Effects of Synthetic Lysophosphatidylcholines on Suspension Cultures of the Chinese Hamster Ovary DG44 Cells in Protein-free Media

김인경,임동진,박홍우 한국생물공학회 2008 Biotechnology and Bioprocess Engineering Vol.13 No.4

This study described the effects of synthetic lysophosphatidylcholines on the growth of recombinant CHO-DG44 cells in suspension. Overall, cell growth characteristics were improved when cultivated in suspension in a protein-free medium supplemented with natural soybean lysophosphatidylcholines. To substitute synthetic lysophosphatidylcholines for the naturally occurring lysophosphatidylcholines, we implemented a systematic approach in which twelve synthetic lysophosphatidylcholines were grouped into three lipid mixtures according to the length of their acyl chains. We found that synthetic lysophosphatidylcholines with medium acyl chain lengths (C14-C18), including oleoyl lysophosphatidylcholine (C18:1) could increase cell growth in the protein-free media. The fortified protein-free medium with medium acyl chain length lysophosphatidylcholines (C14-C18) maintained growth of CHO-DG44 cells over five consecutive passages, whereas the cell growth in a CHO protein-free medium was decreased gradually after four passages. We also observed that the restorative effect of oleoyl lysophosphatidylcholine was comparable to that of natural lysophosphatidylcholine in batch and long-term cultivation. These results show that synthetic lysophosphatidylcholines can be used as lipid supplements in either protein-free media or chemi-cally defined media for CHO cell suspension cultures.

Effects of chlorogenic acid on intracellular calcium regulation in lysophosphatidylcholine-treated endothelial cells

( Hye-jin Jung ),( Seung-soon Im ),( Dae-kyu Song ),( Jae-hoon Bae ) 생화학분자생물학회(구 한국생화학분자생물학회) 2017 BMB Reports Vol.50 No.6

Lysophosphatidylcholine (LPC) is a major phospholipid com-ponent of oxidized low-density lipoprotein (ox-LDL) and is implicated in its atherogenic activity. This study investigated the effects of LPC on cell viability, intracellular calcium homeostasis, and the protective mechanisms of chlorogenic acid (CGA) in human umbilical vein endothelial cells (HUVECs). LPC increased intracellular calcium ([Ca²⁺]i) by releasing Ca²⁺ from intracellular stores and via Ca²⁺ influx through store-operated channels (SOCs). LPC also increased the generation of reactive oxygen species (ROS) and decreased cell viability. The mRNA expression of Transient receptor potential canonical (TRPC) channel 1 was increased signifi-cantly by LPC treatment and suppressed by CGA. CGA inhibited LPC-induced Ca²⁺ influx and ROS generation, and restored cell viability. These results suggested that CGA inhibits SOC-mediated Ca²⁺ influx and ROS generation by attenuating TRPC1 expression in LPC-treated HUVECs. Therefore, CGA might protect endothelial cells against LPC injury, thereby inhibiting atherosclerosis. [BMB Reports 2017; 50(6): 323-328]

토끼 관동맥 평활근 세포의 K⁺ 전류 활성에 미치는 Lysophosphatidylcholine의 효과

이창헌,하미영,안덕선,강복순 대한순환기학회 2001 Korean Circulation Journal Vol.31 No.2

Lysophosphatidylcholine의 혈관평활근세포에 대한 세포 독성

강영희 ( Young Hee Kang ),이영주 ( Young Joo Lee ),이동윤 ( Dong Yun Lee ),유미라 ( Mee Ra Rhyu ),최두석 ( Doo Seok Choi ),윤병구 ( Byung Koo Yoon ) 대한폐경학회 2012 대한폐경학회지 Vol.18 No.3

Objectives: To investigate the cytotoxic effects of lysophosphatidylcholine (lysoPC), an active component of oxidized low-density lipoproteins (LDL), on vascular smooth muscle cells (VSMCs). Methods: VSMCs were derived from rat aorta. Cell death was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, lactic dehydrogenase (LDH) assay, and DNA fragmentation assay. Apoptosis was quantified by propidium iodide staining and fluorescent activated cell sorting (FACS) analysis, and intracellular free radical production was determined using 2``, 7``-dichlorofluorescin diacetate (DCF-DA). In addition, the changes in caspases, bcl-2 and bax proteins were evaluated by western blot analysis. Results: LysoPC over 25 μM induced more than 50% of the cell death at 10 hours on MTT assay with no change in the level of LDH. The DNA ladder pattern showed that cell death induced by lysoPC was caused by apoptosis, which was associated with increased free radical production. Vitamin E, a potent antioxidant and caffeic acid phenylethyl ester (CAPE), an inhibitor of nuclear factor-kappaB (NF-kB), blocked apoptosis. The casepase-3 precursor decreased and the active form of caspase-8 increased. Total bcl-2 and bax proteins did not change with lysoPC treatment, but translocation of bax from cytosole to the mitochondria membrane was observed. Conclusion: LysoPC induces apoptosis in VSMCs via an oxidant mechanism, dependent on NF-kB. (J Korean Soc Menopause 2012;18:139-146)

춘계학술대회 : 구연 ; 담관세포에서 Raf-1 활성을 통한 Lysophosphatidylcholine의 cyclooxygenase-2 발현 유도 및 항자멸사 효과

윤정환,곽금연,김윤준,이효석,김정룡 대한간학회 2005 Clinical and Molecular Hepatology(대한간학회지) Vol.11 No.3(S)

배경/목적: 최근 담관세포암의 진단 및 치료에 관한 많은 연구와 발전이 이루어져 왔지만 아직 그 예후는 불량하며 수술 이외의 효과적인 근치적 치료법은 없는 상황으로, 효과적인 치료약제 개발을 위한 담관세포암의 발생 및 진행과 관련된 원인적 기전 규명이 필수적이다. 한편, 담관세포암의 선행 질환으로는 췌담도 합류 이상이 잘 알려져 있으며 췌담도 합류 이상의 구조적 이상에 동반되는 담즙의 구성 성분 변화가 그 원인적 인자로 거론되고 있으나 구체적인 기전은

초록집 : 구연 ; 담관세포에서 Raf-1 활성을 통한 Lysophosphatidylcholine의 cyclooxygenase-2 발현 유도 및 항자멸사 효과

윤정환,곽금연,김윤준,이효석,김정룡 대한간학회 2005 춘계 학술대회 Vol.11 No.-

Betaine attenuates lysophosphatidylcholine-mediated adhesion molecules in aged rat aorta: Modulation of the nuclear factor-κB pathway

Lee, E.K.,Jang, E.J.,Jung, K.J.,Kim, D.H.,Yu, B.P.,Chung, H.Y. Pergamon Press ; Elsevier Science Ltd 2013 Experimental Gerontology Vol.48 No.5

We previously reported that lysophosphatidylcholine (LPC) is a mediator of endothelial dysfunction in the expression of adhesion molecules (AMs) during aging. This study aimed at investigating the effects of betaine on LPC-related expression of AMs and the molecular modulation of nuclear factor-κB (NF-κB) activation in the aorta of aged rats and rat endothelial YPEN-1 cells. The experiment was performed on young (7months) and old (21months) rats; 2 groups of old rats were fed betaine (3 or 6mg.kg<SUP>-1</SUP>.day<SUP>-1</SUP> for 10days). Betaine inhibited the expression of LPC-related AMs in the serum and tissue of aged rats, without affecting the elevated levels of serum LPC. Betaine also prevented the generation of reactive species, thereby maintaining the redox status via the enhancement of the thiol status during aging. Furthermore, betaine attenuated NF-κB activation via the dephosphorylation of IκB kinase (IKK) and mitogen-activated protein kinases (MAPKs) in aged aorta and LPC-treated YPEN-1 cells. Thus, betaine suppressed the LPC-related AM expression associated with NF-κB activation via the upregulation of IKK/MAPKs. Our findings provide insights into the prevention of vascular disorders and the development of interventions based on natural compounds, such as betaine.

Lysophosphatidylcholine aggravates contact hypersensitivity by promoting neutrophil infiltration and IL17 expression

( Mi Hye Song ),( Anupriya Gupta ),( Hye One Kim ),( Kwonik Oh ) 생화학분자생물학회(구 한국생화학분자생물학회) 2021 BMB Reports Vol.54 No.4

Lysophosphatidylcholine (LPC) is a bioactive lysolipid known to contribute to the development of lung allergic diseases. However, it remains unknown whether LPC possesses proinflammatory properties in the skin as well. Here, we investigated this issue by injection of LPC into the murine contact hypersensitivity (CHS) model induced by 2,4-dinitrofluorobenzene (DNFB). LPC increased the expression of IL17, recruited more neutrophils, and eventually aggravated the CHS in the skins. Moreover, the effects of LPC diminished after neutralizing IL17 or depleting neutrophils. Mechanistically, LPC upregulated not only IL17 but also CXCL1 and CXCL2 in a G2A-dependent manner. Taken together, our study demonstrated that the upregulation of LPC could contribute to allergic skin inflammation by increasing IL17 expression and neutrophil recruitment via G2A receptor. [BMB Reports 2021; 54(4): 203-208]

Lysophosphatidylcholine Increases Ca²+ Current via Activation of Protein Kinase C in Rabbit Portal Vein Smooth Muscle Cells

정승수,이영호,한성식,김영환,남택상,안덕선 대한약리학회 2008 The Korean Journal of Physiology & Pharmacology Vol.12 No.1

Lysophosphatidylcholine (LPC), a metabolite of membrane phospholipids by phospholipase A2, has been considered responsible for the development of abnormal vascular reactivity during atherosclerosis. Ca²+ influx was shown to be augmented in atherosclerotic artery which might be responsible for abnormal vascular reactivity. However, the mechanism underlying Ca²+ influx change in atherosclerotic artery remains undetermined. The purpose of the present study was to examine the effects of LPC on L-type Ca²+ current (ICa(L)) activity and to elucidate the mechanism of LPC-induced change of ICa(L) in rabbit portal vein smooth muscle cells using whole cell patch clamp. Extracellular application of LPC increased ICa(L) through whole test potentials, and this effect was readily reversed by washout. Steady state voltage dependency of activation or inactivation properties of ICa(L) was not significantly changed by LPC. Staurosporine (100 nM) or chelerythrine (3μM), which is a potent inhibitor of PKC, significantly decreased basal ICa(L), and LPC-induced increase of ICa(L) was significantly suppressed in the presence of PKC inhibitors. On the other hand, application of PMA, an activator of PKC, increased basal ICa(L) significantly, and LPC-induced enhancement of ICa(L) was abolished by pretreatment of the cells with PMA. These findings suggest that LPC increased ICa(L) in vascular smooth muscle cells by a pathway that involves PKC, and that LPC-induced increase of ICa(L) might be, at least in part, responsible for increased Ca²+ influx in atherosclerotic artery. Lysophosphatidylcholine (LPC), a metabolite of membrane phospholipids by phospholipase A2, has been considered responsible for the development of abnormal vascular reactivity during atherosclerosis. Ca²+ influx was shown to be augmented in atherosclerotic artery which might be responsible for abnormal vascular reactivity. However, the mechanism underlying Ca²+ influx change in atherosclerotic artery remains undetermined. The purpose of the present study was to examine the effects of LPC on L-type Ca²+ current (ICa(L)) activity and to elucidate the mechanism of LPC-induced change of ICa(L) in rabbit portal vein smooth muscle cells using whole cell patch clamp. Extracellular application of LPC increased ICa(L) through whole test potentials, and this effect was readily reversed by washout. Steady state voltage dependency of activation or inactivation properties of ICa(L) was not significantly changed by LPC. Staurosporine (100 nM) or chelerythrine (3μM), which is a potent inhibitor of PKC, significantly decreased basal ICa(L), and LPC-induced increase of ICa(L) was significantly suppressed in the presence of PKC inhibitors. On the other hand, application of PMA, an activator of PKC, increased basal ICa(L) significantly, and LPC-induced enhancement of ICa(L) was abolished by pretreatment of the cells with PMA. These findings suggest that LPC increased ICa(L) in vascular smooth muscle cells by a pathway that involves PKC, and that LPC-induced increase of ICa(L) might be, at least in part, responsible for increased Ca²+ influx in atherosclerotic artery.

Novel Approach for Analysis of Bronchoalveolar Lavage Fluid (BALF) Using HPLC-QTOF-MS-Based Lipidomics: Lipid Levels in Asthmatics and Corticosteroid-Treated Asthmatic Patients

Kang, Yun Pyo,Lee, Won Jun,Hong, Ji Yeon,Lee, Sae Bom,Park, Jeong Hill,Kim, Donghak,Park, Sunghyouk,Park, Choon-Sik,Park, Sung-Woo,Kwon, Sung Won American Chemical Society 2014 JOURNAL OF PROTEOME RESEARCH Vol.13 No.9

<P>To better understand the respiratory lipid phenotypes of asthma, we developed a novel method for lipid profiling of bronchoalveolar lavage fluid (BALF) using HPLC-QTOF-MS with an internal spectral library and high-throughput lipid-identifying software. The method was applied to BALF from 38 asthmatic patients (18 patients with nonsteroid treated bronchial asthma [NSBA] and 20 patients with steroid treated bronchial asthma [SBA]) and 13 healthy subjects (NC). We identified 69 lipids, which were categorized into one of six lipid classes: lysophosphatidylcholine (LPC), phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylserine (PS), sphingomyelin (SM) and triglyceride (TG). Compared with the NC group, the individual quantity levels of the six classes of lipids were significantly higher in the NSBA subjects. In the SBA subjects, the PC, PG, PS, SM, and TG levels were similar to the levels observed in the NC group. Using differentially expressed lipid species (<I>p</I> value < 0.05, FDR < 0.1 and VIP score of PLS-DA > 1), 34 lipid biomarker candidates with high prediction performance between asthmatics and controls were identified (AUROC > 0.9). These novel findings revealed specific characteristics of lipid phenotypes in asthmatic patients and suggested the importance of future research on the relationship between lipid levels and asthma.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jprobs/2014/jprobs.2014.13.issue-9/pr5002059/production/images/medium/pr-2014-002059_0008.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/pr5002059'>ACS Electronic Supporting Info</A></P>