• Title/Summary/Keyword: mechanism study

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Membrane Protein Alterations Associated with Anticancer Drug Resistance in Mouse Lymphoblastic Leukemia L1210 Cells (항암제에 내성인 생쥐의 백혈병세포 L1210의 세포막 단백질의 변환)

  • Kim, Seong-Yong;Son, Sung-Kweon;Kim, Jae-Ryong;Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
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    • v.10 no.2
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    • pp.432-444
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    • 1993
  • Multidrug resistance(MDR) phenotype is frequently observed in animal and human cancer cell lines selected for in vitro resistance to a single chemotherapeutic agent. It is characterized by the diminished drug accumulation and is related to the drug efflux mechanism in resistant cells. In the present study, adriamycin resistant cells(L1210-$AdR_6$ : $10^{-6}M$ adriamycin, $-AdR_5$ : $10^{-5}M$) and vincristine resistant cells (L1210-$VcR_7$ : $10^{-7}M$ vincristine, $-VcR_6$ : $10^{-6}M$) were produced from mouse lymphoblastic leukemia cell line L1210. Growth profiles of survived cells were observed for 5 days with MTT(thiazolyl blue) assay and resistance was compared with $IC_{50}$(drug concentration of 50% survival reduction in absorbance). Resistant cells proliferated more slowly than sensitive cell. Doubling times were 29.7hr in L1210, 68.7hr in L1210-$AdR_5$ and 58.2hr in $-VcR_6$. MDRs expressed as resistance factor were as follows, L1210-$AdR_5$ was 76.4 times for vincristine, L1210-$VcR_6$ was 96.4 times for adriamycin. The cell membrane proteins with three different M.W. were recognized to be related resistance, 220, 158, and 88 kd in L1210-$AdR_5$, 158, 140 and 88 kd in L1210-$VcR_6$ by SDS-PAG electrophoresis. Cell surface membrane proteins were identified by radio-iodination and autoradiogram, their molecular weights were 158, 72.8, and 42.4 Kd in L1210-$VcR_6$.

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Induction of Apoptosis by Ethanol Extract of Lythrum anceps (Koehne) Makino in Human Leukemia U937 Cells (인체백혈병 U937 세포에서 부처꽃 에탄올추출물에 의한 apoptosis 유도)

  • Jeong, Jin-Woo;Kim, Chul Hwan;Lee, Young-Kyung;Hwang, Yong;Lee, Ki Won;Choi, Kyung-Min;Kim, Jung Il
    • Korean Journal of Plant Resources
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    • v.33 no.4
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    • pp.279-286
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    • 2020
  • Purple loosestrife-Lythrum anceps (Koehne) Makino is a herbaceous perennial plant belonging to the Lythraceae family. It has been used for centuries in Korea and other Asian traditional medicine. It has been showed pharmacological effects, including anti-oxidant and anti-microbial effects. However, the mechanisms underlying its anti-cancer effect are not yet understood. In this study, we investigated the mechanism of apoptosis signaling pathways by ethanol extract of Lythrum anceps (Koehne) Makino (ELM) in human leukemia U937 cells. Treatment with ELM significantly inhibited cell growth in a dose-dependent manner by inducing apoptosis, as evidenced by the formation of apoptotic bodies (ApoBDs), DNA fragmentation and increased populations of sub-G1 ratio. Induction of apoptosis by ELM was connected with up-regulation of death receptor (DR) 4 and DR5, pro-apoptotic Bax protein expression and down-regulation of anti-apoptotic Bcl-2 protein, and inhibitor of apoptosis protein (IAP) family proteins, depending on dosage. This induction was associated with Bid truncation, mitochondrial dysfunction, proteolytic activation of caspases (-3, -8 and -9) and cleavage of poly(ADP-ribose) polymerase protein. Therefore, our data indicate that ELM suppresses U937 cell growth by activating the intrinsic and extrinsic apoptosis pathways, and thus may have applications as a potential source for an anti-leukemic chemotherapeutic agent.

Effect of Steviol β-Glucopyranosyl Ester on The Production of Nitric Oxide and Inflammatory Cytokines in RAW 264.7 Cells (Steviol β-Glucopyranosyl Ester가 RAW 264.7 세포의 산화질소 및 염증성 사이토카인 생성에 미치는 영향)

  • Jung, Heehoon;Cho, Uk Min;Hwang, Hyung Seo;Cho, Kun;Lee, Sang Rin;Kim, Moo Sung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.3
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    • pp.239-247
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    • 2018
  • Chronic inflammation is known to have effects on various diseases such as gout, cancer, dementia, atopic disease, and obesity. In addition, since some signal cascades involved in the development of inflammation are known to affect the damage and aging of the skin tissue, studies are being conducted actively to control the inflammation mechanism. In order to mitigate or prevent inflammatory response, a number of researches have been made to develop anti-inflammatory materials from some plants. In particular, Stevia rebaudiana produces steviol glycosides (SG), a natural sweetener with a distinctive flavor. Studies on some of SG have been shown to have anti-inflammatory activity. Researchers of this study expected that more SG also possess anti-inflammatory activity, besides stevioside, rebaudioside A, and steviol. In order to confirm this possibility, the researchers screened inhibition activity of various steviol glucosides for NO production in RAW 264.7 cell lines. As a result, steviol ${\beta}-glucopyranosyl$ ester (SGE) showed the highest inhibitory activity among steviol derivatives treated at the same molar concentration. In addition, we found that mRNA expression level of $interleukin-1{\alpha}$ ($IL-1{\alpha}$), $interleukin-1{\beta}$ ($IL-1{\beta}$), cyclooxygenase-2 (COX-2), nuclear factor kappa-light chain-enhancer of activated B cells ($NF-{\kappa}B$) and inducible nitric oxide synthase (iNOS) was also decreased in a dose-dependent manner. These results show that SGE inhibits anti-inflammatory activity and NO production in mouse macrophage RAW 264.7 cells. It was confirmed that SGE has potential to be applied as an anti-inflammatory material.

Effect of Korean Red Ginseng on Blood Pressure and Aortic Vascular(endothelial) Histological Changes in Rats (홍삼이 정상 흰쥐의 혈압 및 대동맥 조직 소견 변화에 미치는 영향)

  • Joo, Il-Woo;Sung, Kyung-Hwa;Park, Jung-Min;Lew, Jae-Hwan;Oh, Han-Jin
    • Journal of Ginseng Research
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    • v.32 no.4
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    • pp.324-331
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    • 2008
  • Introduction : Korean red ginseng has been shown to have an preventive effect on atheroma formation and enhancing effect on nitric oxide synthesis in endothelial cell inducing vasodilatation. However, there are few studies showing the effect of Korean red ginseng on blood pressure and vascular(endothelial) pathologic changes together. We designed this study to show changes of blood pressure and vascular pathologic findings with Korean red ginseng administration compared with Chinese red ginseng and control for 3 months in rats. Materials and methods : We studied the in vitro hypotensive effect and effect on vascular pathologic changes of Korean red ginseng compared with Chinese red ginseng and control. Rats were randomly assigned to three groups(Korean red ginseng, Chinese red ginseng and control) and evaluated by blood pressure and aortic vascular(endothelial) pathologic changes monthly during 3-month administration. All results were analyzed by paired T-test, ANOVA and post-hoc. Results : Blood pressure lowering effect was noted on Korean red ginseng and Chinese red ginseng compared to control. Especially, in Korean red ginseng group, hypotensive effect was showed in first and second month, but, in Chinese red ginseng group, it was just noted in first month. In case of vascular(aortic endothelial) pathologic finding, endothelial wall thickening and elastic fiber tearing were noted in Chinese red ginseng group compared with Korean red ginseng group and control with statistical significance.(p<0.05) Discussion : These results suggested Korean red ginseng could have more hypotensive effect and maintenance rather than Chinese red ginseng. And the difference of hypotensive effect between Korean red ginseng and Chinese red ginseng might has some association with difference of vascular pathologic findings in each group. However, further evaluation and research of other mechanism will be needed to convince this hypothesis.

Ag Ion Substituted HAp Coatings on Ti-6Al-4V Substrate by IBAD and It's Bactericidal Effect (Ti 합금표면의 항균성 HAp Coating에 관한 연구)

  • Jung, Moon-Young;Kim, Taik-Nam;Kim, Yun-Jong;Yim, Hyuk-Jun;Kim, Jong-Ock;Lim, Dae-young;Kim, Sun-Ok
    • The Journal of Engineering Research
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    • v.3 no.1
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    • pp.189-197
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    • 1998
  • Hyeroxyapatite(HAp) which has good biocompatibility was made by Wet Chemical Process. The surface of Ti-6Al-4V, coated with HAp by lon Beam Assisted Deposition (IBAD), was treated with 5ppm, 10ppm, 20ppm, and 100ppm of $AgNO_3$ solution. In this Ag impregnation process, $Ca^{2+}$ of HAp was substituted with $Ag^+$ of $AgNO_3$. In this study, the antimicrobial effect and biocompatibility of Ti-6Al-4V alloy which was coated with Ag-HAp were examined. The antimicrobial test was carried out with two kinds of bacteria(P. Aeruginosa, S. Epidermidis), which are highly infectious in a transplanting operation of implant materials. As a result of the test, it was observed that Ti-6Al-4V alloy which was treated by 20ppm of $AgNO_3$ solution has good biocompatibility. In order to observe the antimicrobial mechanism of $Ag^+$, E. coli which is the most common bacterium was treated by Ag-HAp. Then cell morphology of E. coli was observed by the transmission electron microscope(TEM). The destruction of cell wall and cytoplasm of E. coil were observed. A black spot appeared in the cytoplasm was analyzed by energy dispersive analysis X-ray (EDAX) and it showed a small amount of $Ag^+$. Thus, it was proved that $Ag^+$ destroys bacteria effectively and Ti-6Al-4V alloy which was impregnate with Ag ion show antimicrobial effect on infection bacteria.

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Structural Features of the Glandular Trichomes in Leaves of Carnivorous Drosera anglica Huds. (식충식물 긴잎끈끈이주걱 (Drosera anglica Huds.) 분비모의 구조적 특성)

  • Baek, Kyung-Yeon;Kim, In-Sun
    • Applied Microscopy
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    • v.38 no.1
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    • pp.21-28
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    • 2008
  • Carnivorous plants vary in their unique features of morphology, ultrastructure and biochemical properties by species. Furthermore, prey-capturing mechanism as well as structural and physiological adaptations have been used for grouping various carnivorous species. In Drosera plants, glandular trichomes, which develop in the leaf epidermis, are known to play the most important role during the prey capturing process. The present study examined such trichomes, focusing on the glandular type, in leaves of Drosera anglica using scanning and transmission electron microscopy. Three types of rudimentary glandular trichomes were found to develop within the folded leaf primordia and immature leaf during early development. The first type, stalked glandular trichomes (Type I), occurred on the margin and upper epidermis of the leaf. With maturation, the longest glandular trichomes having lengthy stalks, ca. $2.2{\sim}5.1\;mm$, developed along the margin, while shorter stalked trichomes, ca. up to $200\;{\mu}m$, were found on the inner leaf blade. The shorter ones consisted of a globose head having two layers of secretory cells, parenchyma bell cells and tracheids and a multicellular stalk. The stalks gradually decreased in length in centripetal fashion. The second type, Type II, having ca. $15{\sim}30\;{\mu}m$ short stalks, also developed along the inner blade. Both types secreted mucilage from the secretory cells which had a thin cell wall and cuticle layer. The sessile six-celled glandular trichomes were the third type, Type III, and were $25{\sim}40\;{\mu}m$ in length. They were distributed most commonly throughout the upper and lower epidermis, petiole and even on the stalk surfaces of the first two types of trichomes. The third type was also found to be involved in the active secretion. In prey capturing leaves, all trichome types secreted substances through thin cuticles in the head cell wall, which exhibited relatively loose wall components.

Effects of ischemic preconditioning, KATP channel on the SOD activation and apoptosis in ischemic reperfused skeletal muscle of rat (허혈양상화와 KATP 통로가 허혈후 재관류된 흰쥐의 골격근육에서 SOD 활성 및 apoptosis에 미치는 영향)

  • Abn, Dong-choon;Paik, Doo-jin;Yang, Hong-hyun
    • Korean Journal of Veterinary Research
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    • v.39 no.5
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    • pp.878-895
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    • 1999
  • Ischemic preconditioing (IPC), i.e., a preliminary brief episode of ischemia and reperfusion, has been shown to reduce the cell damage induced by long ischemia and reperfusion. Superoxide radical which is produced during reperfusion after ischemia was recognized as a factor of the ischemic injury and it is dismutated into $H_2O_2$ and $O_2$ by two types of intracellular superoxide dismutase (SOD), Cu,Zn-SOD in cytoplasm and Mn-SOD in mitochondria. Recently oxygen free radicals are suggested to induce the apoptosis, however mechanism of the reduced apoptosis by ischemic preconditioing was unknown, while many studies performed in mammalian heart indicated that ATP-sensitive $K^+$ ($K_{APT}$) channel activation related with the protective effects. The aim of present study is to investigate 1) whether IP upregulate the Cu,Zn-SOD and Mn-SOD activities, and 2) whether ischemic preconditioning decreases apoptosis via $K_{APT}$ channel activation in timely reperfused skeletal muscle after long ishemia. The experimental animals, Sprague-Dawley rats weighing 250~300g, were divided into 8 groups; 1) control group, 2) ischemic preconditioning only groups, 3) pinacidil, a $K_{APT}$ channel opener, treatment only groups, 4) glibenclamide, a $K_{APT}$ channel blocker, treatment only groups, 5) ischemia groups, 6) ischemia after IPC groups, 7) ischemia and pinacidil treatment groups, and 8) IP and ischemia after glibenclamide pretreatment groups. Animals of the control group were administered with the vehicle (DMSO) alone. Pinacidil (1mg/kg) was administered intravenously 5 minutes after initiation of ischemia, and glibenclamide (0.5mg/kg) was injected intravenously 20 minutes before IPC. In rats that were ischemic preconditioned, the left common iliac artery was occluded for 5 minutes followed by 5 minutes of reperfusion by three times using vascular clamp. Ischemia was done by occlusion of the same artery for 4 hours. The specimens of left rectus femoris muscle were obtained immediately (0 hour), 12 hours, 24 hours after drug administrations, IP or ischemia and reperfusion. The immunoreactivities of SOD and its alterations were observed by use of sheep antihuman Cu,Zn-SOD and Mn-SOD antibodies on the $10{\mu}m$ cryosections. The incidencies of apoptosis were observed by TUNEL methods with in situ apoptosis detection kit on $6{\mu}m$ paraffine section. The results obtained were as follows : 1. After IPC, immunoreactivities of Cu,Zn-SOD mainly in the small-sized fibers were increased by 24 hours, that of Mn-SOD at 0 hour and 24 hours. 2. No significant changes in immunoreactivities of SOD was observed in the pinacidil and in the glibenclamide treatment only groups, and in the ischemia only groups. 3. The immunoreactivities of the Cu,Zn-SOD were increased in the ischemia after IPC groups and the ischemia and pinacidil treatment groups. 4. The immunoreactivities of the Cu,Zn-SOD in the IPC and ischemia after glibenclamide pretreatment groups were not increased except for the 12 hours reperfusion group. But, Mn-SOD immunoreactivities were increased in the 0 hours, 12 hours and 24 hours after reperfusion. 5. In the control group, the IPC only groups, and the pinacidil treatment only groups, negative or trace apoptotic reactions were observed, but the positive apoptotic reaction occured in the glibenclamide treatment groups. 6. Moderate or many number of apoptosis were revealed in the ischemia groups, and also the IPC and ischemia after glibenclamide pretreatment group except for 12 hours and 24 hours after reperfusion. However, the incidence of apoptosis was decreased in the ischemia after IPC groups and in the ischemia and pinacidil treatment groups. 7. There is a coincidence between the increase of Cu,Zn-SOD immunoreactivities and the decrease of apoptosis in the presence of ischemia and reperfusion. These results suggest that the protective effects of ishemic preconditioing may related to the SOD activation, and the ischemic preconditioning decreases the apoptosis partially via $K_{APT}$ channel activation in timely reperfused rat skeletal muscle. It is also suggested that inhibition of apoptosis by IPC may related with the SOD activation.

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Physical Properties and Apparent Thermal Diffusivity of the Soils where Soil Temperature is Measured Regularly (기상청(氣象廳) 지온(地溫) 측정(測定) 토양(土壤)의 물리적(物理的) 성질(性質)과 겉보기 열확산(熱擴散) 계수(係數) 산정(算定))

  • Song, Kwan-Cheol;Jung, Yeong-Sang;Kim, Byung-Chan;Ahn, Yoon-Soo;Um, Ki-Tae
    • Korean Journal of Soil Science and Fertilizer
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    • v.25 no.3
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    • pp.220-230
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    • 1992
  • Soil temperature is one of the important environmental factors which control all the physical, chemical and biological processes in soil including germination and root growth of plants and other organisms living in the soil ecosystem. Soil water and nutrient availability and mobility are temperature dependent. Soil temperature change is depended primarily upon energy exchange in soil surface, meteorological variance and physical properties of the soils which are closely related to heat transfer mechanism. In this study physical properties including bulk density, soil texture and organic matter content were measured and thermal diffusivity on the soils was calculated. Soil samples from the 66 meteorological stations under the Korea Meteorology were collected and the physical parameters were measured. To obtain relationship between thermal diffusivity and soil water content a heat probe thermal diffusivity measurement apparatus was designed and used in this experiment. According to the survey on soil physicsal properties on the 66 meteorological stations, the 52% of the surface soil texture were sandy loam and laomy sand or sand, 38% were loam and silty loam, and 10% were clay loam and silty clay loam. The bulk density which was closely related with thermal properties showed average of $1.41g/cm^3$ for sandy soils, $1.33g/cm^3$ for loam and silty loam soils, and $1.21g/cm^3$ for clay loam and silty clay loam soils. The apparent thermal diffusivity of the upper layer from 0 to 30cm ranged from 1.16 to $8.40{\times}10^{-3}cm^3/sec$ with average of $3.53{\times}10^{-3}cm^3/sec$. The apparent thermal diffusivities of the Jeju soils of which organic matter contents were high and the bulk densities were low were near $2{\times}10^{-3}cm^3/sec$. The thermal diffusivity of snow measured in Chuncheon ranged from 0.822 to $2.237{\times}10^{-3}cm^3/sec$. The damping depth calculated from the thermal diffusivity ranged from 5.92 to 13.65cm for daily basis and 124 to 342cm for yearly basis. The significant regression equation to estimate thermal diffusivity with bulk density and soil water content was obtained by the heat probe in laboratory.

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Effects of Agrimonia pilosa Ledeb. Water Extract on α-Glucosidase Inhibition and Glucose Uptake in C2C12 Skeletal Muscle Cells (짚신나물 열수 추출물의 α-Glucosidase 저해 효과 및 근육세포에서 포도당 이용에 미치는 영향)

  • Kim, Sang-Mi;Lee, Young Min;Kim, Mi-Ju;Nam, Song-Yee;Kim, Sung-Hee;Jang, Hwan-Hee
    • The Korean Journal of Food And Nutrition
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    • v.26 no.4
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    • pp.806-813
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    • 2013
  • Agrimonia pilosa Ledeb. is a medicinal plant with anti-tumor, anti-oxidant, anti-inflammatory and anti-hyperglycemic activities. However, few studies of the anti-diabetic effect of A. pilosa on insulin resistance status have been performed. In the present study, the anti-diabetic effect of A. pilosa water extract (AP) was determined by investigating its ${\alpha}$-glucosidase inhibitory property, glucose utilization, and uptake, as well as insulin resistance mechanism of action in C2C12 skeletal muscle cells. Compared to positive control (acarbose), AP ($10mg/m{\ell}$) showed a similar ${\alpha}$-glucosidase inhibitory capacity. Glucose uptake was significantly increased by $1{\mu}m$ insulin treatment (p<0.05). However, palmitic acid (FFA, 1 mM) induced muscle insulin resistance and glucose uptake dysfunction. On the other hand, AP ($10{\mu}g/m{\ell}$) was capable of reversing the FFA-induced insulin resistance in C2C12 myotubes. Compared to control, AP ($100{\mu}g/m{\ell}$ without insulin) significantly increased the utilization of glucose (p<0.05) in C2Cl2 myotubes cultured in normal glucose (7 mM). AP treatment significantly increased the relative mRNA and protein expression levels of Akt. In particular, the effect of A. pilosa on the insulin signaling system is associated with the up-regulation of Akt genes and glucose uptake in C2Cl2 myotubes. These results suggest that A. pilosa is useful in the prevention of diabetes and the treatment of hyperglycemic disorders.

The Role of Oxygen Free Radicals from Endothelial Cells in Endotoxin-induced Endothelial Cell Cytotoxity (내독소에 의한 혈관 내피세포 손상에서 혈관 내피세포로부터 유리된 산소기의 역할에 관한 연구)

  • Choi, Hyung-Seok;Jeong, Ki-Ho;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol;Jung, Ki-Suck
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.4
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    • pp.319-327
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    • 1994
  • Background: The pathogenetic mechanism of adult respiratory distress syndrome(ARDS) is not clearly defined yet, but it is well known that increased pulmonary capillary permeabilty is characteristic feature of ARDS. The increased alveolar-capillary permeability is usually preceded by damage of pulmonary artery endothelial cells. The released enzymes and oxygen free radicals from the activated neutrophils seem to play a predominant role in endothelial cell cytotoxicity. The activated neutrophils, however, probably are not the sole contributing factor in this type of damage because many cases of ARDS have been reported in severe neutropenia. Bacterial endotoxin perse and/or oxygen free radicals released from endothelial cells are suggested to be possible factors that contribute to the development of ARDS. The purpose of this study is to investigate the direct cytotoxicity of endotoxin and the role of oxygen free radicals released from the endothelial cells in endotoxin-induced endothelial cell cytotoxicity. Methods: First, to investigate whether endotoxin is cytotoxic to HUVE by itself, various doses of endotoxin were added to culture medium and cytotoxicity was measured. Second, to evaluate the possible role of oxygen free radical in endotoxin-induced HUVE cytotoxicity, various antioxidants were added on the endotoxin-induced HUVE cytotoxicity and cytotoxicity was measured. Third, to verify the release of oxygen free radicals from HUVE, the concentrations of hydrogen peroxide in the endotoxin-treated culture supernatant were measured. Finally, to observe the cytotoxic effect of hydrogen peroxide, HUVE cytotoxicity in the presence of various doses of hydrogen peroxide was measured. The fourth generations of subcultured HUVE from primary culture were used. The cell cytotoxicity was quantified by the chromium-51 release assay. Results: 1) Endotoxin alone showed HUVE cytotoxicity in a dose-dependent fashion. 2) Endotoxin-induced HUVE cytotoxicity was significantly attenuated by the pretreatment of catalase and DMTU. 3) Hydrogen peroxide was released from HUVE after endotoxin treatment in a dose-dependent fashion. 4) Exogenous hydrogen peroxide also showed HUVE cytotoxicity in a dose-dependent fashion. Conclusion: These results suggest that endotoxin alone can directly injure HUVE, and, oxygen-free radicals released from HUVE in response to endotoxin may also participate in the endotoxin-induced HUVE cytotoxicity.

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