Anticancer effect of total annonaceous acetogenins on hepatocarcinoma.

OBJECTIVE: To confirm the anticancer effect of total annonaceous acetogenins (TAAs) abstracted from Annona squamosa Linn. on human hepatocarcinoma. METHODS: The inhibitory effect of TAAs was demonstrated in H22-bearing mice. The potency of TAAs was confirmed as its 50% inhibiting concentration (IC50) on Bel-7402 cell under Sulfur Rhodamine B staining. Both underlying mechanisms were explored as cellular apoptosis and cell cycle under flow cytometry. Mitochondrial and recipient apoptotic pathways were differentiated as mitochondrial membrane potential under flow cytometry and caspases activities under fluorescence analysis. RESULTS: The inhibitory rate of TAAs in mice was 50.98% at 4 mg/kg dose. The IC50 of TAAs on Bel-7402 was 20.06 µg/mL (15.13-26.61µg/mL). Effective mechanisms of TAAs were confirmed as both of arresting cell cycle at G1 phase and inducing apoptosis dose- and time-dependently. Mitochondrial and recipient pathways involved in apoptotic actions of TAAs. CONCLUSION: TAAs is effective for hepatocarcinoma, via inhibiting proliferation and inducing apoptosis.

[The compounds from n-butanol fraction of Alpinia oxyphylla].

Nine compounds were isolated from the n-butanol fraction of 95% ethanol extract of the fruit of Alpinia oxyphylla Miq. with a combination of various chromatographic approaches, including MDS resin, silica gel, reverse phase C18 and preparative HPLC. On the basis of spectroscopic data analysis, they were elucidated as (1R, 4R, 10R)-1ß, 4α-dihydroxy-11, 12, 13-trinor-5, 6-eudesmen-7-one (1), 1ß, 4ß-dihydroxy-11, 12, 13-trinor-8, 9-eudesmen-7-one (2), oxyphyllenone A (3), oxyphyllenone B (4), rhamnocitrin (5), staphylionoside D (6), benzyl-1-O-ß-D-glucopyranoside (7), 2-O-ß-D-glucopyranosyl-(1S)-phenylethylene glycol (8), and (S)-1-phenylethyl-ß-D-glucopyranoside (9). Among them, compound 1 is a new sesquiterpene, named as oxyphyllenone C; compounds 8 and 9 are new natural products; compounds 2 and 6 were isolated from the genus Alpinia for the first time, and compound 7 was isolated from A. oxyphylla for the first time.

[Study on content of two sesquiterpenes in Alpinia oxyphylla].

OBJECTIVE: To develop an HPLC method for determining oxyphyllenodiol A (1) and teuhetenone A (2) contained in Alpinia oxyphylla and to compare the contents of the two components contained in medicinal materials and prepared herbal medicines in pieces sold in the market and different fractions. METHOD: HPLC and Waters sunfire C18 column (4. 6 mm x 250 mm, 5 microm) were adopted for gradient elute with the mobile phase of acetonitrile and water. The flow rate was 1.0 mL x min(-1) and the detection wavelength was 250 nm. RESULT: 1 and 2 showed a good linear relationship within the range of 0.1296 - 0.8640 microg and 0.1635 - 1.0900 microg respectively, with the average recoveries of 99.08% and 97.80%. Their content ranges were 0.0059% - 0.0149% and 0.0080% - 0.0164% in different samples. The mean value of 1 and 2 were 0.0085% and 0.0104% in the whole fruits, and 0.0137% and 0.0157% in the seeds. They were undetected in the nutshells. CONCLUSION: The method is so precise, accurate and highly reproducible that it can be used to determine the contents of oxyphyllenodiol A and teuhetenone A in A. oxyphylla. The contents of the two components are mainly extracted from the seeds, with certain difference among different samples. There are a higher contents and no significant difference in the salted and raw seeds.

Effects of L.F04, the active fraction of Lycopus lucidus, on erythrocytes rheological property.

OBJECTIVE: To study the effects of L.F04, the active fraction of Lycopus lucidus, on erythrocytes rheological property so as to investigate its mechanism in promoting blood circulation and removing blood stasis. METHOD: The effects of L.F04 (used for treatment for 10 days in different dosages) on deformability, aggregation and membrane liquidity of erythrocytes (MLE) as well as whole blood apparent viscosity (eta(b)) were examined on the basis of rat model of blood-stasis syndrome induced by venous injection of high molecular weight dextran. RESULT: As compared with the normal control group, the model group's RBC deformability and MLE were lower, and the aggregation of erythrocytes and eta(b) were higher. Compared with the model group, both L.F04 0.612 g/kg and 0.306 g/kg showed significant effect in improving deformability and inhibiting aggregation of red blood cells (RBC) and reducing blood viscosity. The trend of improving MLE was also shown. CONCLUSION: L.F04 could significantly improve the abnormal rheological property of erythrocytes.

[Research on the technology of soaking under pressure and cutting process of Radix Curcumae].

OBJECTIVE: To determine the best processing technology parameters. METHOD: The changes of the weights, volatile constituents, total extract quantities by MeOH or CHCl3 and curcumenol contents of the roots of Curcuma wenyujin and C. kwangsiensis roots before and after water soaking were determined, so as to determine the quality control index for the process of preparing Yujin slices. The shortest soaking and moistening time and the appropriate drying temperature were studied. RESULT AND CONCLUSION: (1) The weight was decreased and the quantities of volatile constituents were changed during water soaking process. But the total extracts quantities and the curcumenol contents didn't change obviously. So the quality control index was decided as soaking as short as possible. (2) The soaking time could be decreased by using high pressure. The soaking time was related with the short diameter of the cross section of the root. (3) The soaking under pressure technology is that soaking on -0.095 MPa for 0.5 h, then on 0.14 MPa for 10-16 h, and moistening for 36-48 h, cutting drying in room temperature or less than 40 degrees C.

[Chemical constituents from the seeds of Annona squamosa].

AIM: To study the antitumor active constituents of the seeds from Annona squamosa L. (Annonaceae). METHODS: Various chromatographic techniques were used to isolate and purify the constituents. Their physico-chemical properties and spectral data were determined to elucidate the structures. RESULTS: Eleven compounds were isolated and identified as annonaceous acetogenins: squamocenin (1), annotemoyin-2 (2), reticulatain-2 (3), squamocin-I (4), squamocin-B (5), squamocin (6), motrilin (7), squamostatin-D (8), squamostatin-E (9), cherimolin-1 (10), cherimolin-2 (11) from the ethyl alcohol extract of A. squamosa L. CONCLUSION: Squamocenin (1) is a new acetogenin. Annotemoyin-2 (2) and reticulatain-2 (3) were isolated from this plant for the first time.

Effects of active fractions from Lycopus lucidus L. F04 on platelet aggregation and thrombus formation.

OBJECTIVE: To study the effect of active fractions from Lycopus lucidus (L. F04) on platelet aggregation and thrombus formation so as to investigate its mechanism of promoting blood circulation and removing blood stasis. METHOD: The effects of Lycopus lucidus L. F04 on platelet aggregation induced by ADP in vivo, and thrombosis of artery-vein by-pass and thrombus formed in circuitous loop in vitro were examined using the blood stasis animal model of rats made by injecting high molecular weight dextran (HMWD). RESULT: The increase of maximum platelet aggregation rate induced by ADP in vivo in HMWD model was evidently inhibited with 0.408 g/kg or 0.204 g/kg or L. F04, and the effect showed a trend of concentration-dependence. As compared with the control, the thrombus weight in rat model of blood stasis increased clearly and its length showed only a trend of increase. Both 0.408 g/kg and 0.204 g/kg of L. F04 had the effect of resisting the thrombus formation, while 0.408 g/kg showed better effects of reducing the thrombus dry weight and wet weight. Both 0.408 g/kg and 0.204 g/kg of L. F04 could inhibit the thrombosis in artery-vein by-pass; the inhibition rates were 27.41% and 27.14% respectively. CONCLUSION: Lycopus lucidus L. F04 could significantly inhibit platelet aggregation and thrombus formation.

[Effects of active fractions from Lycopus lucidus L. F04 on erythrocyte rheology [correction of erythrocyterheology]].

Objective. To study the effects of active fractions from Lycopus lucidus (L. F04) on erythrocyte rheology so as to investigate its mechanism of promoting blood circulation and removing blood stasis. Method. The effects of L. F04 on erythrocytes deformability, aggregation, liquidity of membrane were examined in the present study and the blood stasis animal model of rats made by injecting high molecular weight dextran (HMWD) was used. Result. HMWD injection decreased red blood cells deformability (P<0.05) and liquidity of membrane (P<0.01) and increased erythrocytes aggregation (P<005). Compared with the model, both L. F04 0.612 g/kg and 0.306 g/kg improved red blood cells deformability (P<0.05), inhibited erythrocytes aggregation (P<0.05), and showed the trend of improving liquidity of membrane. But there was no concentration-dependence. Conclusion. L. F04 can significantly improve the deviant erythrocyte rheology characteristics.

[Two flavone glycosides from Chinese traditional medicine Amomum villosum].

OBJECTIVE: To study the water soluble constituents from Amomum villosum. METHOD: The constituents were separated and purified with chromatographic methods, identified by NMR, MS, UV and IR. RESULT: Two quercetin glycosides: quercitrin (quercetin-3-O-alpha-L-rhamnoside I) and isoquercitrin (quercetin-3-O-beta-D-glucoside II) were isolated and identified. CONCLUSION: I and II were isolated for the first time from A. villosum.

Murihexol, a linear acetogenin from Annona muricata.

Murihexol (1), a novel acetogenin, and three known ones, donhexocin (2), annonacin A and annonacin, have been isolated from the seeds of Annona muricata. Compound 1 is a C(35) acetogenin without any THF rings, with six hydroxyls in the long aliphatic chain. The hydroxyls of 1 and 2 are all located at C-4, C-10, C-15, C-16, C-19 and C-20, but their stereochemistries are different. The vicinal diol at C-15/C-16 in 1 is threo and C-19/C-20 has the erythro configuration; in 2 C-15/C-16 and C-19/C-20 all have the same threo configuration.