• 제목/요약/키워드: trypsin activity

검색결과 404건 처리시간 0.025초

치은섬유아세포와 치은인대세포의 형태와 화학주성에 미치는 대조추출물의 효과에 관한 연구 (A STUDY OF THE EFFECT OF ZIZYPHUS FRUCTUS EXTRACTS ON MORPHOLOGY & CHEMOTAXIS OF GINGIVAL FIBROBLAST & PERIODONTAL LIGAMENT CELLS)

  • 양창호;류인철;최상묵;정종평
    • Journal of Periodontal and Implant Science
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    • 제25권2호
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    • pp.279-289
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    • 1995
  • The most important object of periodontal treatment is the perfect regeneration of destructed periodontal tissue. The healing of periodontal lesion is affected by several cells & factors, which result in formation of long juntional epithelium, root resorption, bony ankylosis or connective tissue attachment. And ideal healing is enhanced by epithilial exclusion or periodontal ligament cell activation. In this investigation, I studied the effect of Zizyphus Fructus extract which enhances biologic activity& collagen synthesis, on the chemotaxis & cell nature. The cells were obtained from interdental area & middle third area of the freshly extracted teeth for the orthodontic purpose. And they were fully incubated in${\alpha}-MEM$ solution containing $100{\mu]g/ml$ penicillin & $100{\mu]g/ml$ streptomycin followed by 6 generation incubation. The test cells were collected by trypsin-EDTA & centrifuge in the fully incubated cells, counted by Hernacyotmeter, incbated $5{\times}10^5/ml$ cells for 24 hours, re-incubated 24 hours in media containing natural extract and photographed. The cells were incubated for 4 hours in 48 well microchemotaxis chamber bisecting upper & lower chamber by 8ug/m pore polycarbonate membrane coating 5mg/ml gelatin solution. The migrated cells in microscope were counted, which meaned cell chemotaxis activity. The study had shown that the morphology of cell was spindle-shaped as the control group, and the subextract test groups were not significantly different. In gingival fibroblasts, the chemotaxis effect of PDGF was statistically significant compared to control group. The Zizyphus Fructus extract was more or less enhanced chemotaxis effect and in $1{\mu}g/ml$ concentration the chemotaxis effect was slightly elevated compared with $10{\mu}g/ml$ concentration. But, among the subextracts, it was not significantly defferent. In PDL cells, the chemotaxis effect of PDGF in statistically significant, and the zizyphus Fructus extract had shown the enhanced effect. The effect was slightly higher in $1{\mu}g/ml$ concentration than 10g/ml concentration,and no significance among the subextracts.

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Effects of Heat-oxidized Soy Protein Isolate on Growth Performance and Digestive Function of Broiler Chickens at Early Age

  • Chen, X.;Chen, Y.P.;Wu, D.W.;Wen, C.;Zhou, Y.M.
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권4호
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    • pp.544-550
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    • 2015
  • This study was conducted to investigate effects of heat-oxidized soy protein isolate (HSPI) on growth performance, serum biochemical indices, apparent nutrient digestibility and digestive function of broiler chickens. A total of 320 1-day-old Arbor Acres chicks were randomly divided into 4 groups with 8 replicates of 10 birds, receiving diets containing soy protein isolate (SPI, control group) or the same amount of SPI heated in an oven at $100^{\circ}C$ for 1, 4, or 8 h, for 21 days, respectively. The results indicated that compared with the control group, body weight gain and feed intake of birds fed diet containing SPI heated for 8 h were significantly lower (p<0.05). Serum urea nitrogen concentration was higher in the broilers fed diet containing SPI heated for 4 or 8 h at d 21 (p<0.05). In contrast, serum glucose content was decreased by HSPI substitution at d 21 (p<0.05). The relative pancreas weight in HSPI groups was higher than that in the control group at d 21 (p<0.05). Meanwhile, the opposite effect was observed for relative weight of anterior intestine and ileum in broilers fed a diet containing SPI heated for 8 h (p<0.05). Birds fed diets containing SPI heated for 4 or 8 h had a decreased lipase activity in anterior intestinal content at d 14 and 21 (p<0.05), respectively. In addition, the same effect was also noted in broilers given diets containing SPI heated for 1 h at d 21 (p<0.05). Similarly, amylase, protease and trypsin activity in anterior intestinal content were lower in broilers fed diets containing SPI heated for 8 h at d 21 (p<0.05). The apparent digestibility of dry matter (DM) from d 8 to 10 and DM, crude protein (CP), and ether extract from d 15 to 17 were lower in broilers fed diets containing SPI heated for 8 h (p<0.05). Besides, birds given diets containing SPI heated for 4 h also exhibited lower CP apparent digestibility from d 15 to 17 (p<0.05). It was concluded that HSPI inclusion can exert a negative influence on the growth performance of broilers, which was likely to result from the simultaneously compromised digestive function.

Purinergic Receptors Play Roles in Secretion of Rat von Ebner Salivary Gland

  • Kim, Sang-Hee;Cho, Young-Kyung;Chung, Ki-Myung;Kim, Kyung-Nyun
    • International Journal of Oral Biology
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    • 제31권4호
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    • pp.141-148
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    • 2006
  • The effects of adenosine triphosphate(ATP) on salivary glands have been recognized since 1982. The presence of purinergic recepetors(P2Rs) that mediate the effects of ATP in various tissues, including parotid and submandibular salivary gland, has been supported by the cloning of receptor cDNAs and the expression of the receptor proteins. P2Rs have many subtypes, and the activation of these receptor subtypes increase intracellular $Ca^{2+}$, a key ion in the regulation of the secretion in the salivary gland. The apical pores of taste buds in circumvallate and foliate papillae are surrounded by the saliva from von Ebner salivary gland(vEG). Thus, it is important how the secretion of vEG is controlled. This study was designed to elucidate the roles of P2Rs on salivary secretion of vEG. Male Sprague-Dawley rats (about 200 g) were used for this experiment. vEG-rich tissues were obtained from dissecting $500-1,000\;{\mu}m$ thick posterior tongue slices under stereomicroscope view. P2Rs mRNA in vEG acinar cells were identified with RT-PCR. To observe the change in intracellular $Ca^{2+}$ activity, we employed $Ca^{2+}-ion$ specific fluorescence analysis with fura-2. Single acinar cells and cell clusters were isolated by a sequential trypsin/collagenase treatment and were loaded with $10\;{\mu}M$ fura -2 AM for 60 minutes at room temperature. Several agonists and antagonists were used to test a receptor specificity. RT-PCR revealed that the mRNAs of $P2X_4$, $P2Y_1$, $P2Y_2$ and $P2Y_3$ are expressed in vEG acinar cells. The intracellular calcium activity was increased in response to $10\;{\mu}M$ ATP, a P2Rs agonist, and 2-MeSATP, a $P2Y_1$ and $P2Y_2R$ agonist. However, $300\;{\mu}M\;{\alpha}{\beta}-MeATP$, a $P2X_1$ and $P2X_3R$ agonist, did not elicit the response. The responses elicited by $10\;{\mu}M$ ATP and UTP, a $P2Y_2R$ agonists, were maintained when extracellular calcium was removed. $10\;{\mu}M$ suramin, a P2XR antagonist, and reactive blue 2, a P2YR antagonist, partially blocked ATP-induced response. However, when extracellular calciums were removed, suramin did not abolish the responses elicited by ATP. These results suggest that P2Rs play an important role in salivary secretion of vEG acinar cells and the effects of ATP on vEG salivary secretion may be mediated by $P2X_4$, $P2Y_1$, $P2Y_2$, and/or $P2Y_3$.

비병원성 야생효모 Candida oleophila WP5-19-1로부터 장내유용세균 유해 효소 β-glucuronidase 저해제의 생산 및 특성 (Production and Characterization of the Beneficial β-glucuronidase Inhibitor from non pathogenic wild yeast, Candida oleophila WP5-19-1, and its effects on gut microbes )

  • 박선정;이종수
    • 한국균학회지
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    • 제50권1호
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    • pp.31-40
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    • 2022
  • 비병원성 야생효모로부터 대장에서의 독성물질 활성화에 관여하여 대장암을 유발시키는 효소 중의 하나인 β-glucuronidase를 저해하는 물질을 생산하기 위하여 대전광역시 월평공원으로부터 분리 동정한 야생효모들 중 비병원성 야생효모들을 선별하여 배양 상등액과 무세포 추출물을 각각 제조한 후 β-glucuronidase 저해활성을 측정하였다. 133균주의 비병원성 야생효모들 중 C. oleophila WP5-19-1의 무세포추출물이 49.0%의 가장 높은 저해활성을 보여 우수 균주로 최종 선발하였다. β-Glucuronidase 저해물질 최적 생산 조건은 C. oleophila WP5-19-1를 초기 pH를 6.0으로 조정한 dextrose 5% 함유한 potato dextrose 배지에 접종하여 30℃에서 24시간 배양하였을 때였고 이때 IC50값; 8.4 mg으로 가장 높은 저해활성을 보였다. C. oleophila WP5-19-1의 β-glucuronidase 저해물질을 한외여과와 Sephadex G-50으로 gel 여과를 실시하여 부분 정제하였고 이들은 30-60℃, pH 6.0-9.0에서 80% 이상의 잔존 저해활성을 보여 안정하였다. 또한 철, 구리, 아연과 같은 중금속 이온들은 저해활성을 활성화시켰다.

항생제 대체 사료첨가제로서 매실추출 혼합물이 육계의 성장, 소화 효소 활성도 및 장내 미생물 균총에 미치는 영향 (Effects of a Blend of Prunus Mume Extract as an Alternative to Antibiotics on Growth Performance, Activity of Digestive Enzymes and Microflora Population in Broiler Chickens)

  • 고영현;양혜영;강선영;김은숙;장인석
    • Journal of Animal Science and Technology
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    • 제49권5호
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    • pp.611-620
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    • 2007
  • 1950년대 이후 어린 가축의 성장 촉진과 사료효율을 개선하고 축산물의 생산효율을 향상시킬 목적으로 penicillin, bacitracin 등과 같은 항생제가 사료첨가제로서 널리 사용되어 왔다. 항생제의 작용기전은 먼저 가축체내에 존재하는 살모넬라, 대장균, 클로스트리디움 등과 같은 장내 병원성균의 세포막을 분해하여 사멸시키고(Leitner 등, 2001) 소장 흡수세포벽을 얇게 하여 영양소의 흡수율을 증대시켜 성장을 촉진시킨다고 알려져 있다(Ravindran 등, 1984; Collington 등, 1990). 그러나 가축 생산성 증대와 같은 긍정적인 효과에도 불구하고 항생물질이 고기, 우유, 계란 등의 축산물에 잔류될 수 있다는 점과 축산물을 통하여 섭취된 항생물질이 인체의 내성을 증가시켜 항생물질에 대하여 저항성을 나타낼 수 있다는 일부 부정적인 효과가 밝혀지면서 최근 항생제의 사용에 문제점이 제기되고 있다(Lee 등, 2001). 스웨덴, 덴마크 등과 같은 유럽 국가들을 중심으로 가축 사료첨가제로서 항생제 사용이 금지 또는 극도로 제한되고 있다. 최근 이와 같은 시대적 흐름에 따라 사료에 첨가되는 항생제를 53종에서 25종으로 대폭 축소하고, 2012년 농장에 대한 HACCP 제도를 도입하는 등 사료 위생관리를 엄격히 관리하는 단계로 접어들면서, 항생제를 대치 할 수 있는 대체제제에 대한 연구가 절실히 요구된다. 따라서 축산식품 소비자는 물론 생산자를 위해서도 사료에 첨가되는 항생제를 대체할 수 있는 안전성이 확보된 후보 물질의 연구 개발 및 효능 연구는 미래 지속가능한 동물산업을 위해 필연적으로 해결해야만 하는 향후의 숙제이다. 이러한 사실과 더불어 국민소득이 증가함에 따라 웰빙에 대한 관심 고조로 동물성 식품에 대한 선호도가 양 보다는 질 위주로 급격하게 변화되고 있어 천연자원을 이용한 다양한 항생제 대체 사료첨가제의 개발이 주목을 받고 있는바, 본 연구에서 사용된 매실(Prunus mume)은 섬유소, 탄닌과 무기물이 풍부하고 시트릭산(구연산) 등을 포함한 각종 유기산이 다량 함유된 것으로(임, 1999), 한방과 민간에서 잎, 청매(미성숙 매실)는 살균, 구충, 해독, 해열 등과 같은 다양한 효과를 나타내는 한약제로 널이 이용되고 있다. 매실의 효능은 다양한 연구에서 항균작용(이 등, 2003), 항산화작용(한 등, 2001), 간 기능 증진(서 등, 1990) 등의 효과들이 과학적으로 검정되었다. 특히 매실의 항균작용 연구에서 매실추출물로서 paper disc법으로 항균작용을 조사한 결과 그람(+) 세균인 Micrococcus leteus, Bacillus subtilis, Staphy- lococcus aureus 등과 그람(-) 세균인 E. coli. Salmonella typhimurium, Proteus vulgaris 등에 효과적인 저해 활성을 나타내었다(임, 1999; 이 등, 2003). 또한 유기산의 일종은 유산(lactic acid)은 이미 위장관의 환경을 긍정적으로 조절하여 가축 사료첨가제로서 상당히 많이 이용되고 있다(van de Broek, 2000). 그러나 장관내 환경조절물질로서 고품질의 축산물을 생산할 수 있는 천연 소재인 매실 또는 그 부산물을 이용하여 가축 사료첨가제로서의 가능성을 과학적으로 연구한 사례는 극히 일부분에 지나지 않고 있다. 따라서 본 연구는 매실추출 혼합물의 in vitro 항균효과를 검정하고 이를 근거로 육계사료에 위장관 환경조절제로서 매실추출물 및 유산(lactic aicd) 그리고 항산화제로서 포도씨앗추출물을 혼합한 제제를 급여하여 성장 및 사료효율, 혈액성상, 소장의 소화 효소 활성도 및 세균총에 미치는 작용에 대하여 조사하여 항생제 대체제로서의 가능성에 대한 연구를 실시하였다.

담배거세미나방에 살충효과를 나타내는 새로운 Bacillus thuringiensis 균주의 특성 (Characterization of New Bacillus thuringiensis Isolated with Bioactivities to Tobacco Cutworm, Spodoptera litura (Lepidoptera: Noctuidae))

  • 김다아;김진수;길미라;백승경;최수연;김대용;윤용남;황인천;유용만
    • 한국응용곤충학회지
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    • 제47권1호
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    • pp.87-93
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    • 2008
  • 국내 토양으로부터 분리한 Bacillus thuringiensis 균주에서 담배거세미나방(Spodoptera litura)에 선택적으로 살충효과를 나타내는 새로운 균주를 선발하였다. 이 균주의 결정성 독소단백질은 위상차 현미경과 주사전자현미경으로 관찰한 결과 전형적인 이중피라미드 형태이며 H serotype으로 동정한 결과 Bacillus thuringiensis subsp. CAB 109로 동정되었다. 난방제 해충인 담배거세미나방 3령충에 대해 B. t. subsp. aizawai CAB 109균주외 분리된 다른 3균주의 생물활성을 비교 검토하였다. 살충활성 결과에서 B. t. subsp. aizawai CAB 109 균주가 $1.3{\times}10^7$ (cfu/ml)에서 100% 사망률로 다른 균주에 비해 높은 활성을 나타내었다. 담배거세미나방은 배추좀나방과 다르게 7일 이상 경과한 뒤에야 100% 사충률을 나타내었다. 한편, 담배거세미나방 2령, 3령, 4령충에 대한 살충활성의 검정에서 B. t. aizawai subsp. CAB 109 균주의 $LD_{50}$값이 각각 $9.78{\times}10^5,\;6.87{\times}10^6,\;1.83{\times}10^7$ (cfu/ml)으로 살충활성을 나타내었다. B. t.를 섭식한 담배거세미나방 유충의 사망 현상은 지효성이며 대조군보다 무게가 $6{\sim}7$배정도 적으며 다음 령기로 성장하지 못하고 죽었다. 또한 B. t. subsp. aizawai CAB 109 균주의 SDS-PAGE에서의 단백질 패턴과 trypsin을 처리한 단백질의 결과에서 기존의 B. t. subsp. aizawai와 CAB 109균주와의 차이점은 발견할 수 없었다.

소의 수란관액에 의한 사람 난포액의 Gelatinase A 동위효소인 GA110의 분해 (A Gelatinase A Isoform, GA110, of Human Follicular Fluid Is Degraded by the Bovine Oviductal Fluid Component)

  • 김민정;김지영;이승재;윤용달;조동재;김해권
    • 한국발생생물학회지:발생과생식
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    • 제5권1호
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    • pp.23-33
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    • 2001
  • 포유류의 난자가 수란관내로 배란될 때는 난포액 성분도 같이 수란관내로 들어간다. 본 연구에서는 처음으로 난포액의 일부 성분이 수란관액에 의해서 변화하는 것을 관찰하였다. 사람의 난포액을 gelatin zymogram으로 분석한 결과 621kDa gelatinase 이외에 110kDa gelatinase (GA110) 등의 여러 gelatinase 활성이 나타났다. 이 활성들은 EDTA나 phenanthroline에 의해 억제된 반면 PMSF 처리에 의해서는 아무런 변화가 없었다. 소의 수란관액에서는 62kDa gelatinase의 활성만이 주로 관찰되었다. 소의 수란관 액과 사람의 난포액을 1:1로 섞고 이를 37$^{\circ}$C에서 3시간 동안 둔 결과 사람의 난포액의 GA110 활성은 사라졌다. 사람의 난포액에 APMA를 첨가한 결과 GA110의 활성은 대부분 감소하고 대신 62kDa gelatinase의 활성은 오히려 증가하였다. 반면에 사람의 난포액에 EDTA를 3시간 동안 처리한 결과 GA110의 활성은 오히려 현저히 증가하였고 이 때 다른 gelatinases의 활성은 영향을 받지 않았다. PMSF나SBTI는 난포액내의 gelatinases활성에 아무런 변화를 일으키지 않았다. EDTA, PMTA 혹은 SBTI 등의 proteinase inhibitor를 미리 처리한 사람의 난포액에 소의 수란관 내액을 섞은 경우에도GA110의 활성은 여전히 감소하였다. 사람의 혈청에서도 EDTA에 의해 활성이 현저히 증가하는 GA110이 발견되었다. 사람의 난포액과 유사하게 혈청내의 GA110도 소의 수란관액에 의하여 활성이 사라졌다. 그러나 사람의 난포과립세포의 추출물에서는 단지 92kDa gelatinase만 관찰이 되었다. 마지막으로 anti-human gelatinase A 항체를 사용하여 사람의 난포액과 혈청 그리고 난포과립세포의 추출액을 western blotting한 결과 621kDa과 GA110 만이 항원-항체 반응을 나타내었다. 이 같은 결과로 미루어 사람의 난포액과 혈청에는 gelatinase A의 독특한 isoform인 GA110이 있으며 특히 난포액내의 GA110은 수란관액성분에 의해 선택적으로 분해되는 것으로 여겨진다.

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Effects of a lipid-encapsulated zinc oxide supplement on growth performance and intestinal morphology and digestive enzyme activities in weanling pigs

  • Jang, Insurk;Kwon, Chang Hoon;Ha, Duck Min;Jung, Dae Yun;Kang, Sun Young;Park, Man Jong;Han, Jeong Hee;Park, Byung-Chul;Lee, Chul Young
    • Journal of Animal Science and Technology
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    • 제56권8호
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    • pp.29.1-29.6
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    • 2014
  • This study compared the effects of varying lipid content and dietary concentration of a lipid-encapsulated (LE) ZnO product to those of native ZnO and thereby to find insights into optimal lipid coating and dosage of the Zn supplement. A total of 192 21-d-old weanling pigs were allotted to 48 pens, after which each six pens received a ZnO-free basal diet supplemented with 125 ppm ZnO (100 ppm Zn; BASAL), 2,500 ppm Zn as native ZnO (HIGH), or 100 or 200 ppm Zn as LE ZnO (LE-100 or LE-250) containing 8%, 10%, or 12% lipid [LE-8%, LE-10%, or LE-12%, respectively; $2{\times}3$ factorial arrangement within the LE-ZnO diets (LE-ALL)] for 14 d. Forty pigs were killed at the end for histological and biochemical examinations. None of ADG, ADFI, gain:feed, and fecal consistency score differed between the LE-ALL and either of the BASAL and HIGH groups. Hepatic and serum Zn concentrations were greater (p <0.05) in the HIGH vs. LE-ALL group, but did not differ between LE-ALL and BASAL, between LE-100 and -250, or among LE-8%, -10%, and -12% groups. Villus height (VH), crypt depth (CD), and the VH:CD ratio in the duodenum, jejunum, and ileum did not differ between the LE-ALL and either of the BASAL and HIGH groups, except for a greater CD in the duodenum in the LE-ALL vs. HIGH group. Additionally, VH and CD in the duodenum and VH:CD in the jejunum were greater in the LE-250 vs. LE-100 group. Specific activities of sucrase, maltase, and leucine aminopeptidase in these intestinal regions and those of amylase and trypsin in the pancreas were not influenced by the lipid content or dietary concentration of LE ZnO and also did not differ between the LE-ALL and either of the BASAL and HIGH groups, except for a greater pancreatic amylase activity in the former vs. HIGH group. In conclusion, the present results indicate that the LE ZnO, regardless of its lipid percentage or supplementation level examined in this study, has no significant effect on growth performance, fecal consistency, or digestive enzyme activities of weanling pigs under the experimental conditions.

Determination of human breast cancer cells viability by near infrared spectroscopy

  • Isoda, Hiroko;Emura, Koji;Tsenkova, Roumiana;Maekawa, Takaaki
    • 한국근적외분광분석학회:학술대회논문집
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    • 한국근적외분광분석학회 2001년도 NIR-2001
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    • pp.4105-4105
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    • 2001
  • Near infrared spectroscopy (NIRS) was employed to qualify and quantify on survival, the injury rate and apoptosis of the human breast cancer cell line MCF-7 cells. MCF-7 cells were cultured in RPMI medium supplemented with 10% FCS in a 95% air and 5% CO2 atmosphere at 37$^{\circ}C$. For the viable cells preparation, cells were de-touched by 0.1% of trypsin treatment and washed with RPMI supplemented with 10% FCS medium by centrifugation at 1000 rpm for 3min. For the dead cells preparation, cells were de-touched by a cell scraper. The cells were counted by a hemacytometer, and the viability was estimated by the exclusion method with frypan blue dye. Each viable and dead cells were suspended in PBS (phosphate bufferred saline) or milk at the cell density desired. For the quantitative determination of cell death by measuring the LDH (lactate dehydrogenase) activity liberated from cells with cell membrane injuries, LDH-Cytotoxic Test Wako (Wako, Pure Pharmaceutical Co. Ltd., Japan) was used. We found that NIRS measurement of MCF-7 cells at the density range could evaluate and monitor the different characteristics of living cells and dead cells. The spectral analysis was performed in two wavelength ranges and with 1,4, 10 mm pathlength. Different spectral data pretreatment and chemometrics methods were used. We applied SIMCA classificator on spectral data of living and dead cells and obtained good accuracy when identifying each class. Bigger variation in the spectra of living cells with different concentrations was observed when compared to the same concentrations of dead cells. PLS was used to measure the number of cells in PBS. The best model for measurement of dead cells, as well as living cells, was developed when raw spectra in the 600-1098 nm region and 4 mm pathlength were used. Smoothing and second derivative spectral data pretreatment gave worst results. The analysis of PLS loading explained this result with the scatter effect found in the raw spectra and increased with the number of cells. Calibration for cell count in the 1100-2500 nm region showed to be very inaccurate.

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수종의 cytokine이 사람 치주인대 섬유아세포의 prostaglandine $E_2$, leukotriene $B_4$ 및 collagenase 생산에 미치는 영향 (EFFECT OF VARIOUS CYTOKINES ON THE PRODUCTION OF PROSTAGLANDIN $E_2$, LEUKOTRIENE $B_4$ AND COLLAGENASE IN HUMAN PERIODONTAL LIGAMENT FIBROBLASTS IN VITRO)

  • 김정호;서정훈
    • 대한치과교정학회지
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    • 제24권4호
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    • pp.871-883
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    • 1994
  • This experiment was designed to study possible roles of $interleukin-1\beta$, interleukin-6 and tumor necrosis $factor-\alpha$ in bone remodeling by measuring their effects on $PGE_2,\; LTB_4$ and collagenase production when they were administered to human periodontal ligament fibroblasts. Human periodontal ligament fibroblasts were collected from first premolars extracted for orthodontic treatment. They were incubated in the environment of $37^{\circ}C,\;5\%\;Co^2,\;and\;100\%$ humidity. They were treated with $0.25\%$ trypsin-EDTA solution and centrifuged. PDL cells in the fifth to seventh passage were used for the experiment. Cells were seeded onto the culture dishes and when they were successfully attached, human recombinant $interleukin-1\beta$, interleukin-6, and tumor necrosis $factor-\alpha$ were administered, alone or in combination. They were incubated for 4, 8 and 24 hours and the levels of $PGE_2,\;LTB_4$ and collagenase released into the culture media were assessed by enzymeimmunoassay and collagenase activity assay. The conclusions are as follows: 1. $IL-1\beta\;and\;TNF-\alpha$ were very active in stimulating the production of $PGE_2$ and collagenase by human periodontal ligament fibroblasts, while IL-6 increased $LTB_4$ production. 2. $IL-1\beta$ significantly increased $PGE_2$, but $LTB_4$ Production was not increased. $IL-1\beta$ is thought to act mainly via the cyclooxygenase pathway of arachidonic acid metabolism. 3. IL-6 tended to inhibit $IL-1\beta$ in the production of $PGE_2$ and collagense whereas IL-6 and $TNF-\alpha$ showed auditive effect in the level of $PGE_2$. The above cytokines increased the release of at least one of $PGE_2,\;LTB_4$ and collagenase. It suggests that cytokines are involved in bone remodeling process by stimulating PDL fibroblasts to produce various bone-resorptive agents. The roles of cytokines in bone remodeling as a whole would need further study.

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