[논문]LPS로 유도된 염증모델에 대한 청포축어탕의 억제 효과

홍가경; 이수형; 정현태; 김송백

doi:10.15204/jkobgy.2021.34.4.012

[국내논문] LPS로 유도된 염증모델에 대한 청포축어탕의 억제 효과
Inhibitory Effects of Cheongpochukeo-tang on LPS-induced Inflammation Model 원문보기

大韓韓方婦人科學會誌 = The Journal of Korean obstetrics & gynecology, v.34 no.4, 2021년, pp.12 - 29

홍가경 (원광대학교 한의과대학 한방부인과학교실) , 이수형 (원광대학교 한의과대학 한방부인과학교실) , 정현태 (원광대학교 한의과대학 한방부인과학교실) , 김송백 (원광대학교 한의과대학 한방부인과학교실)

Abstract ▼ AI-Helper

Objective: This study was performed to investigate the inhibitory effect of Cheongpochukeo-tang (CCT) on lipopolysaccharide (LPS)-induced inflammation model. Methods: RAW 264.7 cells were pre-treated with CCT and incubated with LPS (500 ng/ml) after 1 hour. Cell viability was measured by MTT assay to figure out cytotoxicity of CCT. The production of nitric oxide and mRNA expression of pro-inflammatory cytokine were measured. And the activation of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) were examined to figure out molecular mechanisms of CCT's anti-inflammatory effects. In addition, mice survival rate and cytokine levels of serum were observed after treated with CCT. And mice liver tissues were observed and their cytokines levels in liver tissue were measured. Results: CCT did not have cytotoxic effect in RAW 264.7 cells. It inhibited LPS-induced nitric oxide (NO) production, but showed an increase in NO by itself at 2 mg/ml concentration. CCT inhibited mRNA expression of IL-1β, IL-6, TNF-α in a dose dependant and the activaton of MAPKs and NF-κB. In addition, CCT reduced mortality in the LPS-induced mouse model and inhibited production of cytokines in mouse serum and liver tissue. Conclusion: The results suggest that CCT could reduce LPS-induced inflammation by inhibiting MAPKs and NF-κB activaton, NO production, and pro-inflammatory cytokines secretion. Thereby, CCT could be effective medicine for the inflammatory disease.

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표/그림 (10)

표 Table 1. Composition of Cheongpochukeo-tang
표 Table 2. The Primer of IL-1β, IL-6, TNF-α
그림 Fig. 1. The cytotoxicity of Cheongpochukeo -tang (CCT) in RAW 264.7 cells. RAW 264.7 cells were incubated with CCT as indicated concentration. After 24 h, cell viability was measured by MTT assay as described in materials and methods. Data were given as means of values±SD from three independent experiments.
그림 Fig. 2. Effects of Cheongpochukeo-tang (CCT) on LPS-induced NO production in RAW 264.7 cells. RAW 264.7 cell was treated with 0.1, 0.5, 1, 2 (mg/ml) of CCT and LPS (500 ng/ml) for 24 h. NO production was measured by using Griess reagent. Data were given as means of values±SD from three independent experiments. * P<0.05 as compared with saline. †P<0.05 as compared with only-LPS-treated group.
그림 Fig. 3. Effects of Cheongpochukeo-tang (CCT) on the mRNA expression of IL-1β, IL-6 and TNF-α in RAW 264.7 cells. The cells were pre-treated with CCT as indicated concentrations for 1 h, and then incubated with or without LPS (500 ng/ml) for 24 h. The level of cytokine was measured by real time RT-PCR. Data were given as means of values±SD from three independent experiments. * P<0.05 as compared with saline. †P<0.05 as compared with only-LPS-treated group.
그림 Fig. 4. Effects of Cheongpochukeo-tang (CCT) on the activation of MAPKs and NF-κB in LPS-stimulated RAW 264.7 cells. The cells were pre-treated with CCT (1 mg/ml) for 1 h, and then incubated with LPS (500 ng/ml) for indicated mins. Detail methods were described in Materials and Methods. The results shown are representative of three independent experiments. * : phosphorylated extracellular signal-regulated protein kinase †: extracellular signal-regulated protein kinase ‡: phosphorylated c-Jun amino-terminal kinases § : c-Jun amino-terminal kinases ∥ : phosphorylated P38 ¶ : inhibitory kappa Bα
그림 Fig. 5. Effects of Cheongpochukeo-tang (CCT) on the survival rate of mice. The mice were pre-intraperitoneal injected with CCT for 1 h, and then intraperitoneal injected with LPS (37.5 mg/kg). After 48 h, the survival rate of only-LPS-treated group was 0%. But after 120 h, the survival rate of CCT 0.1 was 20%, and the survival rate of CCT 0.2 was 50%. The number of mice of each group was 20.
그림 Fig. 6. Effects of Cheongpochukeo-tang (CCT) on the production of IL-1β, IL-6, and TNF-α in serum of mice. The serum of mice was pre-treated with CCT 0.01, 0.1, 0.2 (mg/kg) for 1 h, and then incubated with LPS (37.5 mg/kg) for 24 h. The level of cytokine was measured by ELISA. Data were given as means of values±SD from three independent experiments. * P<0.05 as compared with saline. †P<0.05 as compared with only-LPS-treated group.
그림 Fig. 7. Effects of Cheongpochukeo-tang (CCT) on the liver tissue of mice. Representative H & E stained sections of the liver tissue in the normal mice not given LPS, in the control mice given LPS, and in mice given CCT 1 h before the first LPS injections. Histological sections of the liver were scored from 0 (normal) to 3 (severe) for inflammation and hemorrhage. These figures show representative images of experiment group. The score was calculated by three pathologists who were blinded. * P<0.05 as compared with saline. †P<0.05 as compared with only-LPS-treated group.
그림 Fig. 8. Effects of Cheongpochukeo-tang (CCT) on the production of IL-1β, IL-6, and TNF-α in liver tissue of mice. The cells were pre-treated with CCT as indicated concentrations for 1 h, and then incubated with or without LPS (37.5 mg/kg) for 3 h. IL-1β, IL-6, and TNF-α levels were measured by real time RT-PCR. Data were given as means of values±SD from three independent experiments. * P<0.05 as compared with saline. †P<0.05 as compared with only-LPS-treated group.

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