.032 in the low dose PNS group, and 0.837 +/- 0.080 in the huperzine A group, all lower than that in the normal control group (2.210 +/- 0.074, all P < 0.01). CONCLUSION: PNS higher than 100 mg/kg could decrease the activity of BACE1 and down-regulate the content of BACE1 protein in the brain of SAMP8 mice.

[Effect of Panax notoginseng saponins on APP gene transcription in the brain tissue of SAMP8].

OBJECTIVE: To study the effect of Panax notoginseng saponins (PNS) on learning and memory ability and APP gene transcription in the brain tissue in senescence accelerated mouse prone 8 (SAMP8). METHODS: SAMP8 were randomly divided into high-does PNS group, low-does PNS group, huperzin A group and model group,the treatment groups were treated with the designed drugs respectively by intragastric administration for 4 consecutive weeks. The same volume of double distilled water was given to model group. After treatment, the abilities of learning and memory of the mice were tested with morris water maze, the mRNA content of APP was assayed by reverse transcription (RT) and real-time polymerase chain reaction (RT-PCR). RESULTS: PNS could improve the abilities of learning and memory, high-does PNS could reduce the mRNA content of APP in the brain tissue of SAMP8. CONCLUSION: PNS can improve the abilities of learning and memory of SAMP8, the mechanism may be relevant to down-regulating the expression of APP gene at transcriptional level.

Progallin A isolated from the acetic ether part of the leaves of Phyllanthus emblica L. induces apoptosis of human hepatocellular carcinoma BEL-7404 cells by up-regulation of Bax expression and down-regulation of Bcl-2 expression.

AIM OF THE STUDY: This study was aimed to investigate the effects of Progallin A isolated from the acetic ether part of the leaves of Phyllanthus emblica L. on apoptosis in human hepatocellular carcinoma BEL-7404 cells and its possible mechanisms, and to measure the immune toxicity of Progallin A in vitro. MATERIALS AND METHODS: Progallin A was isolated from the acetic ether part of the leaves of Phyllanthus emblica L. with column chromatography. The proliferation of spleen lymphocytes and the viability of BEL-7404 cells were assessed with MTT assay. Inverted microscope, light microscope and fluorescence microscope were utilized to observe the morphological changes of BEL-7404 cells respectively. AnnexinV/PI double labeling and TUNEL assay were used to detect early apoptosis and DNA fragmentations of BEL-7404 cells respectively. In addition, cell cycle distribution was analyzed by using flow cytometry (FCM). Bax and Bcl-2 protein levels were determined by immunofluorescence staining and western blot respectively. RESULTS: The results showed that Progallin A had low immune toxicity and the proliferation of BEL-7404 cells was strongly inhibited by Progallin A in a time- and dose-dependent manner and that the characteristics of apoptosis of BEL-7404 cells were observed. The results also showed that apoptosis rates and the number of apoptotic cells significantly increased with prolongation of the action time. The results of flow cytometry (FCM) indicated that Progallin A induced significant G1/M and G2/M arrest of BEL-7404 cells. Immunofluorescence staining and western blot showed that the expression of Bax was found to be noticeably up-regulated and the expression of Bcl-2 was down-regulated significantly. CONCLUSIONS: These results indicate that Progallin A has low immune toxicity in vitro and induces apoptosis of human hepatocellular carcinoma BEL-7404 cells which is related to the G1/M and G2/M arrest, and it exerts its apoptotic effect by up-regulation of Bax expression and down-regulation of Bcl-2 expression.

[The effect of gallic acid extracted from leaves of Phyllanthus emblica on apoptosis of human hepatocellular carcinoma BEL-7404 cells].

OBJECTIVE: To explore the effect of gallic acid extracted from Leaves of Phyllanthus emblica on the apoptosis of BEL-7404 cells. METHODS: MTT assay was applied to detect the influence on prolifetation in vitro. Inverted microscope was utilized to observe the morphological changes after BEL-7404 cells were treated with gallic acid. Annexin V/PI double label method was used to detect earlier period apoptosis cells and Tunel was applied to calculate the apoptosis rates. RESULTS: Gallic acid could restrain the BEL-7404 cells proliferation at diffierent levels in a time and concentration dependent manner. The typical morphological changes of apoptosis were observed after BEL-7404 cells were treated with gallic acid. Annexin V/PI double label method and Tunel method showed that the viable apoptotic cell and apoptosis rates added as action time prolonged. CONCLUSION: Gallic acid can restrain the BEL-7404 cells proliferation and induce apoptosis, and its effect on apoptosis is time dependent.

[Analysis of essential oil from the leaves and fruits of Eucalyptus tereticornis in Guangxi province by GC-MS].

OBJECTIVE: To Analyze the chemical composition and their relative contents of essential oil from the leaves and fruits of Eucalyptus tereticornis in Guangxi province. METHODS: The column temperature was controlled by a program with a capillary column HP-5 MS, and the MS analysis was performed with EI and quadrupole mass analyzer. The chemical compositions were identified by NIST98 searching and mass spectra comparing, and their relative contents were determined by using normalization method of chromatographic peak areas. RESULTS: 39 compounds constituting 96.69% of the oil of leaves were identified. The major components were eucalyptol (27.93%), 1R-alpha-pinene (22.60%), isopinocarveol (8.71%); 36 compounds constituting 93.50% of the oil of fruits were identified. The major components are 1R-alpha-pinene (32.88%), eucalyptol (13.64%), D-limonene (8.31%). CONCLUSION: The GC-MS is a simple, rapid and sensitive method.

[Inhibitive effects of Panax notoginseng saponins on expression of Abeta(1-40), Abeta(1-42) protein in SAMP8's brain].

OBJECTIVE: To observe inhibitive effects of Panax notoginseng saponins on expression of Abeta(1-40), Abeta(1-42) protein in SAMP8's brain. METHODS: Amount of Abeta(1-40), Abeta(1-42 immuno-positive neurons was detected in parietal cortex and hippocamp in their brains under high power lens (40 x) by immunohistochemistry analysis. RESULTS: PNS could reduce the amount of Abeta(1-40), Abeta(1-42) protein in parietal cortex and hippocamp. CONCLUSION: PNS can reduce the amount of Abeta(1-40), Abeta(1-42) protein in SAMP8's brain.

[Anti-tumor mechanism of active components from extract of Actinidia rufa root].

OBJECTIVE: To observe effect and mechanism of n-Butanol lysate of alcohol extracts from Actinidia rufa root (monomer of R6,R8). METHOD: Tunel, Wright's stain with Giemsa's stain dyeing, and Hoechst 33258-PI double dyeing assay were used to detect the apoptosis of SGC7901 tumor cells treated with R6, R8. The SGC7901 tumor cells were randomly divided into control group and two treatment groups administered 0.05 g x L(-1) R6, R8, respectively, for 72 h). FCM assay was used to detect the apoptosis. Agarose electrophoresis assay was used to detect DNA strand break of tumor cells and reveal anti-tumor action mechanism. RESULT: The apoptosis percentage of the tumor cell in 24 h, 48 h, 72 h was (17.08 +/- 2.78)% , (29.68 +/- 2.96)%, (52.46 +/- 3.81)%; (14.75 +/- 2.14)%, (27.35 +/- 3.79)%, (45.64 +/- 5.24)%, respectively, for the treatment group, significantly higher than that in the control group (1.94 +/- 1.55)%, (2.78 +/- 1.84)%, (11.8 +/- 2.79)% (P < 0.01) by tunnel assay. Wright's stain with Giemsa's stain dyeing assay, Hoechst 33258-PI and FCM double dyeing assay showed same action. R6 and R8 had the effect of inducing the DNA histogram of tumor cells (P < 0.01). CONCLUSION: The anti-tumor mechanisms may be associated with inducing the injury of DNA and stimulating apoptosis.

[Analysis of the volatile oils chemical constituents of roots of Actinidia deliciosa].

OBJECTIVE: To analyze the volatile oils chemical constituents of roots of Actinidia deliciosa. METHODS: The volatile oils fraction of roots of Actinidia deliciosa. were extracted by water vapor distilling, and then the constituents were separated and identified, by GC-MS. RESULTS: 16 compounds were identified, accounting for 89.37% of all quantity. CONCLUSION: The principal volatile oils chemical constituents are Phenol, 2,4-bis(1,1-dimethylethl)-; 2-Propenoic acid, 3-(4-methox yphenyl)-, ethyl ester; 9-Octadecenoic acid (Z)-, methyl ester; Cyclotetrasiloxane, octamethyl-.

[Experimental study on antitumor effect of extracts from Cestrum nocturnum in vivo].

OBJECTIVE: To observe the antitumor effects of extracts from Cestrum nocturnum (CN) in vivo. METHODS: The S180-mice model was used to observe the tumor-inhibition rate of CN and the H22-mice model was used to test the survival time. RESULTS: The experiment in S180-mice demonstrated that the n-butanol and polysaccharides extracts from CN had obvious effects on tumor inhibition. Its inhibitory rates were 52.30%, 46.75%, 42.28%, 43.19%, 37.96% and 31.82% respectively in the mice administrated the n-butanol and polysaccharides extracts from CN with 30 mg/kg, 20 mg/kg and 15 mg/kg weight dosage. It was noted that tumor formation postponed in mice treated with the n-butanol and polysaccharides extracts from CN compared with the control panel and tumor growth became slower; The n-butanol and polysaccharides extracts from CN could also greatly enhance the life span of mice with H22 ascitic tumors by 116.43%, 44.52%, 20.54%, 109.52%, 112.61% and 115.01%, respectively. The inhibit effects of n-butanol fraction extracts from CN had direct relationship with dose, while the polysaccharides fraction extracts from CN had no obvious relationship with dose. CONCLUSION: The n-butanol and polysaccharides extracts of CN are able to inhibit tumor growth and prolong the lifetime of the tumor-bearing mice in a dose-dependent pattern.

[Studies on the chemical constituents of the ethyl acetate portion of Nervilia fordii].

OBJECTIVE: To study the chemical constituents of the ethyl acetate portion in the herb of Nervilia fordii from guangxi. METHODS: The constituents were separated and purified by using column chromatography with silica gel. These compounds were identified by their physical and spectral data. RESULTS: Five compounds were isolated and identified as norleucine (crystal I), 24 (S/beta)-dihydrocycloeucalenol-(E)-p-hydroxy cinnamate (crystal II) , rhamnocitrin (crystal III), rhamnazin (crystal IV), daucosterol (crystal V). CONCLUSION: Compounds I , II, III, IV, V were isolated from this plant for the first time.

[Study on the anticancer effects of extracts from roots of Livistona chinensis in vitro].

OBJECTIVE: To observe the effects of extracts from root of Livistona chinensis on the growth inhibition in seven carcinoma cell lines. METHODS: The growth inhibition was analyzed by MTT, cell colony and cell growth curve measuremen technique in the stomach carcinoma SGC7901, Lymphocytic leukemia L1210, Lymphoid neoplasm P388D1, tumor of cervix uteri Hela, hepar carcinoma hele 7404, melanoma B16 and mouse neuroblastomax rat glioma hybrid NG108-15 cells lines. RESULTS: The growth of all tumor cells were inhibited by ethyl acetate of alcohol extract from roots of Livistona chinensis. The growth of all tumor cells were not affected by low dose extracts (0.5 microg/ml). The growth of all tumor cells were obviously inhibited by higher dose extracts (5.0 microg/ml). The growth of all tumor cells were inhibited in growth curve measurement. CONCLUSION: The results show that ethyl acetate of alcohol extracts from roots Livistona chinensis possesses the role of antitumor in cell culture.

[Study on anti-tumor effect of extractions from roots of Actinidia deliciosa].

OBJECTIVE: To study the anti-tumor effect of ADEE and ADBE in mice. METHODS: The models of S180 entity tumor mice and H22 bearing tumor mice were established to observe the effect of ADEE and ADBE on inhibiting S180 entity tumor growth and prolonging life time of H22-mice, and the effect on spleen and thymus index to S180 entity tumor. RESULTS: Both ADEE and ADBE had ohvious anti-tumor effects to S180-mice (P < 0.05, P < 0.01) and could prolong H22-mice life (P < 0.05, P < 0.01). In certain degree, ADEE and ADBE could improve the immune function of bearing tumor mice. CONCLUSION: ADEE and ADBE have obvious anti-tumor effect.

[Study on anticancer effect in vivo of active fraction from Nervilia fordii].

OBJECTIVE: To determine the active fraction with anticancer effect in vivo from Nervilia fordii. METHODS: The effective petroleum ether extract and ethyl acetate extract parts preliminary were selected in vitro, then anticancer experiments in vivo were done by S180-mice and H22-mice models. RESULTS: Petroleum ether extract and ethyl acetate extract parts both had obvious anticancer effects to S180-mice and H22-mice, and could prolong H22-mice life. Meanwhile, they could improve the immunoloregulation of mice. CONCLUSION: It is the first time that the petroleum ether extract and ethyl acetate extract of Nervilia foadii are proved to be the effective anticancer fractions in vivo. On this basis, the further studies are needed on active principles or principle group with anticancer effect and the characteristics of this effect in Nervilia foadii.