Lipopolysaccharide 세포독성에 대한 SNP 효과에 관한 연구

황호근,허정무,방현주,김범영,손홍문,문연자,오재민,정연태,최민규 대한해부학회 2000 Anatomy & Cell Biology Vol.33 No.5

Effect of Lipopolysaccharide (LPS) Exposure on the Reproductive Organs of Immature Female Rats

Da Kyung Yoo,Sung-Ho Lee 한국발생생물학회 2016 발생과 생식 Vol.20 No.2

Lipopolysaccharide (LPS), an endotoxin, elicits strong immune responses in mammals. Several lines of evidence demonstrate that LPS challenge profoundly affects female reproductive function. For example, LPS exposure affects steroidogenesis and folliculogenesis, resulting in delayed puberty onset. The present study was conducted to clarify the mechanism underlying the adverse effect of LPS on the delayed puberty in female rats. LPS was daily injected for 5 days (50 μg/kg, PND 25-29) to treated animals and the date at VO was evaluated through daily visual examination. At PND 39, animals were sacrificed, and the tissues were immediately removed and weighed. Among the reproductive organs, the weights of the ovaries and oviduct from LPS-treated animals were significantly lower than those of control animals. There were no changes in the weights of uterus and vagina between the LPS-treated and their control animals. Immunological challenge by LPS delayed VO. Multiple corpora lutea were found in the control ovaries, indicating ovulations were occurred. However, none of corpus luteum was present in the LPS-treated ovary. The transcription level of steroidogenic acute regulatory protein (StAR), CYP11A1, CYP17A1 and CYP19 were significantly increased by LPS treatment. On the other hand, the levels of 3β-HSD, 17β-HSD and LH receptor were not changed by LPS challenge. In conclusion, the present study demonstrated that the repeated LPS exposure during the prepubertal period could induce multiple alterations in the steroidogenic machinery in ovary, and in turn, delayed puberty onset. The prepubertal LPS challenge model used in our study is useful to understand the reciprocal regulation of immune (stress) - reproductive function in early life.

大防風湯의 LPS誘發 關節炎 緩和效果에 관한 實驗的 硏究

田元準,辛吉祚,李源哲 동국대학교 한의학연구소 2000 東國韓醫學硏究所論文集 Vol.9 No.-

본 실험은 大防風湯이 혈관생성과 세포유착인자발현 억제를 통해 LPS(lipopolysaccharide)유발 관절염 완화에 미치는 영향을 조사하기 위해 LPS주사로 관절염이 유발된 생쥐에게 大防風湯 추출액을 음용 투여하였다. 실험동물은 20g된 female Balb/c계 생쥐암컷을 선별하여 아무런 처치를 하지 않은 대조군, 무릎관절에 LPS를 주사한 LPS군, LPS주사후 大防風湯 추출액을 투여한 DBP군으로 나누어, 다시 LPS와 DBP군을 LPS처리 후 시간의 경과에 따라 3, 7, 14일로 구분하였으며, 실험군 각각에는 실험동물을 10마리씩 배정하였다. 광학 현미경(BX50)으로 大防風湯 추출액 투여가 LPS주사 후 증가된 윤할 관절막내 혈관분포 증가에 미치는 영향과 아울러 윤활세포, 섬유화 및 섬유모세포, 비만세포와 염증관여세포, 세포유착인자 등의 윤활 관절막에서의 형태변화를 관찰하였다. DBP군에서는 자유면에서 윤활세포과형성의 억제와 침윤 림프구의 감소, 낮은 섬유화 양상과 침적 섬유모세포의 감소, 모세혈관 주변의 림프구 침적 감소와 비만세포 분포감소, 자유면과 섬유지지역분포 ICAM과 혈관주변분포VCAM 등과 같은 세포유착인자의 감소 등이 관찰되었다. 이상의 결과로 볼 때 大防風湯은 관절염 유발시 발생되는 혈관생성과 세포유착인자의 발현증가 등을 억제함으로써 이후 발생되는 윤활 관절막 손상을 완화하는 것으로 사료된다. Purpose : This study is to investigate the alleviation effects of Daebangpoongtang in LPS induced arthritis in mice knee joint. Methods : Daebangpoongtang was chosen to treat the arthritis caused by injecting 300 μg/kg LPS to mice knee joint. The control group had no treatment, while the LPS group was injected 300μg/kg LPS to mice knee joint and the DBP group was oral administrated of Daebangpoongtang. After injection of 300μg/kg LPS to mice knee joint, the alteration of synovial lining cell, vessel, fibrosis, distribution of collagen fiber, fibroblast, mast cell, infiltration of inflammation component cell and distribution of ICAM and VCAM was observed by light microscope(BX50). Results : In the DBP extract treatment group, the distribution of vessel, the enlargement of synovial lining cell layer, the synovial lining cells with filopodia, the fibrosis, the distribution of fibroblast in synovial membrane, the distribution of ICAM and VCAM on the knee joint was less than that of LPS group. Infiltrated lymphocyte into the apical surface had not observed in the DBP extract treatment group. The distribution of mast cell was as same as control group(no treatment group) and it showed granulated type. Conclusion : According to the above results, it might be considered that the administration of Daebangpoongtang has a curative effect on synovial membrane injury in arthritis by inhibiting increase of vessel, cell adhesion molecule(1CAM and VCAM) in LPS induced arthritis.

금은화가 LPS로 유발된 급성 폐 손상에 미치는 영향

이창건,최해윤,박미연,김종대,Yi, Chang-Geon,Choi, Hae-Yun,Park, Mee-Yeon,Kim, Jong-Dae 대한예방한의학회 2011 대한예방한의학회지 Vol.15 No.1

Objective : The object of this study was to observe the effects of Lonicerae Flos (LF) aqueous extracts on lipopolysaccharide (LPS)-induced rat acute lung injury. Method : Five different dosages of LF extracts were orally administered once a day for 28 days before LPS treatments, and then all rats were sacrificed after 5 hour-treatment of LPS. Eight groups of 16 rats each were used in the present study. The following parameters caused by LPS treatment were observed ; body weights, lung weights, pulmonary transcapillary albumin transit, arterial gas parameters (pH, $PaO_2$ and $PaCO_2$) bronchoalveolar lavage fluid (BALF) protein lactate dehydrogenase (LDH), and proinflammatory cytokines tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$) contents, total cell numbers, neutrophil and alveolar macrophage ratios, lung malondialdehyde (MDA), myeloperoxidase (MPO), proinflammatory cytokines TNF-${\alpha}$ and IL-$1{\beta}$ contents. In addition, the histopathologic changes were observed in the lung in terms of luminal surface of alveolus, thickness of alveolar septum, number of polymorphonuclear neutrophils. Result : As results of LPS-injection, dramatical increases in lung weights, pulmonary transcapillary albumin transit increases, increases in $PaCO_2$, decreases in pH of arterial blood and $PaO_2$, increases of BALF protein, LDH, TNF-${\alpha}$ and IL-$1{\beta}$ contents, total cells, neutrophil and alveolar macrophage ratios, TNF-${\alpha}$ and IL-$1{\beta}$ contents increases were detected with decreases in LSA and increases of alveolar septum and PMNs numbers, respectively as compared with intact control. These are means that acute lung injuries (resembling acute respiratory distress syndrome) are induced by treatment of LPS mediated by inflammatory responses, oxidative stress and related lipid peroxidation in the present study. However, these LPS-induced acute lung injuries were inhibited by 28 days continuous pretreatment of 250 and 500mg/kg of LF extracts. Because of lower three dosages of LF treated groups, 31.25 and 62.5 and 125mg/kg did not showed any favorable effects as compared with LPS control, the effective dosages of LF in LPS-induced acute lung injuries in the present study, is considered as about 125mg/kg. The effects of 250mg/kg of LF extracts showed almost similar effects with ${\alpha}$-lipoic acid 60mg/kg in preventing LPS-induced acute lung injuries. Conclusion : It seems that LF play a role in protecting the acute respiratory distress syndrome caused by LPS.

Sulfolaphane이 lipopolysaccharide (LPS)에 의해 유도된 matrix metalloproteinase-9(MMP-9) 발현에 미치는 영향

Jung Tae Lee(이정태),Kyung Jin Woo(우경진),Taeg Kyu Kwon(권택규) 한국생명과학회 2010 생명과학회지 Vol.20 No.2

Sulforaphane은 십자가화 채소에 존재하는 화합물로 항염증, 항암 및 신생혈관 생성의 억제 효과가 알려짐으로써 최근 많은 연구가 활발히 이루어지고 있으나, LPS에 의한 MMP-9 활성 조절에 대한 연구는 매우 미흡한 편이다. 따라서 본 연구에서 sulforaphane이 LPS 유도에 의한 MMP-9 활성에 미치는 영향에 대해서 조사해 보았다. Raw 264.7 세포에 sulforaphane을 전처리 한 후 LPS를 처리하여 gelatin zymography를 실시해 본 결과, LPS에 의해 유도된 MMP-9 활성 증가가 sulforaphane 농도 의존적으로 감소됨을 확인 하였다. 또한 RT-PCR과 MMP-9의 luciferase assay를 통한 실험에서 sulforaphane의 MMP-9 억제효과가 전사단계에서 조절됨을 추측 할 수 있었다. MMP-9 promoter 부위에 여러 가지의 전사조절인자 결합부위가 존재한다. 특히 AP-1과 NF-κB가 중요 전사조절인자로 작용하여 MMP-9 발현조절에 관여한다. 본 실험에서 sulforaphane에 의한 MMP-9 억제효과 기전에 이들 전사조절인자들의 중요한 역할을 조사하였다. AP-1과 NF-κB 결합부위를 변형 시킨 vector를 transfection하여 MMP-9의 promoter 활성을 측정한 결과, 정상 vector에 비해 그 활성도가 현저히 떨어짐을 확인하였고, LPS에 의해 증가되는 AP-1과 NF-κB의 basal promoter 활성 또한 sulforaphane에 의해 감소됨을 관찰 할 수 있었다. 이상의 결과에서 sulforaphane의 MMP-9 활성억제효과는 AP-1과 NF-κB와 같은 전사인자들이 MMP-9의 전사를 조절함으로써 일어나는 것임을 알 수 있었다. 그리고 sulforaphane은 세포의 invasion능력 또한 효과적으로 억제시킴을 관찰 할 수 있었는데 이는 MMP-9 활성억제효과와 밀접한 관련이 있음을 추측 할 수 있었다. Sulforaphane is a naturally occurring member of the iosothiocyanate family, which reveals chemopreventive capacities including anti-cancer, anti-inflammation and inhibition of MMP-9 activities. In this study, we investigated the effect of sulforaphane on the expression of matrix metalloproteinase-9 (MMP-9) in lipopolysaccharide (LPS)-induced Raw 264.7 cells. Sulforaphane strikingly suppressed the LPS-induced MMP-9 activity and mRNA expression in a dose-dependent manner. In addition, sulforaphane inhibited not only the LPS-induced MMP-9 promoter activity but also LPS-mediated activator protein-1 (AP-1) and nuclear factor-κB (NF-κB) promoter activity. Transient transfection by MMP-9 constructs, in which specific transcriptional factors were mutagenized, indicated that the effects of LPS and sulforaphane were mediated via AP-1 and NF-κB response elements. We found that sulforaphane had the ability to suppress LPS-induced invasion in vitro. Taken together, these results demonstrated that sulforaphane effectively suppressed LPS-induced MMP-9 expression via modulation of promoter elements (AP-1 and NF-κB) in MMP-9 transcriptional activation.

5‑Methoxytryptophan pretreatment alleviates lipopolysaccharide‑induced cardiac injury and dysfunction

Fu Yang,Dong Yi-Fei 한국응용생명화학회 2022 Applied Biological Chemistry (Appl Biol Chem) Vol.65 No.3

Reducing inflammation is a promising therapeutic approach for sepsis-induced cardiomyopathy (SIC). The 5-Methoxytryptophan (5-MTP) is a tryptophan metabolite that demonstrates anti-inflammatory, anti-fibrosis, anti-tumorigenesis, and anti-senescence features. Current investigations aimed to assess the 5-MTP pretreatment impacts on lipopolysaccharide (LPS)-induced cardiac injury and dysfunction. For in vivo studies, the mice were categorized randomly into four groups: control, LPS, LPS+5-MTP (25 mg/kg) and LPS+5-MTP (50 mg/kg). The mice in the LPS+5-MTP groups were given 5-MTP intraperitoneally once a day for 7 days. LPS (10 mg/kg) was then administered intraperitoneally for 24 h. Echocardiography, cardiac injury biomarkers, and H & E staining evaluated heart anatomy and function. The findings indicate that 5-MTP pretreatment significantly reduced LPS-induced heart dysfunction and morphological alterations. Western blot assay was used for investigating molecular mechanisms. After LPS stimulation, the pro-inflammatory cytokines (IL-1β, IL-6, TNF-α and NLRP3) protein levels increased while anti-inflammatory cytokine (IL-10) decreased; however, 5-MTP pretreatment mitigated this response by suppressing the stimulation of the NF-κB signaling pathway. Furthermore, 5-MTP administration reduced LPS-induced cardiac apoptosis, as demonstrated by increased protein levels of cleaved-Casepase-1, cleaved-Casepase-3 and Bax, and decreased protein level of Bcl-2 after LPS stimulation, whereas LPS-induced cardiac apoptosis was reversed by 5-MTP pretreatment. In vitro, 5-MTP pretreatment had a similar cardioprotective effect on cultured cardiac fibroblasts challenged with LPS. In conclusion, 5-MTP pretreatment can reduce LPS-induced cardiac inflammation and apoptosis, implying that 5-MTP is a possible therapeutic option for SIC.

젖소에 있어 아급성제1위과산증(SARA)의 강도가 혈장 Lipopolysaccharide (LPS)의 농도에 미치는 효과

손혜인,백성광,문주연,안의영,이현준,손용석,Son, Hey In,Baek, Seong Gwang,Moon, Ju Yeon,Ahn, Eui-Young,Lee, Hyun-Jun,Son, Yong Suk 한국초지조사료학회 2013 한국초지조사료학회지 Vol.33 No.4

본 시험은 젖소 4두에 대하여 제1위체류형 센서(Indwelling probe)를 정치시키고 이를 이용하여 아급성제1위과산증(SARA)의 강도를 조사함과 동시에 pH가 혈 중 내독소(Lipopolysaccharide; LPS)의 농도에 미치는 효과를 알아보기 위하여 수행되었다. 비섬유성탄수화물(Non-fiber carbohydrates, NFC)을 기준으로 적정 수준의 NFC를 포함하고 있는 정상 사료군(Normal diet, ND)과, 아급성제1위과산증(Subacute ruminal acidosis, SARA) 유발가능 사료군(Acidogenic diet, AD)의 2개 처리구에 각 2두씩 배치하여 Cross-over 설계로 사양실험을 반복 실시하였다. 제1위체류형 센서는 장시간에 걸친 제1위 내 pH의 변화를 추적하는 데 효과적이었다. 얻어진 결과에서 AD 급여는 반추위내용물의 pH를 낮추는 데 커다란 영향을 주었으며, 반추위 내 pH의 저하는 혈 중 LPS의 농도가 증가로 이어짐을 확인할 수 있었다. 따라서 혈 중 LPS 농도는 급성 또는 만성형을 불문하고 과산증을 확인하는 지표로 사용 가능할 것으로 판단된다. 또한 착유우용 사료첨가물로 많이 이용되는 소디움벤토나이트(NaB)와 프로필렌글리콜(PG)의 급여는 반추위 내 pH의 저하를 완충시켜 궁극적으로 SARA의 강도를 완화시키고 혈 중 Lipopolysaccharides (LPS) 농도의 증가를 줄여 줌으로써 내독소에 의해 발생하기 쉬운 제엽염 등의 부작용을 완화시키는 데 도움을 줄 수 있을 것으로 사료된다. A study was conducted to determine if subacute ruminal acidosis (SARA) induced by feeding high level of non-structural carbohydrates results in increases in lipopolysaccharides (LPS) of peripheral blood in dairy cattle. In this experiment four Holstein steers, two of which having ruminal canulae with indwelling probes placed for measuring long term pH changes, were alloted into two dietary treatments in a cross-over design, where an acidogenic diet (AD) was formulated by including high amount of non-structural carbohydrates (NSC) based on corn silage and corn flake as TMR ingredients. Data for ruminal pH change and plasma LPS concentration were compared against normal diet (ND) which contained grass hay as forage and low NSC ingredients. Feeding AD for more than 14 days to animals brought about a pH change as low as less than 5.8 for more than 4 hours, which made good contrast to ND fed animals. Decreased ruminal pH also had an effect on LPS concentrations which showed significantly higher level for AD compared with ND. Therefore, plasma LPS concentration may be used as an effective indicator to verify acidosis whether it is acute or chronic. Na-bentonite and Propylene glycol, which are frequently included in dairy TMR as additives, helped decrease ruminal pH by buffering and then ultimately alleviates SARA. Therefore, it could also be helpful to lower the occurrence of laminitis which is often caused by increased blood endotoxin (LPS) concentration.

Lipopolysaccharide, Dexamethasone, 및 N-Nitro-L-Arginine Methyl Ester가 흰쥐 간 조직의 프리라디칼 발생과 제거에 미치는 영향

한기정,김석배,김양균,윤혜영,백광진,이희성,권년수 중앙대학교 의과대학 의과학연구소 2003 中央醫大誌 Vol.28 No.1

Lipopolysaccharide (LPS) stimulates various immune and inflammatory reactions to induces the generation of reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI). ROI and RNI are free radicals with very high reactivity, oxidize cellular constituents including proteins and lipid membranes, and thus play important roles in the pathogenesis of tissue damages in LPS-induced septic shock. The present study examined effects of dexamethasone (DexM), and N-nitro-L-arginine methyl ester (L-NAME) on the generation of ROI and RNI, and on the activities of enzymes removing ROI in the liver of rat model of septic shock. DexM has widely been used as an immunosuppressant. L-NAME has known to block synthesis of nitric oxide (NO), the main RNI, by inhibiting NO synthase (NOS). LPS (10 mg/kg body weight) markedly enhanced NO generation in serum and liver. The increased NO generation was completely blocked by pretreatment with DexM (10 mg/kg) and effectively inhibited by L-NAME (10 mg/kg). Induction of NOS protein expression by LPS was demonstrated by a Western analysis. The NOS expression was completely blocked by DexM pre-treatment, and markedly inhibited by L-NAME. However, the generation of superoxide radical, an important ROI, was not enhance in rat liver by LPS, rather it was decreased a little. Superoxide radical production was increased by L-NAME treatment or combined treatment of DexM and LPS. The level of lipid peroxidation, an index for free radical-induced cell damage, was increased by either LPS or DexM. The enhancement of lipid peroxidation by DexM was abolished by adding LPS. DexM markedly enhanced the activities of ROI removing enzymes, superoxide dismutase, glutathione peroxidase and catalase. However, rats were treated with LPS in addition to DexM, the elevated activities of superoxide dismutase, glutathione peroxidase and catalase were abolished. These results provide information on the generation and removal of ROI and RNI by LPS, DexM and L-NAME in rat liver, and on development of drugs to inhibit free radical-induced liver damage in sepsis.

Autodisplay of the La/SSB protein on LPS-free E. coli for the diagnosis of Sjogren's syndrome

Yoo, G.,Dilkaute, C.,Bong, J.H.,Song, H.W.,Lee, M.,Kang, M.J.,Jose, J.,Pyun, J.C. IPC Science and Technology Press ; Elsevier Scienc 2017 Enzyme and microbial technology Vol.100 No.-

<P>The objective of this study was to present an immunoassay for the diaghosis of Sjogren's syndrome based on the auto displayed La/SSB protein on the outer membrane of intact E. coil (strain UT-5600) and LPS-free E. coil (ClearColi (TM)). As the first step, an autodisplay vector (pCK002)was transfected into intact E. coli and LPS-free E. coli for comparison of efficiency of autdisplay of La/SSB. The Maximal level of La/SSB expression was estimated to be similar for LPS-free E. coli and intact E. coli at different optimal induction periods. Intact E. coli was found to grow twofold faster than LPS-free E. coli, and the maximal level of expression for LPS-free E. coil was obtained with a longer induction period. When the zeta potential was measured, both intact E. coli and LPS-free E. coli showed negative values, and the autodisplay of negatively charged La/SSB protein (pI < 7) on the outer membrane of intact E. coil and LPS-free E. coli resulted in a slight change in zeta potential values. E. coli with autodisplayed La/SSB protein was used for an immunoassay of anti-La/SSB antibodies for the diagnosis of Sjogren's syndrome. The surface of E. coli with the autodisplayed antigen was modified with rabbit serum and papain to prevent false positive signals because of nonspecific binding of unrelated antibodies from human serum. LPS-free E. coli With autodisplayed La/SSB protein yielded sensitivity and selectivity of 81.6% and 78.6%, respectively. The Bland-Altman test showed that the immunoassays based on LPS-free E. coli and intact E. coli with autodisplayed La/SSB protein were statistically equivalent to a clinical immunoassay for detection of anti-La/SSB antibodies (confidence coefficient 95%). (C) 2017 Elsevier Inc. All rights reserved.</P>

Lipopolysaccharide Inhibits Proliferation of the Cultured Vascular Smooth Muscle Cells by Stimulating Inducible Nitric Oxide Synthase and Subsequent Activation of Guanylate Cyclase

Hyoung Chul Choi,Sang Gon Lee,Jong Ho Kim,Joo Young Kim,Uy Dong Sohn,Jeoung Hee Ha,Kwang Youn Lee,Won Joon Kim 대한생리학회-대한약리학회 2001 The Korean Journal of Physiology & Pharmacology Vol.5 No.4

<P> This study was undertaken to investigate the mechanism of lipopolysaccharide (LPS) and nitric oxide (NO) as a regulator of vascular smooth muscle cell (VSMC) proliferation. VSMC was primarily cultured from rat aorta and confirmed by the immunocytochemistry with anti-smooth muscle myosin antibody. The number of viable VSMCs were counted, and lactate dehydrogenase (LDH) activity was measured to assess the degree of cell death. Concentrations of nitrite in the culture medium were measured as an indicator of NO production. LPS was introduced into the medium to induce the inducible nitric oxide synthase (iNOS) in VSMC, and Western blot for iNOS protein and RT-PCR for iNOS mRNA were performed to confirm the presence of iNOS. Inhibitors of iNOS and soluble guanylate cyclase (sGC), sodium nitroprusside (SNP) and L-arginine were employed to observe the action of LPS on the iNOS-NO-cGMP signalling pathway. LPS and SNP decreased number of VSMCs and increased the nitrite concentration in the culture medium, but there was no significant change in LDH activity. A cell permeable cGMP derivative, 8-Bromo-cGMP, decreased the number of VSMCs with no significant change in LDH activity. L-arginine, an NO substrate, alone tended to reduce cell count without affecting nitrite concentration or LDH level. Aminoguanidine, an iNOS specific inhibitor, inhibited LPS-induced reduction of cell numbers and reduced the nitrite concentration in the culture medium. LY 83583, a guanylate cyclase inhibitor, suppressed the inhibitory actions of LPS and SNP on VSMC proliferation. LPS increased amounts of iNOS protein and iNOS mRNA in a concentration-dependent manner. These results suggest that LPS inhibits the VSMC proliferation via production of NO by inducing iNOS gene expression. The cGMP which is produced by subsequent activation of guanylate cyclase would be a major mediator in the inhibitory action of iNOS-NO signalling on VSMC proliferation.