소 태아 혈청이 연골의 Proteoglycan 합성에 미치는 영향

김흥준,김재도,김영옥,허만하 고신대학교(의대) 고신대학교 의과대학 학술지 1994 고신대학교 의과대학 학술지 Vol.10 No.1

Articular cartilage is a highly specialized connective tissue that covers the ends of long bones within the synovial joint cavity. It contains no blood vessels, no nerve fibers, and no lymphatics, accordingly it is nourished from the synocial flued by diffusion. Articular cartilage is composed of and extracellular matrix(ECM), collagen and proteoglycan. A small number of specific cells, chondrocytes, are embedded in the matrix. They are influenced by the alteration of components in synovial fluid which is derived from serum. To examine the effect of serum on biosynthesis of proteoglycan, this experiment was performed using human articular cartilages which were obtained from the patients who were operated due to various diseases. The human articular cartilages were incubated in 3 kinds of media : DMEM without fetal calf serum(FCS) (control), DMEM containing 1% FCS, and DMEM containing 20% FCS. The synthethized proteoglycan, incorporated with labeled S³⁵ sulfate, was extracted through PD-10 column after digestion by papain and measured the uptake of [35S] sulfate labeled proteoglycan by scintillation counter. To examine whether difference in concentration of fetal calf serum is reflected in staining intensity of immunohistochemistry for proteoglycan(avidin-biotin peroxidase complex), immunohistochemical staining was performed. The results were as followings : 1) Biosyntheses of proteoglycan in 1% and 20% FCS were increased 1.77 times and 2.27 respectively comparing with the control group. 2) In FCS group, biosynthesis of proteoglycan in 20% FCS was 1.29 times higher than that in 1% FCS. 3) No difference in staining intensity for proteoglycan in the variable concentrations of fetal calf serum was seen. The above results suggest that the compositional alteration of synovial fluid due to various factors influence the synthesis of the ECM in the articular cartilage and immunohistochemical staining is not sensitive enough to differentiate the quantitative difference of antigen.

소 태아 혈청이 연골의 Proteoglycan 합성에 미치는 영향

김흥준,김재도,김영옥,허만하 고신대학교 의학부 1994 高神大學校 醫學部 論文集 Vol.10 No.1

Articular cartilage is a highly specialized connective tissue that covers the ends of long bones within the synovial joint cavity. It contains no blood vessels, no nerve fibers, and no lymphatics, accordingly it is nourished from the synovial fluid by diffusion. Articular cartilage is composed of an extracellular matrix(ECM), collagen and proteoglycan. A small number of specific cells, chondrocytes, are embedded in the matrix. They are influenced by the alteration of components in synovial fluid which is derived from serum. To examine the effect of serum on biosynthesis of proteoglycan, this experiment was performed using human articular cartilages which were obtained from the patients who were operated due to various diseases. The human articular cartilages were incubated in 3 kinds of media : DMEM without fetal calf serum(FCS) (control), DMEM containing 1 % FCS, and DMEM containing 20 % FCS. The synthethized proteoglycan, incorporated with labeled S^(35) sulfate, was extracted through PD-10 column after digestion by papain and measured the uptake of [35S] sulfate labeled proteoglycan by scintillation counter. To examine whether difference in concentration of fetal. calf serum is reflected in staining intensity of immunohistochemistry for proteoglycan (avidin-biotin peroxidase complex), immunohistochemical staining was performed. The results were as followings : 1) Biosyntheses of proteoglycan in 1 % and 20% FCS were increased 1.77 times and 2.27 respectively comparing with the control group. 2) In FCS groups, biosynthesis of proteoglycan in 20% FCS was 1.29 times higher than that in 1% FCS. 3) No difference in staining intensity for proteoglycan in the variable concentrations of fetal calf serum was seen. The above results suggest that the compositional alteration of synovial fluid due to various factors influence the synthesis of the ECM in the articular cartilage and immunohistochemical staining is not sensitive enough to differentiate the quantitative difference of antigen.

상황버섯 추출물에 의한 인체혈구암세포의 Telomerase 활성 저하

홍수현,한민호,김혜정,최영현 대한암예방학회 2010 Journal of cancer prevention Vol.15 No.2

Recent studies demonstrated that proteoglycan extracted from Phellinus linteus has a potent anti-tumor and/or anti-inflammatory activity by leading to induction of apoptosis and inhibition of prostaglandin E2production; however, its mechanism of action and its molecular targets on the telomere length regulation in human cancer remain unclear. In this study, we investigated the effect of proteoglycan on the levels of telomere regulatory components and the activity of telomerase in human leukemia U937 cells. Exposure of U937 cells to proteoglycan resulted in growth inhibition and induction of apoptosis as measured by MTT assay and flow cytometry analysis, which was associated with induction of tumor suppressor p53and cyclin-dependent kinase inhibitor p21 (WAF1/CIP1). Proteoglycan treatment markedly inhibited the activity of telomerase, and the levels of and transcription factor c-myc, human telomerase reverse transcriptase (hTERT) and telomerase-associated protein-1 (TEP-1), main determinants of the telomerase enzymatic activity, were progressively down-regulated by proteoglycan treatment in a dose-dependent fashion. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of proteoglycan. (Cancer Prev Res 15, 172-177, 2010)

상황버섯 추출물에 의한 인체혈구암세포의 세포사멸 및 Prostaglandin E2의 생성 저하

박철,현숙경,황혜진,최성현,최영현 대한암예방학회 2010 Journal of cancer prevention Vol.15 No.1

Various mushroom polysaccharides and polysaccharides-protein complexes are known to possess anti-tumor and immunomodulating effects. Here, we demonstrated that proteoglycan extracted from Phellinus linteus has a potent anti-tumor and/or anti-inflammatory activity. Proteoglycan treatment resulted in a concentration-dependent growth inhibition by including apoptosis, which could be proved by DAPI staining, agarose gel electrophoresis and flow cytometry analysis. The increase in apoptosis induced by proteoglycan was correlated with up-regulation of pro-apoptotic Bax and Bad expression, and downregulation of anti-apoptotic Bcl-2 and Bcl-xL expression. Proteoglycan inhibited the levels of IAP family members such as XIAP and cIAP-1 and induced the proteolytic activation of caspase-3, and a concomitant degradation of β-catenin. In addition, it was found that proteoglycan treatment markedly decreased the levels of cyclooxygenases (COX)-2 mRNA and protein expression without significant changes in the expression of COX-1, which was correlated with a decrease in prostaglandin E2 synthesis. Taken together,these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of proteoglycan. (Cancer Prev Res 15, 83-91, 2010

The Effects of Proteoglycan and Type II Collagen on T1ρ Relaxation Time of Articular Cartilage

최원석,류혜진,홍성환,최자영 대한영상의학회 2015 대한영상의학회지 Vol.72 No.2

Purpose: To evaluate the effects of proteoglycan and type II collagen within articular cartilage on T1ρ relaxation time of articular cartilage. Materials and Methods: This study was exempted by the institutional and animal review boards, and informed consent was not required. Twelve porcine patellae were assigned to three groups of control, trypsin-treated (proteoglycan-degraded), or collagenase-treated (collagen-degraded). The T1ρ images were obtained with a 3 tesla magnetic resonance imaging scanner with a single loop coil. Statistical differences were detected by analysis of variance to evaluate the effects of the enzyme on T1ρ relaxation time. Safranin-O was used to stain proteoglycan in the articular cartilage and immunohistochemical staining was performed for type II collagen. Results: Mean T1ρ values of the control, trypsin-treated, and collagenase-treated groups were 37.72 ± 5.82, 57.53 ± 8.24, and 45.08 ± 5.31 msec, respectively (p < 0.001). Histology confirmed a loss of proteoglycan and type II collagen in the trypsin- and collagenase-treated groups. Conclusion: Degradation of proteoglycans and collagen fibers in the articular cartilage increased the articular cartilage T1ρ value.

Isolation and Characterization of Proteoglycan Derived From Human Placenta and its Biological Activities

Lee, Kyung-Bok,Kim, Jong-Sig,Yoo, Yung-Choon,Kwak, Sang-Tae,Song, Kyung-Sik,Kim, Yeong-Shik The Pharmaceutical Society of Korea 2000 Archives of Pharmacal Research Vol.23 No.2

Chondroitin sulfates proteoglycans were isolated from human placenta. For the identification of enzymatic digestion products of isolated proteoglycan, strong anion exchange-high performance liquid chromatography (SAX-HPLC) was performed. By the action of chondroitin ABC and chondroitin B lyase, three unsaturated disaccharides 2-acetamide-2-deoxy-3-O-($\beta$-D-gluco-4-enepyranosyluronic acid)-D-galactose ($\delta$Di-OS), 2-acetamide-2-deoxy-3-O-($\beta$-D-gluco-4-enepyranosyluronic acid)-6-O-su lfo-D-galactose ($\delta$Di-6S) and 2-acetamide-2-deoxy-3-O-($\beta$-D-gl uco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose ($\delta$Di-4S) were produced from the human placenta proteoglycan. The anticoagulant activity of chondroitin sulfate proteoglycan was evaluated by activated partial thromboplastin time (aPTT) assay and thrombin time (TT) assay. The clotting times of aPTT and TT were increased from 72 to 144 sec and 19 to 27 sec, respectively. The Immune-modulating activity of chondroitin sulfate proteoglycan was examined by cell proliferation assay and these results suggest that it may play a role in suppression of the function of immune-related cells.

Microbial Subversion of Heparan Sulfate Proteoglycans

Ye Chen,Martin Götte,Jian Liu,Pyong Woo Park 한국분자세포생물학회 2008 Molecules and cells Vol.26 No.5

The interactions between the host and microbial pathogen largely dictate the onset, progression, and outcome of infectious diseases. Pathogens subvert host components to promote their pathogenesis and, among these, cell surface heparan sulfate proteoglycans are exploited by many pathogens for their initial attachment and subsequent cellular entry. The ability to interact with heparan sulfate proteoglycans is widespread among viruses, bacteria, and parasites. Certain pathogens also use heparan sulfate proteoglycans to evade host defense mechanisms. These findings suggest that heparan sulfate proteoglycans are critical in microbial pathogenesis, and that heparan sulfate proteoglycan-pathogen interactions are potential targets for novel prophylactic and therapeutic approaches.

Basement Membrane Proteoglycans: Modulators Par Excellence of Cancer Growth and Angiogenesis

Renato V. Iozzo,Jason J. Zoeller,Alexander Nystroem 한국분자세포생물학회 2009 Molecules and cells Vol.27 No.5

Proteoglycans located in basement membranes, the nanos-tructures underling epithelial and endothelial layers, are unique in several respects. They are usually large, elon-gated molecules with a collage of domains that share structural and functional homology with numerous extracellular matrix proteins, growth factors and surface receptors. They mainly carry heparan sulfate side chains and these contribute not only to storing and preserving the biological activity of various heparan sulfate-binding cytokines and growth factors, but also in presenting them in a more “active configuration” to their cognate receptors. Abnormal expression or deregulated function of these proteoglycans affect cancer and angiogenesis, and are critical for the evolution of the tumor microenvironment. This review will focus on the functional roles of the major heparan sulfate proteoglycans from basement membrane zones: perlecan, agrin and collagen XVIII, and on their roles in modulating cancer growth and angiogenesis.

마우스에서 IL-1ß가 염증의 발현에 미치는 영향에 관한 연구

윤덕상(Duk-Sang Yoon),이기수(Ki-Soo Lee) 대한치과교정학회 1998 대한치과교정학회지 Vol.28 No.4

이 연구는 염증반응에서 IL-β의 생성을 관찰하고, IL-β가 조직에 미치는 효과를 관찰하여 염증반응에서 IL-β의 역할을 검토할 목적으로 시행되었다. 실험동물은 웅성과 자성 마우스 각각 100마리를 대상으로 하였으며, 다음의 3단계 실험을 수행하였다. 1. 리포다당(lipopolysaccharide, LPS)에 의한 염증발현 시스템을 확립하기 위한 면역 B-세포에 대한 영향평가, 세포독성 없이 최대의 IL-β를 생성할 수 있는 LPS의 양 결정 및 LPS 투여로 생성되는 조직의 IL-β를 동정하는 실험의 결과, LPS는 B-림포세포의 활성을 농도 의존적으로 높였으며, 10-50μg 투여로 세포독성이 없이 IL-β를 최대로 생성함을 나타냈고, 여러 장기의 조직에서 IL-β를 생성시키나 특히, 구강조직과 관철활핵세포에서 많은 양을 생성함을 확인하였다. 2. 관절강에 50μg의 LPS를 투여하고 IL-β와 TNF α의 경시적 생성량 변화를 관찰하는 실험에서, IL-β는 투여 2시간 후에 증가되기 시작하여 4-5시간 경과 후에 최고농도에 도달하고, 점차 감소하여 24시간 경과 후에는 미량만이 존재하였으며, TNF α는 투여 1시간 경과 후에 증가되기 시작하여 2-3시간 경과 후에 최고농도에 도달하고, 그 이후에 감소되기 시작하여 6시간 경과후에는 미량만이 검출되었다. 3. IL-β를 정상조직에 투여하여 IL-β의 효과를 관찰한 실험에서, 관절강에 투여하여 leucocyte의 수가 투여 4-5시간 경과후에 최고치에 도달하고 36시간 경과 후에 대조군 수치에 도달하였으며, 세포기질인 proteoglycaqn의 소실은 15시간 경과 후 시작되어 45-60시간 경과 후에 최대이었고, 90시간 경과 후에 대조군 수치로 복귀함이 관찰되었으며, 구강조직에 투여하여 cyclooxigenase 대사산물(prostandin E₂)의 축적이 일정농도 한계에서는 농도 의존적으로 증가함을 나타냈다. 이상의 결과로부터 LPS로 유도된 마우스 관절염 활액에서 IL-β는 염증반응 초기의 수 시간이내에 생성되어 4-5시간 경과후에 최대양이 생산되며, 염증 조직에서 leucocyte(염증세포)의 침윤, proteoglycan의 소실 및 cyclooxygenase 대사산물(PGE₂)의 축적을 유도하는, 즉 염증반응을 중재하는 사이토카인 중의 하나임이 확이되었다. Cytokines are intercellular peptide mediators that regulate homeostasis and host defense reactions in living body. Of the diversity of cytokines in terms of biological accomplishment, interleukin 1-S ( IL-1β) and tumor necrosis factor(TNF) are the most conspicuous cytokines with a wide variety of effects on cells involved in inflammatory and immune responses, and likely to be involved in the inflammatory pathogenesis of oral tissue as well. present study was designed to explicate the role of IL-1β on inflammatory revelation of oral tissues in mice biochemically. In the Induced arthritis by injection of 10#956;g LPS shown the relaese of 0.93#956;g IL-β/joint with a peak at 4-5 h and diminished at 24h, and the release of TNF a of 1,25#956;g/joint with a peak at 2-3h and diminished at 6h. After injection of th IL-β into the joint, the member of leucocytes proliferated with a peak at 4-5h and diminished at 36h and the loss of proteoglycan showed with maximum at 15-30h. After injection of IL-1β into the oral tissue, cycloosygenase metabolites (PGEz) accumulated in the oral tissue with dose dependant. These elucidated IL-1β to be inflammatory mediator in the early phase of its pathogenesis. Intraoral injection of recombinant IL-1β induced the proliferation of leukocytes in situ. IL-1β took an pertinent part in the development of inflammation and the succession of cellular infiltration. The results exemplify that IL-1β plays a significant role in mediating inflammatory response induced by LPS in oral tissue the inflammatory response is regulated by IL-1β at an acute phase of pathogenesis.

배양한 백서의 사구체 상피세포에 대한 당과 후기당화합물에 의한 heparan sulfate proteoglycan의 변화

하태선(Tae Sun Ha),(Balakuntalam S Kasinath),김헌식(Hun Sik Kim) 대한신장학회 2000 Kidney Research and Clinical Practice Vol.19 No.1

N/A HSPG, a component of size-and charge-selective barrier of glomerular basement membrane, is one of important matrix proteins which has been known to be reduced in the kidney of diabetic patients or animals. To examine the effects of glucose and AGE on the HSPG production by cultured GEC, we cultured rat GEC on the AGE- or BSA-coated plate under normal(5mM) and high glucose.(30mM) conditions and measured the change of HSPG production by sandwich-ELISA assay and northern blot analysis at 2 days and one week incubation periods. There was no difference in proliferation between 2 different conditions of culture plate surface. We measured the relative amount of the extracted HSPC and observed significant decreases in high glucose condition at one week incubation, and particularly on the AGE-coated surface as compared to the results of BSA-coated condition, by 22% and 5%, respectively. The expression of mRNA for perlecan promoter was decreased in condition of high glucose and AGE-coated surface by 20Yo at 2 days and 61i at one week. Even in normal glucose condition, the expression of mRNA was reduced by 30Yo at one week if the plate was coated with AGE. In conclusion, both high glucose and AGE have reducing effects on the production of HSPG by GEC in vitro. Their effects seem to be additive, however, the role of AGE is greater than that of glucose, This means that the effort to inhibit AGE formation is more important than short-term glucose control for the prevention of diabetic proteinuria.