Bioactive polysaccharides isolated from the fruiting bodies of Phellinus sp.

Yan Yang,Jingsong Zhang,Yanfang Liu,Qingbo Wang,Qingjiu Tang,Shuai Zhou 한국버섯학회 2010 한국버섯학회 국제학술대회 Vol.6 No.-

Bioactive polysaccharides isolated from the fruiting bodies of Phellinus sp.

Yan Yang,Jingsong Zhang,Yanfang Liu,Qingbo Wang,Qingjiu Tang,Shuai Zhou 한국버섯학회 2010 한국버섯학회지 Vol.8 No.4

Phellinus sp. are assigned to the Basidiomycotina, Hymenomycetes, Aphyllophorales and Hymenochaetaceae, and have been shown containing various bioactive substances including triterpenoids, polysaccharides and flavones[1]. Traditional Chinese herbalists believe that Phellinus species are effective in treating many gynecopathic ailments[2] and are also reported to exhibit other pharmacological functions including tumor cell inhibition, antioxidant activity and anti-hepatic fibrosis effects[3]. Polysaccharides of Phellinus sp. has been reported to possess antitumor activities and inhibit tumor recrudescence and metastasis. There are little studies comparing the chemical composition and biological activities difference among polysaccharides from different Phellinus sp and little report about the pure polysaccharide structure analysis. In this study, eight kinds of crude polysaccharides were extracted from Phellinus fruit bodies and their chemical composition and bioactivities were researched. The polysaccharide and protein contents of eight crude polysaccharides had a certain extent differences. Monosaccharide composition and content of amino acids also existed some differences in eight crude polysaccharides. Eight different polysaccharides all showed enhancing splenocyte proliferation effect in vitro. PB-10P and JSHP had high cell proliferation rates with 50㎍/ml concentrations. The results indicated in some extent the immune activity of crude polysaccharides were correlation with the polysaccharide and protein content and composition of each sample. The crude polysaccharides of P. igniarius were further isolated and purified using DEAE Sepharose F. F. and gel-filtration chromatography (Sephacryl S-100-500 )repeatedly. Five water-soluble homogeneous polysaccharides (P60w1、P60s1、P1SP1、P10SP1and P100SP1) were obtained. Lack of absorption at 280 nm and 260 nm by UV scanning indicated that contained no protein and nucleic acid. HPLC produced a single symmetrical peak, indicated homogeneity and their molecular weigh were 1.71×104 Da、2.07×104 Da、1.48×104 Da、2.20×104 Da and 2.56×104 Da respectively. Structural of P60w-1 were determined using sugar and methylation analysis combined with 1H and 13C NMR spectroscopy, including COSY, TOCSY, NOESY, HSQC and HMBC experiments. The effect of P60w-1 on tumor growth was examined using subcutaneously transplanted H22 and Lewis Lung Carcinoma (LLC) tumor mouse models. Cyclophosphamide or Coriolus versicolor polysaccharopeptide served as positive controls in evaluating tumor response. Results showed that P60w-1 at the most effective dose of 100 mg/kg inhibited the growth of H22 and LLC by 48% and 37%, respectively.

Peroxisome Proliferator-Activated Receptor α Facilitates Osteogenic Differentiation in MC3T3-E1 Cells via the Sirtuin 1-Dependent Signaling Pathway

Gong, Kai,Qu, Bo,Wang, Cairu,Zhou, Jingsong,Liao, Dongfa,Zheng, Wei,Pan, Xianming Korean Society for Molecular and Cellular Biology 2017 Molecules and cells Vol.40 No.6

Type 2 diabetes mellitus (T2DM) is a chronic metabolic disease characterized by lack of insulin and high glucose levels. T2DM can cause bone loss and fracture, thus leading to diabetic osteoporosis. Promoting osteogenic differentiation of osteoblasts may effectively treat diabetic osteoporosis. We previously reported that Sirtuin 1 (Sirt1), a $NAD^+$-dependent deacetylase, promotes osteogenic differentiation through downregulation of peroxisome proliferator-activated receptor (PPAR) ${\gamma}$. We also found that miR-132 regulates osteogenic differentiation by downregulating Sirt1 in a $PPAR{\beta}/{\delta}$-dependent manner. The ligand-activated transcription factor, $PPAR{\alpha}$, is another isotype of the peroxisome proliferator-activated receptor family that helps maintain bone homeostasis and promot bone formation. Whether the regulatory role of $PPAR{\alpha}$ in osteogenic differentiation is mediated via Sirt1 remains unclear. In the present study, we aimed to determine this role and the underlying mechanism by using high glucose (HG) and free fatty acids (FFA) to mimic T2DM in MC3T3-E1 cells. The results showed that HG-FFA significantly inhibited expression of $PPAR{\alpha}$, Sirt1 and osteogenic differentiation, but these effects were markedly reversed by $PPAR{\alpha}$ overexpression. Moreover, siSirt1 attenuated the positive effects of $PPAR{\alpha}$ on osteogenic differentiation, suggesting that $PPAR{\alpha}$ promotes osteogenic differentiation in a Sirt1-dependent manner. Luciferase activity assay confirmed interactions between $PPAR{\alpha}$ and Sirt1. These findings indicate that $PPAR{\alpha}$ promotes osteogenic differentiation via the Sirt1-dependent signaling pathway.

Peroxisome Proliferator-Activated Receptor α Facilitates Osteogenic Differentiation in MC3T3-E1 Cells via the Sirtuin 1-Dependent Signaling Pathway

Kai Gong,Bo Qu,Cairu Wang,Jingsong Zhou,Dongfa Liao,Wei Zheng,Xianming Pan 한국분자세포생물학회 2017 Molecules and cells Vol.40 No.6

Type 2 diabetes mellitus (T2DM) is a chronic metabolic disease characterized by lack of insulin and high glucose levels. T2DM can cause bone loss and fracture, thus leading to diabetic osteoporosis. Promoting osteogenic differentiation of osteoblasts may effectively treat diabetic osteoporosis. We previously reported that Sirtuin 1 (Sirt1), a NAD+-dependent deacetylase, promotes osteogenic differentiation through downregulation of peroxisome proliferator-activated receptor (PPAR) γ. We also found that miR-132 regulates osteogenic differentiation by downregulating Sirt1 in a PPAR /δ-dependent manner. The ligand-activated transcription factor, PPAR , is another isotype of the peroxisome proliferator-activated receptor family that helps maintain bone homeostasis and promot bone formation. Whether the regulatory role of PPAR in osteogenic differentiation is mediated via Sirt1 remains unclear. In the present study, we aimed to de-termine this role and the underlying mechanism by using high glucose (HG) and free fatty acids (FFA) to mimic T2DM in MC3T3-E1 cells. The results showed that HG-FFA significantly inhibited expression of PPAR , Sirt1 and osteogenic differentiation, but these effects were markedly reversed by PPAR overexpression. Moreover, siSirt1 attenuated the positive effects of PPAR on osteogenic differentiation, suggesting that PPAR promotes osteogenic differentiation in a Sirt1-dependent manner. Luciferase activity assay confirmed interactions between PPAR and Sirt1. These findings indicate that PPAR promotes osteogenic differentiation via the Sirt1-dependent signaling pathway.

An Auto-inducible Expression System Based on the RhlI-RhlR Quorum-sensing Regulon for Recombinant Protein Production in E. coli

Rao Ben,Fan Jiying,Sun Jian’an,Truong Ngoc Tu,Sun Jing,Zhou Jingsong,Qiuyi,Shen Yaling 한국생물공학회 2016 Biotechnology and Bioprocess Engineering Vol.21 No.1

An artificial network which can accomplish recombinant protein synthesis guided by cell population in E. coli was constructed. The successful functioning of this network requires two plasmids, pWNB and pET. pWNB is responsible for production of T7 RNA polymerase, which controls pET; pET, in turn, regulates the production of target proteins. Several model proteins were tested and the results show that this E. coli system can be used to efficiently express various recombinant proteins. Since system contains T7 RNA polymerase production elements, it is transferable and applicable to well-characterized E. coli strains. Compared to the IPTG-induced system, an equal or greater amount of target protein can be obtained using this auto-inducible expression system in flasks and bioreactors. Our results suggest that it is a competitive alternative to other expression systems used in labs or for industrial applications.