In vitro study on gastrointestinal absorption of FITC labeled pilose antler protein extraction / 药学学报

An in vitro detection method of the gastrointestinal absorption of Pilose Antler protein was established for mixed protein activity. Five bands of protein with molecular weight of 17.8-160 kD derived from the Pilose Antler were extracted and sufficiently labeled with FITC (FITC-PE). The stability and variation of FITC-PE in gastrointestinal circumstances were detected by native polyacrylamide gel electrophoresis and confocal laser scanning microscope. Results showed that the main component of FITC-PE kept invariant after being reacted with artificial gastric fluid and artificial intestinal fluid. The fluorescence signal was detected 20 min after administration in the valgus intestinal purse experiment, and three kinds of protein, with molecular weight of 45, 25, and 17.8 kD, were detected in the mixture of absorbent protein. The research laid the foundation for the further in vivo study of Pilose Antler protein. Meanwhile, it would be an in vitro screening method for the absorption, distribution and metabolism of mixed protein from traditional Chinese medicine.

Experimental study of velvet antler polypeptides against oxidative damage of osteoarthritis cartilage cells / 中国骨伤

<p><b>OBJECTIVE</b>To study reverse effect of the oxidative damage on cartilage cells of velvet antler polypeptides (VAPS), and to investigate the main mechanism of VAPS to protect cartilage cells through antioxidant.</p><p><b>METHODS</b>Fifteen Japanese white rabbits of 5-month-old were selected in this study. Animal model was established by method of Hulth osteoarthritis animal model. The anterior and posterior cruciate ligament and medial collateral ligament were cut off and medial meniscus were cut, articular cartilage cell cultured in vitro. Cells in the sham operation group was the normal control group, osteoarthritis cartilage cells in the model groups were added VAPS 6.25, 12.5, 25 microg/ml respectively. A group of animals were sacrificed every week form the ninth weeks(two months) and the cartilage cells were isolated and cultured. For 8 weeks,the reactive oxygen species level in chondrocytes were detected by DCFH-DA, the content of NO, SOD and GSH-Px in cell culture supernatant were detected by Griess method.</p><p><b>RESULTS</b>DCFH-DA detection of intracellular reactive oxygen species was (5.46 +/- 0.46)in the control group, (12.08 +/- 0.74) in the model groups. The model group compared with the control group by t test with the P value less than < 0.001. DCFH-DA detection of intracellular reactive oxygen species was (9.81 +/- 0.59)in VAPS 6.25 microg/ ml group, (7.83 +/- 0.63) in the VAPS 12.5 microg/ml group, (6.89 +/- 0.71) in the VAPS 25 microg/ml group, as compared with model group there were statistically significant difference (P < 0.05). The content of NaNO2, SOD and GSH-Px in osteoarthritis model group was (5.60 +/- 0.45) microM, (38.56 +/- 12.53) U/ml and (151.90 +/- 25.60) U, as compared with control group there were statistically significant difference (P < 0.001, P < 0.05); The content of NaNO2 was (4.34 +/- 0.39), M in VAPS 6.25 microg/ml group, (3.67 +/- 0.36) microM in the VAPS 12.5 microg/ml group, (3.20 +/- 0.27) microM in the VAPS 25 microg/ml group, as compared with model group there were statistically significant difference (P < 0.01). The content of SOD was (49.91 +/- 5.77) U/ml in VAPS 6.25 microg/ml group, (54.05 +/- 5.27) U/ml in the VAPS 12.5 microg/ml group, (57.44 +/- 5.70) U/ml in the VAPS 25 microg/mL group, as compared with model group there was statistically significant (P < 0.05). The content of GSH-Px was (172.50 +/- 18.65) U in VAPS 6.25 microg/ml group, (202.10 +/- 21.60) U in the VAPS 12.5 microg/ml group, (315.80 +/- 10.50) U in the VAPS 25 microg/ml group, the VAPS 12.5 microg/mL group and VAPS 25 microg/ml group was compared with model group, there were statistically significant difference (P < 0.01).</p><p><b>CONCLUSION</b>The VAPS have antioxidative damage effect of osteoarthritis cartilage cells within a certain range and dose-dependent manner. It may be the main mechanism for velvet antler polypeptides to treat osteoarthritis.</p>

Qualitative and quantitative determination of the main components of huanglianjiedu decoction by HPLC-UV/MS / 药学学报

<p><b>AIM</b>To establish a comprehensive HPLC analytical method of Huanglianjiedu decoction.</p><p><b>METHODS</b>This study was performed by HPLC-UV/MS to identify the chemical constituents of the whole and individual herbs of the "Huanglianjiedu decoction". Zorbax Extend C18 (150 mm x 4. 6 mm ID, 5 microm) column was used; the mobile phase was composed of acetonitrile (A) and water (B, with 0.5% acetic acid) with gradient elution; the flow rate was 1.0 mL x min(-1) and the column temperature was setup at 25 degrees C. The detection wavelength was 254 nm.</p><p><b>RESULTS</b>The chromatogram of Huanglianjiedu decoction showed 21 main peaks. Peaks 1, 2, 5 and 18 were from Gardenia jasminoides Ellis, Peaks 8, 13, 14, 15, 16, 17, 19 and 21 from Scutellaria baicalensis Georgi. While 10 from Coptis chinensis Franch and 20 from Phellodendron amurense Rupr., Peaks 3, 4, 6, 9, 11 and 12 came from them together. Peak 7 presented in the chromatograms of the herbs except Gardenia jasminoides Ellis. By comparison of the retention time, the on-line UV spectra and MS spectra, 11 peaks were identified as 5 (geniposide), 9 (jatrorrhizine), 10 (coptisine), 11 (palmatine), 12 (berberine), 13 (baicalin), 15 (oroxin A), 17 (wogonoside), 19 (baicalein), 20 (obaculactone), 21 (wogonin), then eight of them were quantified by HPLC-UV.</p><p><b>CONCLUSION</b>The method could represent the characteristics of Huanglianjiedu decoction, and it could be used to evaluate the quality and quantity of Huanglianjiedu decoction. It distinguished between Coptis chinensis Franch and Phellodendron amurense Rupr. by HPLC for the first time.</p>

Study on antitumor activities of huanglian jiedu decoction / 中国中药杂志

<p><b>OBJECTIVE</b>To study the antitumor activity of Huanglian Jiedu decoction (HLJDT).</p><p><b>METHOD</b>Antitumor activities were tested in mice with experimental tumor H22 in vivo, and the thymus index, spleen index and tumor inhibitory rate were evaluated. The effects on cancer cells from human were investigated in vitro using serum pharmacological approach. Swille, SPC-A-1, SGC-7901 and MCF-7 cancer cells were incubated in culture media containing serum from mice medicated with HLJDT. The inhibitory effects of HLJDT serum were observed by MTT assay.</p><p><b>RESULT</b>HLJDT showed significant antitumor activities on H22 in mice. All of the HLJDT serum in different dosage groups could highly inhibit the proliferation of 4 cancer cell lines from human.</p><p><b>CONCLUSION</b>The HLJDT can significantly inhibit the tumor H22 in mice in a dose-dependent manner, the drug serum has obvious anticancer effects against Swille, SPC-A-1, SGC-7901 and MCF-7.</p>

Comparison between antitumor effect and chemical constituents of Huanglian Jiedu decoction and that of serum containing Huanglian Jiedu decoction / 中国中药杂志

<p><b>OBJECTIVE</b>To make a comparison between the antitumor effect and the chemical constituents of Huanglian Jiedu decoction (HLJDT) and that of serum containing HLJDT.</p><p><b>METHOD</b>Based on the established chromatographic fingerprint of HLJDT, analysis and comparison were made between the HPLC fingerprints of rat serum samples obtained after orally taking HLJDT and those of control rat serum samples. The different effects on NCI-H446 and Bel-74024 cancer cells from human were investigated in vitro using HLJDT and its serum. The inhibitory effects of HLJDT and its serum were observed by MTT assay.</p><p><b>RESULT</b>Ten compounds of HLJDT and some metabolites were detected after oral administration of HLJDT, and however some main compounds of HLJDT were not detected in serum. Both HLJDT and its serum in different dosage groups could inhibit the proliferation of NCI-H446 and Bel-7402 cancer cells from human in a dose-dependent manner, but inhibitory grade was different in the two cancer cell lines. HLJDT had more inhibitory effect on Bel-7402 than on NCI-H446, on the other hand serum containing HLJDT had the same inhibitory effect on Bel-7402 and NCI-H446.</p><p><b>CONCLUSION</b>The reason for inhibitory grade change was that the proportion of concentration of many compounds in serum containing HLJDT was different to that in HLJDT, which should be subject to thorough investigation so as to illuminate the pharmacology and active mechanism of HLJDT.</p>

Comparative study on anti-hypercholesterolemia activity of diosgenin and total saponin of Dioscorea panthaica / 中国中药杂志

<p><b>OBJECTIVE</b>To compare the anti-hypercholesterolemic and cholesterol absorption inhibitory activities between total saponin of Dioscorea panthaica (TSDP) and diosgenin (Dio).</p><p><b>METHOD</b>TSDP and Dio were given ig or i.p. to mice or rats treated with cholesterol feed to evaluate their preventive and therapeutic effect on hypercholesterolemia. TSDP or Dio and cholesterol were mixed with pig bile to form the micelle, then the freeing cholesterol was detected to evaluate inhibitory effect of the both compounds on cholesterol absorption.</p><p><b>RESULT</b>Dio (80 and 160 mg.kg-1) showed significantly therapeutic and preventive effect on hypercholesterolemia in mice, while TSDP showed a certain preventive activity only at a big dose (400 mg.kg-1). The intraperitoneal injection of Dio (20 and 40 mg.kg-1) to mice suffered from hypercholesterolemia was effective, but TSDP showed no effective. The serum total cholesterol level was decreased when rats were pre-treated with TSDP (200 and 400 mg.kg-1, ig) and Dio (200 and 100 mg.kg-1, ig). However, the hypercholesterolemia-preventing activity of Dio was stronger than that of TSDP. In addition, inhibitory effect of Dio on cholesterol micelle formation was still stronger than that of TSDP.</p><p><b>CONCLUSION</b>The preventive and therapeutic activity of Dio against hypercholesterolemia indused by cholesterol in mice or rats is stronger than that of TSDP. The anti-hypercholesterolemia mechanism of Dio is probably related with its cholesterol absorption inhibitory activity.</p>

Intestinal bacteria metabolism of TSDP and characterization of metabolites in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To study the rat intestinal bacteria metabolism of total saponins of Dioscorea pathaica (TSDP) in vitro, and characterize the metabolites in serum and urine of rats after oral administration of TSDP 900 mg.kg-1.</p><p><b>METHOD</b>TSDP metabolites were detected with thin-layer chromatography (TLC) and combination of electrospray ionization mass spectrometry (ESI-MS) and sequential tandem mass spectrometry (MSn).</p><p><b>RESULT</b>In vitro, TSDP was decomposed easily by rat intestinal bacteria, and metabolites DP-1, DP-2, DP-4, DP-5 and diosgenin (Dio) were observed with prolongation of incubation time by ESI-MS2. In vivo, in the full-scan positive mass spectrum of the rat urine sample, the ion peak at m/z 415 (M-H) and its characteristic fragmentations at m/z 397 and m/z 271 in the MS/MS spectrum were identified with that of metabolite Dio, therefore metabolite Dio was deduced to exist in the rat urine, and metablite Dio was allso detected in the rat serum sample.</p><p><b>CONCLUSION</b>TSDP is decomposed easily by rat intestinal bacteria and metabolite diosgenin is absorbed into blood after oral administration of TSDP.</p>