Smilax guianensis Vitman Extract Prevents LPS-Induced Inflammation by Inhibiting the NF-κB Pathway in RAW 264.7 Cells

김주경,김민정,이지수,Kongmany Sydara,이상우,변상균,정성근 한국미생물·생명공학회 2020 Journal of microbiology and biotechnology Vol.30 No.6

Nutraceutical treatments can reduce inflammation and prevent the development of inflammatory diseases. In this study, the anti-inflammatory effects of Smilax guianensis Vitman extract (SGE) were examined. SGE suppressed lipopolysaccharide (LPS)-mediated nitrite production in RAW 264.7 cells. SGE also prevented the LPS-induced expression of inducible nitric oxide synthase (iNOS) but not cyclooxygenase (COX)-2. Western blot analysis showed that SGE attenuated LPS-induced phosphorylation of IκB kinase (IKK), inhibitor of kappa B (IκB), and p65. Additionally, SGE inhibited LPS-induced IκB degradation in RAW 264.7 cells. Western blot analysis of the cytosolic and nuclear fractions, as well as immunofluorescence assay results, revealed that SGE suppressed LPS-induced p65 nuclear translocation in RAW 264.7 cells. Moreover, SGE reduced LPS-induced interleukin (IL)- 1β, IL-6, and tumor necrosis factor-α (TNF-α) mRNA expression and IL-1β and IL-6 protein expression in RAW 264.7 cells. Collectively, these results indicate that SGE suppresses the NF-κB signaling pathway and thereby inhibits the production of NO, IL-1β, and IL-6.

Croton hirtus L'Hér Extract Prevents Inflammation in RAW264.7 Macrophages Via Inhibition of NF-κB Signaling Pathway

김민정,김주경,Kongmany Sydara,이상우,정성근 한국미생물·생명공학회 2020 Journal of microbiology and biotechnology Vol.30 No.4

Consumption of anti-inflammatory nutraceuticals may help treat or prevent inflammation-related illnesses such as diabetes, cardiovascular disease, and cancer. This study evaluated the effect of Croton hirtus L'Hér extract (CHE) on lipopolysaccharide (LPS)-induced nitric oxide (NO) production and nuclear factor kappa-B (NF-κB) signaling cascades. CHE significantly suppressed LPS-induced NO production and inducible nitric oxide synthase (iNOS) expression in RAW264.7 macrophages, although cyclooxygenase (COX)-2 expression was not affected. CHE also suppressed LPS-induced IκB kinase (IKK), IκB, and p65 phosphorylation in RAW264.7 cells. Western blot and immunofluorescence assays of cytosol and nuclear p65 and the catalytic subunit of NF-κB showed that CHE suppressed LPS-induced p65 translocation from the cytosol to the nucleus. CHE also suppressed LPS-induced Interleukin (IL)-6 and tumor necrosis factor (TNF)-α production in RAW264.7 cells. These results suggest that CHE prevents NO-mediated inflammation by suppressing NF-κB and inflammatory cytokines.

In vitro combinatorial anti-proliferative and immunosuppressive effects of <i>Brucea javanica</i> extract with CX-4945 and imatinib in human T-cell acute lymphoblastic leukemia cells

Jung, Jung-Il,Kim, Se Young,Park, Kyeong-Yong,Sydara, Kongmany,Lee, Sang Woo,Kim, Soon Ae,Kim, Jiyeon Elsevier 2018 BIOMEDICINE AND PHARMACOTHERAPY Vol.106 No.-

<P><B>Abstract</B></P> <P>Since 1970, the isolated and identified components of <I>Brucea javanica</I> (L.) Merr. have been known to contain anticancer effects, particularly antileukemic effect. In this study, the inhibitory effect of <I>Brucea javanica</I> (BJ) on cell growth and inflammation was confirmed in human T-cell acute lymphocytic leukemia (T-ALL) cells, and its efficacy as an antileukemic agent was verified. Our results showed that BJ extract induced caspase-dependent apoptosis of T-ALL Jurkat cells through inhibition of the CK2-mediated signaling pathway, while exerting no significant cytotoxicity in normal peripheral blood mononuclear cells. Moreover, BJ extract suppressed the NF-κB signaling pathway, thus, inhibiting the interleukin (IL)-2 expression induced by phorbol 12-myristate 13-acetate (PMA) and phytohemagglutinin (PHA). Notably, combined treatment with BJ extract plus CX-4945 or imatinib exerted enhanced inhibitory effects on T-ALL cell growth and IL-2 production. Overall, these results suggest that BJ extract can be a potent therapeutic herbal agent for T-ALL treatment and prevention of IL-2 mediated inflammatory immune responses.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>Brucea javanica</I> extract induces caspase-dependent apoptosis of T-ALL cells. </LI> <LI> <I>Brucea javanica</I> extract the PMA/PHA-mediated NF-κB signaling pathway and IL-2 production. </LI> <LI> <I>Brucea javanica</I> extract suppresses expression of CK2 subunits and Akt phosphorylation. </LI> <LI> Combined treatment exerts enhanced inhibitory effects on T-ALL cell viability and IL-2 production. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Streptococcus mutans에 대한 인도감나무 줄기 추출물의 항균활성 및 생물막 형성 억제 효과

김혜수(Hye Soo Kim),이상우(Sang Woo Lee),콩마니 시다래(Kongmany Sydara),조수정(Soo Jeong Cho) 한국생명과학회 2019 생명과학회지 Vol.29 No.1

본 연구에서는 천연물 유래 구강 건강소재로써 인도감나무 줄기 추출물의 이용 가능성을 평가하기 위해 TLC, TLC-bioautography, HPLC, ESI-MS 등을 이용하여 인도감나무 줄기 추출물로부터 S. mutans KCTC3065에 대해 항균활성이 있는 항균물질을 분리하고 주사전자현미경과 real-time PCR을 이용하여 추출물이 S. mutans의 생물막에 미치는 영향을 조사하였다. S. mutans에 대한 인도감나무 줄기 추출물의 항균활성은 극성이 낮은 n-hexane 분획물에서 확인되었고 TLC, TLC-bioautography, HPLC에 의해 분리된 항균물질의 분자량은 ESI-MS분석 결과 188로 추정되었다. 인도감나무 줄기 추출물(1 mg/ml) 처리에 따른 S. mutans의 바이오필름 바이오매스 변화는 주사전자현미경으로 관찰하였으며 추출물을 처리하지 않은 대조구는 추출물 처리구에 비해 세포가 군집을 이루고 모여 있었으며 세포 주변에서 바이오필름이 관찰되었지만 추출물을 처리한 처리구의 세포 주변에서는 바이오필름을 관찰할 수 없었다. Real-time PCR을 이용하여 바이오필름 생성 과정에서 치면 부착에 필수적인 GTFs의 발현양상을 조사한 결과, 인도감나무 추출물이 0.2-1.0 mg/ml의 농도로 처리된 배양액에서 gtfB 유전자 발현은 추출물의 농도에 따라 큰 변화가 없었지만 gtfC 유전자 발현은 추출물의 농도가 높아질수록 감소하는 경향을 나타내었다. 이상의 결과를 종합하면 인도감나무 줄기 추출물은 바이오필름 형성 단계에서 치아우식증 원인균인 S. mutans의 초기 치면 부착을 저해함으로서 바이오필름 생성을 억제할 수 있는 천연물 유래 구강 건강소재로써 이용 가능성이 높을 것으로 판단된다. The objective of this study was to evaluate the potential of Diospyros malabarica stem extract, a natural materials, in oral health material. With this aim in mind, thin layer chromatography (TLC), TLC-bioautography, high-performance liquid chromatography (HPLC), electrospray ionization–mass spectrometry (ESI-MS), scanning electron microscopy (SEM), and real-time qPCR were performed. The antibacterial activity of D. malabarica stem extract against Streptococcus mutans KCTC3065 was confirmed in an n-hexane fraction with low polarity. The molecular weight of the antibacterial compound was estimated to be 188 by ESI-MS analysis. The inhibitory effects of the extract on biofilm formation and gene expression related to biofilm formation of S. mutans were determined by SEM and real-time PCR analysis. The extract inhibited the formation of S. mutans biofilms at D. malabarica stem extract concentrations of 1 mg/ml, as shown by SEM. The real-time PCR analysis showed that the expression of the gtfC gene, which is associated with biofilm formation, was significantly decreased in a dose-dependent manner. Based on the above results, it can be concluded that D. malabarica stem extracts, a natural materials, can be used in oral health products to suppress the formation of biofilms by inhibiting tooth adhesion of S. mutans, a causative agent of dental caries.

인도감나무 줄기 추출물이 구강미생물의 생육과 바이오필름 생성에 미치는 영향

김혜수(Hye Soo Kim),권현숙(Hyun Sook Kwon),김철환(Chul Hwan Kim),이상우(Sang Woo Lee),콩마니 시다래(Kongmany Sydara),조수정(Soo Jeong Cho) 한국생명과학회 2018 생명과학회지 Vol.28 No.1

본 연구는 천연물유래 구강건강소재로써 약용작물의 이용 가능성을 알아보기 위해 수행되었으며 네팔, 라오스, 몽골, 방글라데시, 베트남, 중국 등지에 자생하는 200여 종의 약용작물로부터 추출한 메탄올 추출물 중 인도감나무 줄기 추출물(1 mg/disc)이 P. gingivalis ATCC33277와 S. mutans ATCC25175에 대해 가장 우수한 항균활성을 나타내었으며 양성대조구로 사용한 chlorhexidine, sodium lauryl sulfate, triclosan과 유사한 항균활성을 나타내었다. 인도감나무 줄기 추출물은 P. gingivalis ATCC33277에 비해 S. mutans ATCC25175에 대해 높은 항균활성을 나타내었으며 P. gingivalis ATCC33277에 대해서는 살균작용(MBC, 0.4 mg/ml)을, S. mutans ATCC25175에 대해서는 정균작용을 하는 것으로 확인되었다. 인도감나무 줄기 추출물이 0.2-1.0 mg/ml 농도로 처리된 배양액에서 S. mutans ATCC25175의 바이오필름 생성율과 바이오필름 생성관련 유전자 comX의 발현은 추출물의 농도가 높아질수록 억제되는 것을 확인할 수 있었다. 이상의 결과를 종합하면 인도감나무 줄기 추출물은 치아우식증 원인균인 S. mutans ATCC25175에 대한 정균작용과 바이오필름 생성 억제능이 우수하기 때문에 천연물유래 구강건강소재로써 이용 가능성이 높을 것으로 판단된다. This study was conducted to investigate the potential of medicinal plants as oral health materials derived from natural products. Among the extracts from 200 medicinal plants grown in Nepal, Laos, Mongolia, Bangladesh, Vietnam, and China, stem extracts from Diospyros malabarica (1 mg/disc) showed the highest antibacterial activity against Porphyromonas gingivalis ATCC33277 and Streptococcus mutans ATCC25175. The D. malabarica stem extracts showed antibacterial activity similar to chlorhexidine, sodium lauryl sulfate, and triclosan, which were used as a positive control, as well as higher antibacterial activity against S. mutans ATCC25175 than P. gingivalis ATCC33277. The D. malabarica stem extracts showed bactericidal action (MBC, 0.4 mg/ml) against P. gingivalis ATCC33277 and bacteriostatic action against S. mutans ATCC25175. The biofilm production rate of S. mutans ATCC25175 and the expression of the comX gene associated to biofilm formation in the cultures treated with 0.2-1.0 mg/ml of D. malabarica stem extracts were suppressed in a concentration-dependent manner. Based on the above results, it can be concluded that D. malabarica stem extracts can be used as oral health material derived from natural materials, as demonstrated by the bacteriostatic action and inhibition of biofilm formation against S. mutans ATCC25175.

Propionibacterium acnes에 의해 유도되는 염증반응에서 Cnestis palala (Lour.) Merr. 추출물의 억제효과

신진학,이은혜,김선숙,서수련,Shin, Jin Hak,Lee, Eun Hye,Kim, Seon Sook,Sydara, Kongmany,Seo, Su Ryeon 한국미생물학회 2018 미생물학회지 Vol.54 No.1

여드름은 사춘기와 젊은 연령층에서 나타나는 염증성 피부질환이다. Propionibacterium acnes (P. acnes)는 여드름에서 염증을 유발하는 주요한 균으로 알려져 있다. P. acnes 는 피부에서 과잉생산된 피지에 의해 막힌 모낭 내에서 증식하며, 피부 내의 단핵구를 활성화시키고 염증성 사이토카인(pro-inflammatory cytokine)의 분비를 촉진하여 염증반응을 증가시키는 것으로 알려져 있다. 본 논문에서는 Cnestis palala (Lour.) Merr. 추출물이 P. acnes 에 의한 염증반응을 감소시킬 수 있는지 확인하고자 하였다. 마우스 대식세포주인 RAW264.7 에서 C. palala 추출물은 P. acnes 에 의해 증가하는 염증성 사이토카인인 $IL-1{\beta}$의 단백질 발현량과 IL-6, $TNF-{\alpha}$, COX-2 의 mRNA 발현량을 감소시켰다. 또한 염증성 사이토카인 발현의 주요 전사인자(transcription factor)인 $NF-{\kappa}B$ 의 전사 활성 역시 C. palala 추출물에 의해 감소하는 것을 확인하였다. 따라서 C. palala 추출물이 P. acnes 에 의해 발생하는 여드름의 치료제나 그 보조제로 사용될 가능성이 있음을 제안 할 수 있다. Acne is an inflammatory skin disease that occurs in puberty and young people. Propionibacterium acnes (P. acnes) is known to be a major cause of inflammation in acne. P. acnes proliferates within hair follicles blocked by overproduced sebum in the skin, and thereby activates monocytic cells to promote the secretion of pro-inflammatory cytokines. In this study, we investigated the possibility of Cnestis palala (Lour.) Merr. extract to diminish P. acnes-mediated inflammatory responses. We found that C. palala extract significantly attenuated P. acnes-induced pro-inflammatory cytokine expressions, such as $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, iNOS, and COX-2 in mouse macrophage RAW264.7 cells. Moreover, we observed that C. palala extract inhibited $NF-{\kappa}B$ transcriptional activation, which is the major transcription factor of inflammatory cytokine expression. Therefore, it is expected that C. palala extract has a potential as a therapeutic agent or supplement for the treatment P. acnes-induced inflammatory responses.