Guangxin Yuan

Anti-diabetic effect of Coptis Chinensis polysaccharide in high-fat diet with STZ-induced diabetic mice

For the past few years, numerous polysaccharides and polysaccharide-protein complexes have been isolated from plant or animal and used as a promising source of therapeutic agents for diabetes mellitus (DM). In this study, a water-soluble polysaccharide, named as CCPW-1, was extracted and fractioned from the roots of Coptis Chinensis by DEAE Sepharose Fast Flow anion-exchange and Sephadex G-100 column chromatography. The determination of the monosaccharide composition in CCPW-1 with gas chromatography (GC) showed that CCPW-1 was composed of glucose (54.8%), arabinose (22.3%), xylose (11.5%), galactose (7.6%) and galacturonic acid (3.8%). Diabetic mice induced by high-fat diet (HFD) with streptozotocin (STZ) were administered CCPW-1 (100, 50, 25 mg/kg b.w.). Effects of CCPW-1 on bodyweight, fasting blood glucose (FBG), oral glucose tolerance test (OGTT), fasting serum insulin (FINS), total glycerin (TG), total cholesterol (TC), super oxygen dehydrogenises (SOD), catalase (CAT) and methane dicarboxylic aldehyde (MDA) were investigated. CCPW-1 could improve the bodyweight, reduce the content of FBG and enhance FINS level. Meanwhile, CCPW-1 significantly suppressed the rise in blood glucose after 30 min in OGTT. TG and TC levels of diabetic mice also decreased after CCPW-1 treatment. Furthermore, CCPW-1 showed an obvious antioxidant effect through increasing SOD, CAT activities and decreasing MDA content in pancreas. These results indicate that CCPW-1 could be developed to a potent drug used for the treatment of DM in the future.

The contribution of side chains to antitumor activity of a polysaccharide from Codonopsis pilosula

In order to assess the potential contribution of the side chains of a polysaccharide (CPPW1) from the roots of Codonopsis pilosula to the biological activities, we chose CPPW1 and its backbone (CPPW1B) as the research targets to compare them with their antitumor and immunomodulatory activity. The results demonstrated CPPW1 could significantly inhibit the tumor growth of H22-bearing mice and simulate lymphocyte proliferation. Meanwhile the phagocytic capability of macrophages and NO production were also enhanced. However, CPPW1B could only inhibit the tumor weight and did not work to immune system at all. Additionally, both CPPW1 and CPPW1B had no toxicity in vivo. Taken together with all data, we found the sugar side chains attached to the backbone of CPPW1 played a pivotal role in fighting against tumor and activating immune system.

Effects of compound K on hyperglycemia and insulin resistance in rats with type 2 diabetes mellitus.

Compound K (CK) is a final metabolite of panaxadiol ginsenosides from Panax ginseng. Although anti-diabetic activity of CK has been reported in recent years, the molecular mechanism of CK in the treatment of diabetes mellitus remains unclear. In the present investigation, we established a rat model of type 2 diabetes mellitus (T2DM) with insulin resistance using high-fat diet (HFD) and streptozotocin (STZ), and attempted to verify more details and exact mechanisms in the treatment of T2DM. CK was administered orally at three doses [300, 100 and 30 mg/kg bodyweight (b.w.)] to the diabetic rats. Bodyweight, food-intake, fasting blood glucose (FBG), fasting serum insulin (FINS), insulin sensitivity (ISI), total glycerin (TG), total cholesterol (TC), as well as oral glucose tolerance test (OGTT) were evaluated in normal and diabetic rats. According to our results, CK could improve bodyweight and food-intake of diabetic rats. CK exhibited dose-dependent reduction of FBG, TG and TC of diabetic rats. CK treatment also enhanced FINS and ISI. Meanwhile, the glucose tolerance observed in the present study was improved significantly by CK. It is concluded from the results that CK may have improving effects on hyperglycemia and insulin resistance of diabetic rats. Furthermore, research showed that CK could promote the expression of InsR, IRS1, PI3Kp85, pAkt and Glut4 in skeletal muscle tissue of diabetic rats. These results indicate that the hypoglycemic activity of CK is mediated by improvement of insulin sensitivity, which is closely related to PI3K/Akt signaling pathway.

Purification and physicochemical properties of different polysaccharide fractions from the water extract of Boschniakia rossica and their effect on macrophages activation

Today more and more attentions had been attracted by many nutritionists and pharmacologists on polysaccharides from natural plants or animals due to their significant biological activities. In this research three polysaccharides (BRR-W1, BRR-WA1 and BRR-WA2) were isolated and purified from the water extract of Boschniakia rossica by DEAE Sepharose Fast Flow and Sepharose 6 Fast Flow column chromatography. Chemical and physical characteristics of three polysaccharides were investigated by a combination of chemical and instrumental analysis methods. The assays of their effect on macrophages activation were also investigated in vitro, including phagocytosis of macrophages, detections for NO production and TNF-α secretion. The results indicated that the effect of polysaccharides on macrophages activation was influenced by their respective physicochemical properties.

Simultaneous and Rapid Determination of Main Lignans in Different Parts of Schisandra Sphenanthera by Micellar Electrokinetic Capillary Chromatography

Lignans are imporant active ingredients of Schisandra sphenanthera. A micellar electrokinetic chromatography method was developed for the simultaneous determination of eight lignans – schizandrin, schisandrol B, schisantherin A, schisanhenol, anwulignan, deoxyschizandrin, schizandrin B and schizandrin C – in different parts of S. sphenanthera. The key factors for separation and determination were studied and the best analysis conditions were obtained using a background electrolyte of 10 mM phosphate-37.5 mM SDS-35% v/v acetonitrile (pH 8.0) at the separation voltage of 28 kV and detection at 214 nm, whereby the plant samples could be analyzed within 9.0 min. Analysis yielded good reproducibility (RSD between 1.19-2.28%) and good recovery (between 92.2-103.8%). The detection limits (LOD) and limit of quantification (LOQ) were within 0.4-1.2 mg/L and 1.5-4.0 mg/L. This method is promising to improve the quality control of different parts of S. sphenanthera.

In Vitro Antioxidant activities and anti-diabetic effect of a polysaccharide from Schisandra sphenanthera in rats with type 2 diabetes

A water soluble polysaccharide (SSPW1), with an average molecular weight of 191.18KD, was isolated and purified from the water extract of Schisandra sphenanthera by DEAE-52 and Sephadex G-100 column chromatography. SSPW1 contained 48.92% neutral sugar, 5.56% proteins and 42.83% uronic acid, and were composed of rhamnose, arabinose, mannose, galactose, glucose at a molar ratio of 13.52, 5.69, 3.92, 41.28 and 35.59. In vitro experiment showed that SSPW1 have a satisfactory scavenging effect on superoxide anion free radical, hydroxyl radical and DPPH free radical. In vivo experiment showed that SSPW1 could increase the body weight, improve the glucose tolerance, reduce FBG, and elevate the levels of FINS and the value of ISI in the diabetic rats. In addition, SSPW1 could reduce the MDA content, and increase GSH-PX, CAT and SOD activities. These results suggest that SSPW1 has both in vitro and in vivo antioxidant effect, which may be closely related to its antidiabetic effect.

Identification of velvet antler by random amplified polymorphism DNA combined with non-gel sieving capillary electrophoresis

Abstract Mitochondrial DNA of velvet antler was amplified with random amplified polymorphic DNA (RAPD) technique and the PCR products were detected with non-gel sieving capillary electrophoresis to establish a RAPD-HPCE method used for identifying the authenticity of velvet antler or it counterfeits. Factors that could affect the PCR amplification and capillary electrophoresis were optimized. Under the optimized conditions, namely, 20 mmol L−1 NaH2PO4–Na2HPO4–2 mmolL−1 EDTA buffer solution [0.8% (W/V) HPMC, 15 mmol L−1 TBAP and pH 7.3], −10 kV injection voltage and −8 kV separation voltage, Cervus nippon Temminck antler, Cervus elaphus Linnaeus antler, Rangifer tarandus antler, Cervus canadensis antler and Elaphurus davidianus antler were analyzed. The analysis on the similarity of obtained elctrophoretograms showed that there were significant differences in similarities of different velvet antlers, which could be used for the quick identification of the authenticity of velvet antler samples. It can be found that the technique of RAPD combined with HPCE is advantageous in rich polymorphism, high detection rate, simple and convenient performance, high efficiency, rapidness and sensitivity, indicating that it should be suitable for the quick identification of the authenticity of velvet antler samples.

Antidiabetic mechanism of Coptis chinensis polysaccharide through its antioxidant property involving the JNK pathway

Abstract Context: Antidiabetic activity of Coptis chinensis Franch (Ranunculaceae) polysaccharide (CCPW) has been reported. However, its molecular mechanism remains unclear. Objective: An attempt was made to further verify the antidiabetic activity of CCPW on type 2 diabetes mellitus (T2DM) and elucidate the mechanism of antidiabetic activity. Materials and methods: Male Wistar rats were fed with high-fat diet (HFD) and injected with streptozotocin (STZ) to generate a T2DM model. Effects of CCPW on fasting blood glucose (FBG), triglyceride (TG), total cholesterol (TC), glutathione (GSH), glutathione peroxidases (GSH-Px), superoxide dismutases (SOD), catalase (CAT), malondialdehyde (MDA), c-jun n-terminal kinase (JNK), phosphorylated insulin receptor substrate 1 (phospho-IRS1), phosphorylated phosphatidylinositol 3 kinase (phospho-PI3Kp85) and glucose transporter 4 (Glut4) were investigated. Results: FBG level of diabetic rats could be significantly inhibited by 51.2, 42.7, and 23.3% through administration of CCPW at doses of 200, 100, and 50 mg/kg b.w., respectively (p < 0.01). CCPW also could significantly reduce TG by 19.2, 12.1, and 7.4%, and TC by 24.2, 20.9, and 18.7%, respectively (p < 0.05 or p < 0.01). CCPW showed an obvious antioxidant effect through increasing GSH-Px, SOD, and CAT activities, and decreasing GSH and MDA contents (p < 0.05 or p < 0.01). Furthermore, CCPW could inhibit JNK and phospho-IRS1 expression and promote the expression of phospho-PI3Kp85 and Glut4 compared with those in the DM group (p < 0.05 or p < 0.01). Discussion and conclusion: CCPW can produce antidiabetic activity in rats with T2DM through its antioxidative effect, which is closely related to the JNK/IRS1/PI3K pathway.

Characteristics of PCR-SSCP and RAPD-HPCE methods for identifying authentication of Penis et testis cervi in Traditional Chinese Medicine based on cytochrome b gene.

Abstract The use of Penis et testis cervi, as a kind of precious Traditional Chinese Medicine (TCM), which is derived from dry deers testis and penis, has been recorded for many years in China. There are abundant species of deer in China, the Penis et testis from species of Cervus Nippon and Cervus elaphusL were authentic, others species were defined as adulterant (different subspecies of deer) or counterfeits (different species). Identification of their origins or authenticity becomes a key in controlling the herbal products. A modified column chromatography was used to extract mitochondrial DNA of dried deers testis and penis from sika deer (C. Nippon) and red deer (C. elaphusL) in addition to adulterants and counterfeits. Column chromatography requires for a short time to extract mitochondrial DNA of high purity with little damage of DNA molecules, which provides the primary structure of guarantee for the specific PCR; PCR-SSCP method showed a clear intra-specific difference among patterns of single-chain fragments, and completely differentiate Penis et testis origins from C. Nippon and C. elaphusL. RAPD-HPCE was based on the standard electropherograms to compute a control spectrum curve as similarity reference (R) among different samples. The similarity analysis indicated that there were significant inter-species differences among Penis et testis’ adulterant or counterfeits. Both techniques provide a fast, simple, and accurate way to directly identify among inter-species or intra-species of Penis et testis.

2-[(4-Bromophenyl)(4-fluorophenylamino)methyl]cyclohexanone

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