The elderly suffer from an impaired immune function being obvious in a higher susceptibility to infections. Although the inflammatory cells are the major immunomodulatory cells, fibroblasts also secrete a variety of inflammatory cytokines and chemokines. Therefore periodontal tissue aging might playa role in development and progress of periodontitis. In this study, we investigated the effect of in vitro periodontal ligament cellular aging on the inflammatory cytokines, chemokines, and matrix metalloprotease(MMP)-2 expression induced by lipopolysaccharide(LPS) treatment. Three different cell populations were used; passages 4-5, 14-15, and 24-25 (at passage 27, more than 90% cells were replicative senescent). LPS increased the expression of interleukin(IL)-1β, IL-6, and tumor necrosis factor-α, IL-8, RANTES, and MMP-2. However, the order of induction folds were passages 14-15 > 4-5 > 24-25. While the expression level of Toll-like receptor(TLR) 4 decreased according to the increase in passage number, the level of TLR2 was highest at passages 14-15 and then decreased at passages 24-25. While the spontaneous expression of IL-8 decreased according to the increase in passage number, that of RANTES and proMMP-2 increased according to the increase in passage number. These results suggest that the aging of periodontal ligament fibroblasts differentially affect the role as immunomodulatory cells in response to periodontopathic bacteria and therefore might be another risk factor of periodontitis progression.

With a rabbit model, the present study was performed to examine the effect of Escherichia coli lipopolysaccharide (E. coli LPS) on hormones. Cortisol, epinephrine, and norepinephrine concentrations in LPS-treated groups were high at all sampling periods (from 3 hrs to 72 hrs) as compared to control group (p$lt;0.05 or p$lt;0.01). The each peak time was at respectively 24 hrs, 3 hrs, and 12 hrs. Insulin and glucagon concentrations in LPS-treated groups elevated up to 12 hrs (p$lt;0.01 or p$lt;0.05) with a each peak point at 12 hrs or 6 hrs, while those of the rest sampling points (from 24 hrs to 72 hrs) were lower than that of control (p$lt;0.05). Increase of cortisol concentration was generally dose-dependent, whereas the changes of the other hormones were irregular patterns. These observations show that E. coli LPS lead to releases of stress hormones such as cortisol, epinephrine, and norepinephrine and disturbances of endocrine systems. These LPS-induced hormonal disorders may cause physiologically fatal results.

근류균과 두과작물의 공생에서 숙주결합성을 조사하기 위하여 대두 종자 및 및 유근으로부터 분리한 lectin과 근류균의 표피다당과의 결합성을 확인한 결과는 다음과 같다. 팔달, 백운 및 황금으로부터 분리한 종자선을 형성하였다. 대두 종자 lectin 및 뿌리추출물은 R. japonicum및 B. japonicum과 결합하였고 R. viceae와는 결합 하지 않았으며 완두의 lectin은 R. viceae와 결합하였으나 R. japonicum및 B. japonicum과는 결합 하지 않았다. B. japonicum과 R. viceae로 부터 분리한 세포표피 다당의 겔 여과한 결과 두개의 분획으로 각각 나타났다. B. japonicum으로 부터 분리한 표피다당은 대두의 종자 lectin과 결합하였으나 R. viceae로 부터 분리한 표피다당은 대두 lectin과 결합하지 않았다. 이상의 결과로 부터 근류균이 숙주세포와 결합할 때 숙주를 인식하는 초기 단계에서 lectin이 관여함을 추론할 수 있고 이것이 상호접종군을 형성하는데 매개역할을 하는 것으로 추정할 수 있었다.

It is study was designed to characterize endotoxin-induced prostaglandin production in primary cultured rat vascular smooth muscle cells (VSMC). The time course for prostag)andim synthesis in lipopolysaccharide (LPS)-stimulated VSMC showed that the maximum production was reached in 12 hours. LPS induced prostaglandin H2 synthase (PGHS) activity in VSMC and the time course profile in the changes of PGHS activity paralleled that of total prostaglandin production. Differential treatment showed that 4 hours' exposure to LPS was enough for the maximum effect on the prostaglandin production and this effect was completely inhibited by the co-treatment of actinomycin D, a transcription inhibitor. These results suggest that LPS effect might be determined within 4 hours. Actinomycin D increased PGHS activity without affecting prostaglandin production if added 4 hours after LPS treatment. On the other hand, cycloheximide, a translation inhibitor, augmented LPS-induced prostaglandin production if treated during first four hours, but it inhibited LPS-induced PGHS activity regardless of treatment schedule. These results suggeat the existence of multiple regulating mechanisms in the LPS-induced prostaglandin synthesis.

Vibrio vulnificus에서 lipopolysaccharide(LPS)를 추출하여 지방산 조성을 분석한 후 이 결과를 Escherichia coli LPS와 Salmonella typhimurium LPS의 것들과 비교하였다. Vibrio vulnificus LPS의 주 지방산은 myristic acid(C14:0, 41.37%)이었고 Escherichia coli LPS는 lauric acid(C12:0, 37.03%), Salmonella typhimurium LPS는 capric acid(C10:0, 48.60%)로 서로 달랐으나 이 세가지 지방산이 각 LPS의 주성분이었다(70%이상).

Background : In ancient, roots of Rumex crispus, called wooi-daehwang, were used for various symptoms and diseases like cough, phlegm, bronchitis and hepatitis, caused inflammatory. As a part of ongoing research to elucidate and characterize anti-inflammatory nutraceuticals, solvent-partitioned fractions from R. crispus root were tested for their ability to suppress inflammation. In this study, NO synthesis inhibitory activity of solvent-partitioned fractions from R. crispus root on LPS-stimulated RAW264.7 mouse macrophages was evaluated. Methods and Results : The EtOH extracts were suspended in water. The aqueous layer was further partitioned in diethylether, ethylacetate and n-butanol, sequentially. RAW264.7 cells were seeded onto 96-well plates, and cells were allowed to adhere for 6 h and then were pretreated with the R. crispus root extracts for 24 h. Cellular nitric oxide (NO) production was stimulated by adding lipopolysaccharide (LPS). Absorbance was measured at 520 ㎚ by microplate reader. NO synthesis inhibitory activity potential of these fractions was evaluated by assessing NO production by LPS-stimulated RAW264.7 cells in the presence and absence of the solvent-partitioned R. crispus root fractions. NO synthesis inhibitory activity of diethylether fraction diluted in 50 ㎍/㎖, 25 ㎍/㎖, 12.5 ㎍/㎖, 6.25 ㎍/㎖, 3.125 ㎍/㎖ was 79.2%, 70.9%, 59.5%, 16.1%, and 11.8%, respectively. And NO synthesis inhibitory activity range of another fractions, EtOAc, n-BuOH and aqueous layer, were 0 - 30.2%, 0 - 20.1% and 3.8 - 22.4%, respectively. Conclusion : From the above results, it showed that diethylether fraction have strong NO synthesis inhibitory activity, it was suggested that R. crispus root have NO synthesis inhibitory effects. R. crispus root possesses anthraquinones, such as chrysophanol, parietin, and anthrones etc. According to previous studies, R. crispus semen extract has analgesic and hepatoprotective effect as anti-inflammatory, and extract of R. napalensis has cyclooxygenase (COX)-2, COX-1 inhibitory and free radical scavenging effect. Our present study has shown that R. crispus root extracts anti-inflammatory effects probably by suppressing iNOS expressions, and resulting in the inhibition of NO synthesis.

본 연구의 목적은 토종다래의 용매별 추출물에 따른 약리활성에 대한 검증 및 효능 평가로서 토종다래의 항산화, 항염증에 대한 효과를 확인하였다. 염증 반응은 자극이 가해지면 histamine, serotonin, prostaglandin과 같은 혈관 활성물질에 의해 혈관 투과성이 증대되어 염증을 유발하고 cytokine, free radical, lysosomal enzyme 등 다양한 매개 인자가 관여한다. 자극에 의한 macrophage cell의 염증반응은 tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), interleukin-1β(IL-1β)와 같은 pro-inflammatory cytokine의 발현이 유도되고, inducible nitric oxide synthase(i-NOS)와cyclooxygenase-2(COX-2)에 영향을 받는 유전자의 발현을 자극하게 되어 nitric oxide(NO) 등의 염증 인자가 생성된다. 이에 따라 토종다래 추출물의 항염증에 대한 연구를 위해 이에 영향을 주는 인자인 i-NOS, COX-2의 단백질 발현억제 작용을 확인하였다. 그 결과, HKE > HKA > HKW 순서로 높은 효능을 확인 할 수 있었다. 가장 효과가 좋은 HKE 처리군에서 다양한 염증성 인자의 mRNA 발현량을 확인하였다. 측정 결과, HKE(2,000 μg/mL)는 i–NOS, COX-2, IL-1β, IL-6, TNF-α mRNA 발현이 각각 93.2%, 27.9%, 96.4%, 89.4%, 73.9% 억제되는 효과를 확인할 수 있었다. 또한, HKE의 nuclear factor kappa B(NF-κB) 단백질 발현에 농도의존적으로 유의미한 결과를 확인하였으며, 이에 토종다래의 항염증효과는 LPS에 의한 TLR4의 자극에서 NF-κB 경로의 완화로 나타는 것임을 검증하였다. 결론적으로 토종다래는 70% ethanol 추출물(HKE)의 항염증 효과가 가장 높았으며, HKE는 대식세포에서 NF-κB 염증관련 경로의 억제로 세포 내 mRNA 및 단백질 수준에서의 염증인자들의 생성을 저해하여 항염증 효과가 명백히 확인되었다. 향후 본 연구팀은 토종다래의 항염증과 관련된 유효성분의 분리정제 및 구조분석을 진행할 예정이다.