[논문]금은화가 HUVEC에서 죽상동맥경화증 관련 염증인자 발현 억제에 미치는 영향

양지혜; 유호룡; 김윤식; 설인찬

doi:10.22246/jikm.2021.42.1.25

금은화가 HUVEC에서 죽상동맥경화증 관련 염증인자 발현 억제에 미치는 영향
The Effect of Lonicera Japonica Thunberg on Inflammatory Factor Expression Associated with Atherosclerosis 원문보기

대한한방내과학회지 = The journal of internal Korean medicine, v.42 no.1, 2021년, pp.25 - 39

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

Abstract ▼ AI-Helper

Objective: The purpose of this study was to investigate the effect of Lonicera Japonica Thunberg (LJT) on the inflammatory factor expression associated with atherosclerosis in human umbilical vein endothelial cells (HUVECs). Methods: After treatment with LJT in HUVEC which is treated with TNF-α, we measured the expression levels of biomarkers (MCP-1, ICAM-1, VCAM-1, KLF2, and eNOS), mRNA (CCL2, ICAM1, VCAM1, KLF2, and NOS3), and the proteins (MCP-1, ICAM-1, VCAM-1, KLF2, eNOS, ERK, JNK, and p38). Results: 1. Compared to the control, LJT significantly reduced MCP-1 and VCAM-1 levels at concentrations of 100, 200, and 400 ㎍/ml and ICAM-1 expression at 200 and 400 ㎍/ml compared to the control. It increased KLF2 levels at all three concentrations, but not significantly, while eNOS expression was significantly increased at 400 ㎍/ml. 2. LJT was seen to significantly reduce the expression of CCL2, ICAM1, and VCAM1 mRNA at concentrations of 100, 200, and 400 ㎍/ml compared to the control. In contrast, significantly increased KLF2 and NOS3 mRNA levels were observed at 400 ㎍/ml and at 200 and 400 ㎍/ml, respectively. 3. Compared to the control, LJT significantly reduced the protein expression of MCP-1 and VCAM-1 at 200 and 400 ㎍/ml and of ICAM-1 at 400 ㎍/ml. In addition, it increased both KLF2 and eNOS protein levels at 200 and 400 ㎍/ml. Although LJT did not have an effect on ERK expression in comparison with the control, it significantly reduced JNK levels at 200 and 400 ㎍/ml and p38 levels at 400 ㎍/ml. Conclusions: These results suggest that LJT has an effect on the inhibition of inflammatory factor expression associated with atherosclerosis in HUVECs which could contribute to the prevention of cardiovascular and cerebrovascular diseases.

주제어

표/그림 (9)

표 Table 1. The Sequences of Primers
그림 Fig. 1. Cell viability of LJT in HUVEC. HUVEC were treated by 100, 200, 400, and 600 μg/ml of LJT for 24 hours. Treated cells were reacted by an EZ-Cytox for 30 minutes and then absorbance was measured at 450 nm using a micro plate reader. Cell viability was calculated as percentage (%) compared to the control. The results were presented as the mean±standard deviation from 3 separate trials.
그림 Fig. 2. Effect of LJT on MCP-1 (A), ICAM-1 (B)and VCAM-1 (C) levels in HUVEC. Fig. 2. Effect of LJT on MCP-1 (A), ICAM-1 (B) and VCAM-1 (C) levels in HUVEC. HUVEC were treated 100, 200, and 400 μg/ml of LJT with 10ng/ml of TNF-α for 12 hours. MCP-1, ICAM-1, VCAM-1 levels were measured using an ELISA kit. The results were presented as the mean±standard deviation from 3 separate trials (Significance of results, * ; p<0.05, ** ; p<0.01, *** ; p<0.001 compared to control).
그림 Fig. 3. Effect of LJT on KLF2 (A), eNOS (B) levels in HUVEC. HUVEC were treated 100, 200, and 400 μg/ml of LJT with 10ng/ml of TNF-α for 12 hours. KLF2, eNOS levels were measured using an ELISA kit. The results were presented as the mean±standard deviation from 3 separate trials (Significance of results, ** ; p<0.01 compared to control).
그림 Fig. 4. Effect of LJT on CCL2 (A), ICAM1 (B), VCAM1 (C) mRNA expression levels in HUVEC. HUVEC were treated 100, 200, and 400 μg/ml of LJT with 10 ng/ml of TNF-α for 12 hours. The mRNA expression levels were measured with a qRT-PCR (quantitative real-time PCR). The results were presented as the mean±standard deviation from 3 separate trials (Significance of results, * ; p<0.05, ** ; p<0.01, *** ; p<0.001 compared to control).
그림 Fig. 5. Effect of LJT on KLF2 (A), NOS3 (B) mRNA expression levels in HUVEC. HUVEC were treated 100, 200, and 400 μg/ml of LJT with 10 ng/ml of TNF-α for 12 hours. The mRNA expression levels were measured with a qRT-PCR (quantitative real-time PCR). The results were presented as the mean±standard deviation from 3 separate trials (Significance of results, * ; p<0.05, ** ; p<0.01, *** ; p<0.001 compared to control).
그림 Fig. 6. Effect of LJT on MCP-1 (A), ICAM-1 (B), VCAM-1 (C) protein expression levels in HUVEC. HUVEC were treated 100, 200, and 400 μg/ml of LJT with 10 ng/ml of TNF-α for 12 hours. The protein expression levels were measured using a western blot. The results were presented as the mean±standard deviation from 3 separate trials (Significance of results, *** ; p<0.001 compared to control).
그림 Fig. 7. Effect of LJT on KLF2 (A), eNOS (B) protein expression levels in HUVEC. HUVEC were treated 100, 200, and 400 μg/ml of LJT with 10 ng/ml of TNF-α for 12 hours. The protein expression levels were measured using a western blot. The results were presented as the mean±standard deviation from 3 separate trials (Significance of results, ** ; p<0.01, *** ; p<0.001 compared to control).
그림 Fig. 8. Effect of LJT on ERK (A), JNK (B), p38 (C) protein expression levels in HUVEC. HUVEC were treated 100, 200, and 400 μg/ml of LJT with 10 ng/ml of TNF-α for 12 hours. The protein expression levels were measured using a western blot. The results were presented as the mean±standard deviation from 3 separate trials (Significance of results, *** ; p<0.001 compared to control).

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