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1.
Orapin Insuan Phornphimon Janchai Benchaluk Thongchuai Rujirek Chaiwongsa Supaporn Khamchun Somphot Saoin Wimonrut Insuan Peraphan Pothacharoen Waraporn Apiwatanapiwat Antika Boondaeng Pilanee Vaithanomsat 《Current issues in molecular biology》2021,43(1):93
Bromelain is a mixture of proteolytic enzymes derived from pineapple (Ananas comosus) fruit and stem possessing several beneficial properties, particularly anti-inflammatory activity. However, the molecular mechanisms underlying the anti-inflammatory effects of bromelain are unclear. This study investigated the anti-inflammatory effects and inhibitory molecular mechanisms of crude and purified rhizome bromelains on lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells. RAW264.7 cells were pre-treated with various concentrations of crude bromelain (CB) or purified bromelain (PB), and then treated with LPS. The production levels of pro-inflammatory cytokines and mediators, including nitric oxide (NO), interleukin (IL)-6, and tumor necrosis factor (TNF)-α were determined by Griess and ELISA assays. The expressions of inducible nitric oxide synthetase (iNOS), cyclooxygenase (COX)-2, nuclear factor kappa B (NF-κB), and mitogen-activated protein kinases (MAPKs)-signaling pathway-related proteins were examined by western blot analysis. The pre-treatment of bromelain dose-dependently reduced LPS-induced pro-inflammatory cytokines and mediators, which correlated with downregulation of iNOS and COX-2 expressions. The inhibitory potency of PB was stronger than that of CB. PB also suppressed phosphorylated NF-κB (p65), nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha, extracellular signal-regulated kinases, c-Jun amino-terminal kinases, and p38 proteins in LPS-treated cells. PB then exhibited potent anti-inflammatory effects on LPS-induced inflammatory responses in RAW264.7 cells by inhibiting the NF-κB and MAPKs-signaling pathways. 相似文献
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Korakot Nganvongpanit Peraphan Pothacharoen Patama Chaochird Kasisin Klunklin Kanawee Warrit Jongkolnee Settakorn Nuttaya Pattamapaspong Sirichai Luevitoonvechkij Olarn Arpornchayanon Prachya Kongtawelert Dumnoensun Pruksakorn 《Arthritis research & therapy》2009,11(3):R78
Introduction
The purpose of this study was to evaluate serum chondroitin sulfate (CS) and hyaluronic acid (HA) levels and the capability of cartilage repair of full-thickness cartilage defects after treatment with two different fundamental surgical techniques: autologous chondrocyte transplantation (AC) and subchondral drilling (SD).Methods
A 4-mm-diameter full-thickness cartilage defect was created in each of 10 skeletally mature male outbred dogs. The dogs were randomly separated into two groups. Groups A and B were treated with AC and SD, respectively. An evaluation was made at the 24th week of the experiment. Serum was analyzed prospectively – preoperatively and at 6-week intervals – for CS and HA levels by enzyme-linked immunosorbent assay (ELISA) and ELISA-based assays, respectively.Results
The cartilage repair assessment score (median ± standard deviation) of group A (9.5 ± 2.5) was significantly higher than that of group B (2.5 ± 1.3) (P < 0.05). Group A also demonstrated a better quality of hyaline-like cartilage repair. Prospective analysis of serum WF6 and HA levels between the two groups did not show any significant difference. Serum WF6 levels at the 24th week of the experiment had a negative correlation (r = -0.69, P < 0.05) with the cartilage repair assessment score, whereas serum HA levels tended to correlate positively (r = 0.46, 0.1 <P < 0.05).Conclusions
AC treatment provides superior results to SD treatment, according to morphology, histology, and cartilage marker levels. AC treatment demonstrated a smoother surface, less fissure, better border integration, and a more reliable outcome of repairing cartilage. Moreover, a decreasing level of serum WF6, which correlated with good quality of the repairing tissue at the end of the follow-up period, was found predominantly in the AC group. Serum WF6 therefore should be further explored as a sensitive marker for the noninvasive therapeutic evaluation of cartilage repair procedures. 相似文献4.
Itthiarbha A Phitak T Sanyacharernkul S Pothacharoen P Pompimon W Kongtawelert P 《In vitro cellular & developmental biology. Animal》2012,48(1):43-53
Interleukin-1β (IL-1β) induces the expression of matrix metalloproteinases (MMPs) implicated in cartilage and joint degradation
in osteoarthritis (OA) and rheumatoid arthritis (RA). Polyoxypregnane glycoside (PPG), active compound was identified from
Dregea volubilis extract by chemical analysis, shown to exert chondroprotective effects in cartilage explant models. However, no studies have
been undertaken for the molecular investigation of whether PPG constituents protect the human articular chondrocyte (HAC).
In the present studies, HAC was co-treated with IL-1β and PPG. The expression of MMPs, type II collagen, phosphorylation of
mitogen-activated protein kinases (MAPKs) and NF-κB signaling pathway were determined by Western immunoblotting. PPG (6.25–25 μM)
decreased the IL-1β-induced HA release from chondrocyte to culture medium. The mode of action of PPG was likely mediated through
inhibiting expression of MMP-1, -3 and -13 in the medium, which was associated with the inhibition of mRNA expression. PPG
had no effect on IL-1β-induced phosphorylation of MAPK pathway. Conversely, PPG decreased phosphorylation of IκB kinase and
IκBα degradation. Taken together, these results indicate that PPG may inhibit cartilage degradation in OA and may also be
used as nutritional supplement for maintaining joint integrity and function. 相似文献
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Pothacharoen P Kalayanamitra K Deepa SS Fukui S Hattori T Fukushima N Hardingham T Kongtawelert P Sugahara K 《The Journal of biological chemistry》2007,282(48):35232-35246
Chondroitin sulfate (CS) proteoglycans are major components of cartilage and other connective tissues. The monoclonal antibody WF6, developed against embryonic shark cartilage CS, recognizes an epitope in CS chains, which is expressed in ovarian cancer and variably in joint diseases. To elucidate the structure of the epitope, we isolated oligosaccharide fractions from a partial chondroitinase ABC digest of shark cartilage CS-C and established their chain length, disaccharide composition, sulfate content, and sulfation pattern. These structurally defined oligosaccharide fractions were characterized for binding to WF6 by enzyme-linked immunosorbent assay using an oligosaccharide microarray prepared with CS oligosaccharides derivatized with a fluorescent aminolipid. The lowest molecular weight fraction recognized by WF6 contained octasaccharides, which were split into five subfractions. The most reactive subfraction contained several distinct octasaccharide sequences. Two octasaccharides, DeltaD-C-C-C and DeltaC-C-A-D (where A represents GlcUAbeta1-3GalNAc(4-O-sulfate), C is GlcUAbeta1-3Gal-NAc(6-O-sulfate), D is GlcUA(2-O-sulfate)beta1-3GalNAc(6-O-sulfate), DeltaCis Delta(4,5)HexUAalpha1-3GalNAc(6-O-sulfate), and DeltaDis Delta(4,5)HexUA(2-O-sulfate)alpha1-3GalNAc(6-O-sulfate)), were recognized by WF6, but other related octasaccharides, DeltaC-A-D-C and DeltaC-C-C-C, were not. The structure and sequences of both the binding and nonbinding octasaccharides were compared by computer modeling, which revealed a remarkable similarity between the shape and distribution of the electrostatic potential in the two different octasaccharide sequences that bound to WF6 and that differed from the nonbinding octasaccharides. The strong similarity in structure predicted for the two binding CS octasaccharides (DeltaD-C-C-C and DeltaC-C-A-D) provided a possible explanation for their similar affinity for WF6, although they differed in sequence and thus form two specific mimetopes for the antibody. 相似文献
6.
Deer antler is a highly regenerative tissue that involves cellular differentiation, osteogenesis and ossification processes.
Chondroitin sulfate is the major glycosaminoglycan contained in antler connective tissue and has been isolated from cartilaginous
antler by 4 M GuHCl extraction, gradient ultracentrifugation and chromatography techniques. We examined the disaccharide composition
by 2-AB labeling and anion exchange HPLC analysis of the three resultant fractions (high, medium and low density fractions).
The high density fraction consists of A-unit and D-unit disaccharide in the ratio of 1:1, whereas, the CS disaccharide composition
ratio of A- unit:C-unit:D-Unit:E-unit contained in medium and low density fractions are 3:4:3:1 and 2:2:2:1, respectively.
The only intact CS oligosaccharides of the medium density fraction upregulated gene expression of bone-specific proteins of
a human osteoblastic cell line (hFOB1.19). Thus, CS oligosaccharides from cartilaginous deer antler, with their oversulfated
chondroitin sulfate composition, demonstrated the physiological properties and may be good candidates for osteogenetic agents
in humans. 相似文献
7.
Peraphan Pothacharoen Sumet Najarus Jongkolnee Settakorn Shuji Mizumoto Kazuyuki Sugahara Prachya Kongtawelert 《Glycoconjugate journal》2014,31(3):221-230
Osteoarthritis (OA) is a degenerative joint disease that progressively causes a loss of joint functions and the impaired quality of life. The most significant event in OA is a high degree of degradation of articular cartilage accompanied by the loss of chondroitin sulfate-proteoglycans (CS-PGs). Recently, the chondroprotective effects of sesamin, the naturally occurring substance found in sesame seeds, have been proved in a rat model of papain-induced osteoarthritis. We hypothesized that sesamin may be associated with possible promotion of the biosynthesis of CS-PGs in human articular chondrocytes. The aim of the study was to investigate the effects of sesamin on the major CS-PG biosynthesis in primary human chondrocyte. The effects of sesamin on the gene expression of the PG core and the CS biosynthetic enzymes as well as on the secretion of glycosaminoglycans (GAGs) in monolayer and pellet culture systems of articular chondrocytes. Sesamin significantly increased the GAGs content both in culture medium and pellet matrix. Real-time-quantitative PCR showed that sesamin promoted the expression of the genes encoding the core protein (ACAN) of the major CS-PG aggrecan and the biosynthetic enzymes (XYLT1, XYLT2, CHSY1 and CHPF) required for the synthesis of CS-GAG side chains. Safranin-O staining of sesamin treated chondrocyte pellet section confirmed the high degree of GAG accumulation. These results were correlated with an increased level of secreted GAGs in the media of cultured articular chondrocytes in both culture systems. Thus, sesamin would provide a potential therapeutic strategy for treating OA patients. 相似文献
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Peraphan Pothacharoen Kanyamas Choocheep Tanyaluck Pitak Wilart Pompimon Bhusana Premanode Timothy E. Hardingham Prachya Kongtawelert 《Central European Journal of Biology》2006,1(3):430-450
We investigated the effects of A. galanga extract on metabolism and gene expression involved in the interleukin-1β (IL-1β) response of human chondrocyte and synovial fibroblast. A. galanga extract inhibited IL-1β enhanced matrix breakdown of the cartilage explants in a dose-dependent manner. It suppressed uronic acid loss from the tissue and decreased the release of sulfated GAG and hyaluronan into the medium. MMP-2 and MMP-9 activity in the culture medium of chondrosarcomas and synovial fibroblasts were significantly reduced in the presence of A. galanga extract, which also suppressed the production of MMP-1,-3 and-13. The A. galanga extract also significantly increased type II collagen, SOX9 and aggrecan gene expression, suggesting an ability to enhance anabolic activity. At a high dose of A. galanga extract there was a down-regulation of aggrecan gene expression. Comparison with Diacerein® showed its general anti-inflammatory potential to be similar. The A. galanga extract was shown to inhibit IL-1β-stimulated cartilage matrix degradation in both systems. Additionally, the extract showed the potential to up-regulate certain chondrocyte anabolic genes. It may, therefore, offer some cartilage protective and anti-inflammatory properties as a therapeutic agent in arthritis. 相似文献
9.
Wichaya Sriuttha Nantawat Uttamo Apisek Kongkaew Jongkolnee Settakorn Suchanan Rattanasalee Prachya Kongtawelert Dumnoensun Pruksakorn Peraphan Pothacharoen 《Cell and tissue banking》2016,17(4):721-734
Due to the inconvenient and invasive nature of chondrocyte transplantation, preserved cartilage has been recognized as an alternative source of chondrocytes for implantation. However, there are major concerns, in particular, the viability and quality of the chondrocytes. This study investigated the biochemistry and molecular characterization of chondrocytes isolated from preserved cartilage for purposes of transplantation. Ex vivo characterization was accomplished by storing human cartilage at either 4 or ?80 °C in a preservation medium. Microscopic evaluation of the preserved cartilage was conducted after 1, 2, 3 and 6 weeks. The chondrocytes were isolated from the preserved cartilage and investigated for proliferation capacity and chondrogenic phenotype. Transplantation of chondrocytes from preserved cartilage into rabbit knees was performed for purposes of in vivo evaluation. The serum cartilage degradation biomarker (WF6 epitopes) was evaluated during the transplantation procedure. Human cartilage preserved for 1 week in a 10 % DMSO chondrogenic medium at 4 °C gave the highest chondrocyte viability. The isolated chondrocytes showed a high proliferative capacity and retained chondrogenic gene expression. Microscopic assessment of the implanted rabbit knees showed tissue regeneration and integration with the host cartilage. A decreased level of the serum biomarker after transplantation was evidence of in vivo repair by the implanted chondrocytes. These results suggest that cartilage preservation for 1 week in a 10 % DMSO chondrogenic medium at 4 °C can maintain proliferation capacity and the chondrogenic phenotype of human chondrocytes. These results can potentially be applied to in vivo allogeneic chondrocyte transplantation. Allogeneic chondrocytes from preserved cartilage would be expected to maintain their chondrogenic phenotype and to result in a high rate of success in transplanted grafts. 相似文献
10.
Pothacharoen P Siriaunkgul S Ong-Chai S Supabandhu J Kumja P Wanaphirak C Sugahara K Hardingham T Kongtawelert P 《Journal of biochemistry》2006,140(4):517-524
OBJECTIVE: To determine the value of serum chondroitin sulfate epitope WF6 and hyaluronan (HA) levels as a biomarker for early detection of ovarian epithelial cancer and other gynecological disorders. METHOD: Serum WF6 CS epitope and HA were measured in 91 patients with ovarian epithelial cancer, 39 patients with non-cancer gynecological disorders and 30 healthy women. Serum chondroitin sulfate (CS) WF6 epitope was determined by a competitive immunoassay with the monoclonal antibodies WF6, which specifically recognizes an epitope in native CS chains. In addition, serum HA concentration was measured by an ELISA-based assay with a biotinylated affinity HA-binding proteins. RESULTS: The serum concentration of CS (WF6) epitope was highly increased in epithelial types of ovarian cancer and at all stages of development (p < 0.005). Serum HA in ovarian cancer patients was significantly higher than normal controls (p < 0.05). CONCLUSION: These results reflect changes in ECM metabolism in progressive ovarian cancer, which cause an increase in serum CS epitopes and HA. Therefore, serum CS epitopes may provide useful biomarkers for cancers and other disorders of the ovary. Measurement of serum HA provided complementary information, which may be useful as a discriminator between benign ovarian disorders and malignant ovarian diseases. 相似文献