[논문]과루인 및 과루인청혈플러스가 인간 탯줄 정맥 내피 세포(Human umbilicl vein endothelial cells, HUVECs)에서 염증인자 발현 억제에 미치는 영향

김해융; 설인찬; 유호룡; 김윤식

doi:10.22246/jikm.2022.43.4.514

과루인 및 과루인청혈플러스가 인간 탯줄 정맥 내피 세포(Human umbilicl vein endothelial cells, HUVECs)에서 염증인자 발현 억제에 미치는 영향
The Effect of Trichosanthes Kirilowii Maximowicz Extract and Trichosanthes Kirilowii Maximowicz Cheonghyeol Plus on Anti-Inflammatory Factor Expression in Human Umbilical Vein Endothelial Cells (HUVECs) 원문보기

대한한방내과학회지 = The journal of internal Korean medicine, v.43 no.4, 2022년, pp.514 - 528

김해융 (대전대학교 한의과대학 심계내과학교실) , 설인찬 (대전대학교 한의과대학 심계내과학교실) , 유호룡 (대전대학교 한의과대학 심계내과학교실) , 김윤식 (대전대학교 한의과대학 심계내과학교실)

Abstract ▼ AI-Helper

Objective: To examine the effects of Trichosanthes kirilowii Maximowicz extract (TE) and Trichosanthes kirilowii Maxi mowicz Cheonghyeol Plus Phellinus linteus Cheonghyeol plus (TCP) on anti-inflammatory factor expression in human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were activated with TNF-α and then treated with TE and TCP. The expression levels were then measured for intracellular genes (KLF2, eNOS, MCP-1, ICAM-1, and VCAM-1), proteins (KLF2, eNOS, MCP-1, ICAM-1, VCAM-1, ERK, and JNK, p38), and extracellular biomarkers (ICAM-1, VCAM-1, and MCP-1). Results: 1. TCP at concentrations of 100 ㎍/mL or greater significantly increased the expression of KLF2 and eNOS intracellular genes and significantly decreased the expression of ICAM-1, VCAM-1, and MCP-1 genes compared to the control group. 2. TCP at concentrations of 100 ㎍/mL or greater significantly increased the expression of KLF2, eNOS proteins compared to the control group, and significantly reduced the expression of VCAM-1, ICAM-1, MCP-1, ERK, and p38 proteins. However, JNK protein phosphorylation showed no significant change compared to the control group. 3. TCP at concentrations of 100 ㎍/mL or more significantly decreased the production of MCP-1, ICAM-1, and VCAM-1 extracellular biomarkers compared to the control group. 4. TE at a concentration of 100 ㎍/mL did not cause any significant change in the expression of intracellular genes or proteins, in the production of the extracellular biomarker MCP-1, or in the amount of JNK protein compared to the control group. Other intracellular genes, proteins, and extracellular biomarker expression showed the same trend as observed with TCP exposure. Conclusion: This study experimentally confirmed that TE and TCP could be effective in preventing or inhibiting various inflammatory vascular diseases due to their anti-inflammatory effects.

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표/그림 (9)

표 Table 1. The Prescription of TE or TCP
표 Table 2. Real-Time PCR Primer Sequences
그림 Fig. 1. Cell viability of TE or TCP in HUVEC. HUVEC were treated by TE (100 ug/mL) or TCP (100, 200, 400, and 600 ug/mL) for 24 hours. Treated cells were reacted by EZ-Cytox for 30 minutes and absorbance was measured at 450 nm using micro plate reader. Cell viability was calculated as percentage relative to the control group. The results were presented by the mean±standard error of mean from three independent experiments (Significance of results, ** ; p<0.01 compared to the control).
그림 Fig. 2. Effect of TE or TCP on KLF2 (A), eNOS (B) mRNA expression level in HUVECs. HUVECs were treated TE (100 ug/mL) or TCP (100, 200, and 400 ug/mL), with TNF-α 10 ng/mL for 12 h. The mRNA expression level was measured using a real-time PCR (qPCR). The results were presented by the mean±standard error of mean from three independent experiments (Significance of results, +++ ; p<0.001 compared to the normal group, ** ; p<0.01, *** ; p<0.001 compared to the control group).
그림 Fig. 3. Effect of TE or TCP on MCP-1 (A), ICAM-1 (B), VCAM-1 (C) mRNA expression level in HUVECs. HUVECs were treated TE (100 ug/mL) or TCP (100, 200, and 400 ug/mL), with TNF-α 10 ng/mL for 12 h. The mRNA expression level was measured using a real-time PCR (qPCR). The results were presented by the mean±standard error of mean from three independent experiments (Significance of results, +++ ; p<0.001 compared to the normal group, ** ; p<0.01, *** ; p<0.001 compared to the control group).
그림 Fig. 4. Effect of TE or TCP on KLF2 (A), eNOS (B) protein expression level in HUVECs. HUVECs were treated TE (100 ug/mL) or TCP (100, 200, and 400 ug/mL), with TNF-α 10 ng/mL for 12 hours. The protein expression level was measured using a western blot. The results were presented by the mean±standard error of mean from three independent experiments (Significance of results, +++ ; p<0.001 compared to the normal, *** ; p<0.001 compared to the control).
그림 Fig. 5. Effect of TE or TCP on MCP-1 (A), ICAM-1 (B), VCAM-1 (C), ERK (D), p38 (E) protein expression level in HUVECs. HUVECs were treated TE (100 ug/mL) or TCP (100, 200, and 400 ug/mL), with TNF-α 10 ng/mL for 12 hours. The protein expression level was measured using a western blot. The results were presented by the mean±standard error of mean from three independent experiments (Significance of results, +++ ; p<0.001 compared to the normal, *** ; p<0.001 compared to the control).
그림 Fig. 6. Effect of TE or TCP on JNK protein expression level in HUVECs. HUVECs were treated TE (100 ug/mL) or TCP (100, 200, and 400 ug/mL), with TNF-α 10 ng/mL for 12 hours. The protein expression level was measured using a western blot. The results were presented by the mean±standard error of mean from three independent experiments (Significance of results, +++ ; p<0.001 compared to the normal, *** ; p<0.001 compared to the control).
그림 Fig. 7. Effect of TE or TCP on MCP-1 (A), ICAM-1 (B), VCAM-1 (C) level in HUVECs. HUVECs were treated TE (100 ug/mL) or TCP (100, 200, and 400 ug/mL), with TNF-α 10 ng/mL for 12 hours. MCP-1 level was measured using a ELISA kit. The results were presented by the mean±standard error of mean from three independent experiments (Significance of results, +++ ; p<0.001 compared to the normal, * ; p<0.05, ** ; p<0.01, *** ; p<0.001 compared to the control).

참고문헌 (29)

Jung IK. Review Articles : Molecular Biology of Atherosclerosis. Endocrinology and Metabolism 2010;25(3):166-70.
Glass CK, Witztum JL. Atherosclerosis : The road ahead. Cell 2001;104(4):503-16.

상세보기
Kim CJ. Atherosclerosis and Inflammation. Korean Journal of Lipidology 2001;11(4):413-9.
Yim CY. Review : Nitric oxide and cancer. Korean Journal of Medicine 2010;78(4):430-6.
Lee HR, Kim SW, Yoo M. Analysis of Single Nucleotide Polymorphism of eNOS Genes in Korean Genome. Journal of Life Science 2014;24(2):181-5.
Kim MJ, Kim KBWR, Park SH, Choi JS. Anti-Inflammatory Effect of Chondria crassicaulis Ethanol Extract onMAPKs and NF-κB Signaling Pathway in LPS-Induced RAW 264.7 Macrophages. KSBB Journal 2017;32(4):352-60.
Won SY, Yoo HR, Seol IC, Kim YS. The Effect of Phellinus Linteus Cheonghyeol Plus on Inhibition of Antioxidant and Inflammatory Factor Expression Associated with Dyslipidemia in Human Umbilical Vein Endothelial Cells (HUVECs). J Korean Med 2020;41(2):43-57.
Chae IC, Ryu JY, Yoo HR, Kim YS, Seol IC. The Anti-dyslipidemia Effects of Allii Macrostemonis Bulbus Cheonghyeol Plus in HUVEC by Antioxidant and Anti-inflammatory activity. J Physiol &Pathol Korean Med 2020;34(3):126-35.
Lee JG, Kim YH, Lee DN, Kim HJ. Growth Arrest and Apoptosis of Human Uterine Cervical Carcinoma Cells Induced by Trichosanthes Semen Extract. Journal of Physiology & Pathology in Korean Medicine 2005;19(4):965-72.
신민교. 임상본초학. 제1판. 서울: 영림사; 1997, p. 369-72.
Son JH, Kim DC. In Vitro Anti-Bacterial and Anti-Inflammatory Effects of Trichosanthes Semen, Gardeniae Fructus, and Angelicae Dahuricae Radix Aqueous Extracts. The journal of oriental obstetrics & gynecology 2013;26(1):41-58.
Statistics Korea. Causes of death statistics in Korea. Daejeon: Statistics Korea; 2020.
Ito T, Ikeda U. Inflammatory cytokines and cardiovascular disease. Curr Drug Targets Inflamm Allergy 2003;2(3):257-65.
Cybulsky MI, Liyama K, Li H, Zhu S, Chen M, Liyama M, et al. A major role for VCAM-1, but not ICAM-1, in early atherosclerosis. J Clin Invest 2001;107(10):1255-62.

상세보기
한기훈. 대식세포와 동맥경화. 한국지질동맥경 화학회 2009;2009(0):80-2.
Youn JS, Kim MS, Na HJ, Jung HR, Song CK, Kang SY, et al. Screening test for Dendropanax morbifera Leveille extracts : in vitrocomparison to ox-LDL-induced lipid accumulation, ethanolinduced fatty liver and HMG-CoA reductase inhibition. Journal of Applied Biological Chemistry Korean Society for Applied Biological Chemistry 2018;61(1):1-8.
Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins: identificationby morphologic and immunologic criteria. J Clin Invest 1973;52(11):2745-56.

상세보기
Turpaev KT. Transcription Factor KLF2 and Its role in the regulation of inflammatory process. Biochemistry Moscow 2020;85(1):54-67.
Song EY, Cho IH, Cho JY. The Effects of Exercise Training on Cardiac ENOS, ET-1 MRNA and Skeletal Muscle ENOS Protein Level in SHR. Journal of Life Science 2007;17(12):1717-22.
Lim HC, Jeon SY. The effect of Gamidohongsamultang on the Gene Expression Levels of eNOS, KLF2, ICAM-1 and VCAM-1 in HUVEC Cell. J Int Korean Med 2018;39(1):1-8.
Power CA. Proudfoot AE. The chemokine system : novel broad spectrum therapeutic targets. Curr Opin Pharmacol 2001;1(4):417-24.

상세보기
Takeya M, Yoshimura T, Leonard EJ, Takahashi K. Detection of monocyte chemoattratant protein-1 in human atherosclerotic lesions by anti-monocyte chemoattractant protein-1 in monoclonal antibody. Hum Pathol 1993;24(5):534-9.
Seino Y, Ikeda U, Sekiguchi H, Takahashi M, Hojo Y, Irokawa M, et al. Expression of monocyte chemoattactant protein-1 in vascular tissue. Cytokine 1995;7(6):575-9.
Kim SH, Kim SH, Ryu SR, Lee PJ, Moon C. Inhibitory Effects of Zerumbone on MCP-1 induced THP-1 Migration. Korean Journal of Clinical Lab Sci 2018;50(2):177-82.
Park HS, Kim KS, Park CW, Kim MJ, Cho YK, Lee SW, et al. The relationship between Progression of Coronary Artery Stenosis and Serum adiponectin, ICAM-1(Intercellular Adhesion Molecule-1) Level. Korean Circulation Journal 2004;34(9):837-44.
Elice MJ, Osborn L, Takata Y, Crouse C, Stefan L, Martin E, et al. VACAM-1 on activated endothelium interacts with the leukocyte integrin VAL-4 at a site distinct from the VLA-4/fibronectin binding site. Cell 1990;60(4):577-84.
Su B, Karin M. Mitogen-activated protein kinase cascades and regulation of gene expression. Curr Opin Immunol 1996;8(3):402-11.

상세보기
Chen YR, Wang X, Templeton D, Davis RJ, Tan TH. The role of c-Jun N-terminal kinase (JNK) in apoptosis induced by ultraviolet C and gamma radiation. Duration of JNK activation may determine cell death and proliferation. J Biol Cem 1996;271(50):31929-36.
Kawasaki H, Morooka T, Shimohama S, Kimura J, Hirano T, Gotoh Y, et al. Activation and involvement of p38 mitogen-activated protein kinase in glutamate-induced apoptosis in rat cerebellar granule cells. J Biol Chem 1997;272(30):18518-21.

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표제어: PCR

동의어: Packet Collision Rate

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용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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