[Influence of different preparation on the content of carbohydrate and physicochemical properties of polysaccharides from raw paeoniae radix alba and stir-baked paeoniae radix alba].

OBJECTIVE: To observe the difference of two preparation methods on the content of carbohydrate and physicochemical properties of polysaccharides from raw Paeoniae Radix Alba and stir-baked Paeoniae Radix Alba. METHODS: Polysaccharides extracted from raw Paeoniae Radix Alba and stir-baked Paeoniae Radix Alba with water were precipitated by ethanol and named as BSEP-S and BSEP-C, respectively. In the same way, those deposited by acetone were named as BSAP-S and BSAP-C. Their physicochemical properties, including extraction yield, the content of carbohydrate, elemental composition and monosaccharide composition, were determined. RESULTS: Extraction yield, sugar content and uronic acid content of BSEP-S was 1.56%, 80.56% and 3.33% , BSEP-C 1.18%, 80.79% and 5.47%, BSAP-S 1.58%, 86.50% and 3.79%, and BSAP-C 1.54%, 81.64% and 3.25%, respectively. In addition, monosaccharide compositions showed that glucose was the main monosaccharide and successively reached 92.21%, 87.30%, 91.45% and 86.62%. CONCLUSION: Yield and content of carbohydrate from raw Paeoniae Radix Alba and stir-baked Paeoniae Radix Alba by water extraction-ethanol precipitation are a little higher than that by water extraction-acetone precipitation, but monosaccharide compositions are almost the same. Different preparation has significant impact on the yield and the content of carbohydrate in Paeoniae Radix Alba by stir-baked method, and it can decrease the dissolution of polysaccharide.

[Anti-HIV activities of Achyranthes bidentata polysaccharide sulfate in vitro and in vivo].

Achyranthes bidentata polysaccharide sulfate (ABPS) was a sulfated derivate derived from Achyranthes bidentata polysaccharide (ABP) which was isolated and identified from Chinese herb Achyranthes bidentata. The anti human immunodeficiency virus type 1 (HIV-1) activities were studied in vitro and in vivo. ABPS was found to inhibit HIV-1 reverse transcriptase and integrase with the 50% inhibiting concentration (IC60) of (2.948 +/- 0.556) micromol x L(-1) and (0.155 +/- 0.030) micromol x L(-1), respectively, but the parent compound ABP was not effective. ABPS inhibited HIV-1 P24 antigen with IC50 of (0.082 +/- 0.044) micromol x L(-1) and selective index (SI) of > (358 +/- 148) in MT-4 cell cultures acutely infected with HIV-1 IIIB virus, and with IC50 of (11.80 +/- 5.90) micromol x L(-1) and SI of > (24.2 +/- 12.1) in PBMC cell cultures acutely infected with clinical isolated zidovudine resistant HIV-1 virus, but there was no activity even at its concentration of 500 micromol x L(-1) in latent infection of H9/HIV-1 IIIB cell cultures. 5% sera taken from rats after intraperitoneal injection from rats with ABPS 125 mg x kg(-1) once or mice with 3 mg x kg(-1) qd for 20 days effectively inhibited HIV-1 P24 in MT-4 cell cultures, but those had no inhibitory effect when given orally. The results suggested that ABPS is a promising HIV-1 inhibitor, active on HIV-1 reverse transcriptase, integrase in vitro and HIV-1 P24 antigens in cell cultures, it was well absorbed by intraperitoneal injection but poor in oral bioavailability. It warrants further study.

[Targeting glycyrrhetinic acid to hepatic stellate cells in treating rat liver fibrosis].

OBJECTIVES: We synthesized M6P26-HSA as a carrier for hepatic stellate cells (HSC) and coupled it with glycyrrhetinic acid (GA) to get a new conjugate GA-HSA-M6P26. Its organ distribution, specific combination with HSC and anti-fibrotic effect on livers were studied. METHODS: The GA-HSA-M6P26 was labeled with 125I and its organ distribution was detected radiologically. Selective combination of GA-HSA-M6P26 was observed with double immunocytochemic staining and collagen staining of the liver preparations was carried out using Sirius red staining method. The effect of the conjugate on mRNA expression of type I procollagen was studied with real-time PCR in vivo. RT-PCR was used for the effect on mRNA expression of alpha-SMA, MMP-9 and TIMP-1. RESULTS: 10 minutes after GA-HSA-M6P26 i.v. injection, 55%+/-5% of it was distributed in the livers. Double immunocytochemic staining showed that most of GA-HSA-M6P26 was taken up by HSC. With GA- HSA- M6P26 treatment, the collagen deposition in the liver decreased significantly compared with GA and M6P26-HSA treated rats. Similarly, the mRNA expression of type I procollagen and alpha-SMA dropped significantly. As to MMP-9 and TIMP-1, no significant change was shown. CONCLUSION: GA-HSA-M6P26 was selectively delivered to HSC and it showed a significant anti-fibrotic effect on rat liver fibrosis.

[Effects of LbGp on the intracellular free calcium concentration of cardiomyocytes induced by hypoxia and KCl].

OBJECTIVE: Hypoxia/KCl injury model in the cultured neonatal rat cardiomyocytes (CMs) was established to investigate the protective effect of Lycium barbanun Glycopeptide (LbGp) on calcium overload. METHOD: Cultured neonatal rat CMs were divided into three groups, namely normal control, hypoxia groups and LbGp-treated group. CMs in LbGp-treated group and hypxia group were cultured in an incubator ventilated with 95% N2 and 5% CO2 with or without LbGP. CMs viability under hypoxia was measured by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide colorimetry (MTT). The intracellular free calcium concentration in cardiomyocytes was measured by laser confocal microscope with Fura-3/AM as a calcium indicator. The protective effects of LbGp on the CMs treated by KCl (60 mmol x L(-1)) was observed. RESULT: As compared with normal controls, the degree of MTT metabolism was significantly reduced (P < 0.01) in hypoxic group and slightly reduced in LbGp (P < 0.05). Hypoxia-induced enhancement of intracellular calcium ([Ca2+]i) was attenuated by LbGp significantly (P < 0.01). Moreover, KCl-induced enhancement of [Ca2+]i was also reduced by LbGp at the doses of 25, 50, 100 microg x mL(-1) in a concentration-dependent manner. CONCLUSION: The result suggested that LbGp is able to increase the survival ratio and inhibit the enhancement of the intracellular free calcium concentration in cardiomyocytes induced by hypoxia and high potassium. One of the mechanisms is that LbGp acts on L-type calcium channels.

[Chemistry of polysaccharide Lzps-1 from Ganoderma lucidum spore and anti-tumor activity of its total polysaccharides].

AIM: To study the structure and anti-tumor activity of polysaccharide from Ganoderma lucidum spore treated with microwave. METHODS: DEAE-cellulose and Sephadex G-50 column chromatography were used to isolate and purify the polysaccharide whose structure was characterized by using chemical and spectral methods. RESULTS AND CONCLUSION: One polysaccharide, named Lzps-1 was obtained from the water extract, with its molecular weight estimated by HPGPC to be 8000. Its structure was investigated to be glucan. The total polysaccharides, Lzps processed antitumor activity against sarcoma 180 and Lewis lung cancer in mice and enhanced the NK cell activity. Lzps-1 is obtained for the first time from Ganoderma spore Lzps has anti-tumor activity.

[Physical-chemical properties and structure elucidation of abPS isolated from the root of Achyranthes bidentata].

AIM: To study the physicochemical properties and the structure of Achyranthes bidentata polysaccharide (AbPS). METHODS: AbPS was isolated from the roots of Achyranthes bidentata Bl., and purified by gel filtration chromatography. The distribution of the molecular weight of AbPS was determined by ESI-MS. The structure of AbPS was deduced by methylation analysis, reductive-cleavage and 13CNMR spectroscopy. RESULTS: AbPS was shown to compose of fructose residues and glucose residues and the molar ratio was 8:1. AbPS contain 2,1-linked fructose residue, 2,1-linked fructose residue, 1,2,6-linked fructose residue, terminal fructose residue and terminal glucose residue. CONCLUSION: AbPS is a fructan and belong to graminan.

[Studies on physico-chemical properties and hypoglycemic activity of complex polysaccharide AMP-B from Atractylodes macrocephala Koidz].

AIM: To isolate a complex polysaccharide (AMP-B) from Atractylodes macrocephala Koidz and study its phtsico-chemical properties and hypoglycemic activity. METHODS: The root of Atractylodes macrocephala K. was extracted with water and precipitated with ethanol, dialyzed against water and freeze-dried to get the crude polysaccharides (AMP). A complex polysaccharide (AMP-B) was isolated and purified on DEAE-cellulose column. The model of diabetes rats was established with alloxan injection through the tail vein. Male rats were divided into 5 groups: the normal group, the control group, and three AMP-B-fed groups. Measuring the blood glucose, water and food consumption, thymus and pancreas index, and studying cut sections of pancreas tissues. RESULTS: AMP-B is a complex-polysaccharide, elemental analysis of AMP-B shown C 32.84%, H 5.68%, and N 1.79%. The neutral polysaccharide content of AMP-B was 50.3%, uronic acid was 40.4%, and protein was 11.5%. Monosaccharide composition of AMP-B was determined by GC, AMP-B composed of Glc, Gal, Man, Ara and Rha in a molar ratio of 3.0:2.5:1.3:3.5:1.0. AMP-B was found to reduce blood glucose level in alloxan-diabetic rats markedly at doses of 50, 100 and 200 mg.kg-1 by ig, but no effect in normal rat. AMP-B was found to decrease the consumption of water and food, recover pancreas damage of diabetic rats obviously, inhibited the atrophy of thymus and pancreas of the diabetic rats induced by alloxan. CONCLUSION: AMP-B showed significant hypoglycemic effect on the experimental hyperglycemias rats induced by alloxan.

Structural elucidation and antitumor activity of a fructan from Cyathula officinalis Kuan.

A fructan named CoPS3 was isolated from Cyathula officinalis Kuan. The structure of CoPS3 was determined by methylation, by the reductive-cleavage method combined with GC-MS analysis, and both 1D and 2D 1H and 13C NMR spectroscopy. These results show that CoPS3 is a graminans-type fructan that is comprised of a beta-D-fructofuranosyl backbone having residues linked (2-->1)- and (2-->6) with branches and an alpha-D-glucopyranose residue on the nonreducing end of the fructan chain. Each branch is terminated by a beta-D-Fruf residue. Bioassay showed that it could inhibit growth of Lewis pulmonary carcinoma implanted in mice.

Synthesis of a targeting drug for antifibrosis of liver; a conjugate for delivering glycyrrhetin to hepatic stellate cells.

The neoglycoproteins that consist of human serum albumin (HSA) modified with mannose 6-phosphate ([M6P]x-HSA) were synthesized, and they showed high binding property to hepatic stellate cells (HSC) by immunohistochemical analysis. In addition, an increased substitution (X) of 6-phosphated mannose (M6P) was associated with an increased accumulation in HSC. So the [M6P]x-HSA might be a carrier to deliver drugs to HSC. The antifibrotic drug, glycyrrhetin, was chosen to conjugate to M6P(26)-HSA. The result suggests there were 6 approximately 7 glycyrrhetin molecules having been conjugated to the carrier. Targeting glycyrrhetin to HSC might reduce its adverse affects and increase the efficacy.