[논문]고지방식이로 유도된 대사증후군 모델 동물에서 백호가인삼탕(白虎加人參湯)의 장내미생물 및 유전자 발현 조절을 통한 대사 개선 효과

조민진; 한송이; 임수경; 송은지; 남영도; 김호준

doi:10.18325/jkmr.2023.33.3.1

[국내논문] 고지방식이로 유도된 대사증후군 모델 동물에서 백호가인삼탕(白虎加人參湯)의 장내미생물 및 유전자 발현 조절을 통한 대사 개선 효과
The Effect of Baekhogainsam-tang on Metabolism through Modulation of the Gut Microbiota and Gene Expression in High-Fat Diet Induced Metabolic Syndrome Animal Model 원문보기

Journal of Korean Medicine Rehabilitation : JKMR = 한방재활의학과학회지, v.33 no.3, 2023년, pp.1 - 15

조민진 (동국대학교 한의과대학 한방재활의학교실) , 한송이 (동국대학교 한의과대학 한방재활의학교실) , 임수경 (동국대학교 한의과대학 한방재활의학교실) , 송은지 (한국식품연구원 헬스케어연구단) , 남영도 (한국식품연구원 헬스케어연구단) , 김호준 (동국대학교 한의과대학 한방재활의학교실)

Abstract ▼ AI-Helper

Objectives We aimed to find out the improvement effect of Baekhogainsam-tang (Baihu Jia Renshen-tang, BIT) on metabolic syndrome and alteration of microbiota and gene expression. Methods We used male C57BI/6 mice and randomly assigned them into three groups. Normal control group was fed 10% kcal% fat diet, high-fat diet (HFD) group was fed 45% kcal% fat diet and 10% fructose water. BIT group was fed same diet as HFD group and treated by BIT for once daily, 6 days per week, total 8 weeks. We measured their body weight and food intake every week and performed oral glucose tolerance test 1 week before the end of the study. Then we collected the blood sample to measure triglyceride, total cholesterol, high-density lipoprotein cholesterol, insulin, and hemoglobin A1c. We harvested tissue of liver, muscle, fat, and large intestine for quantitative polymerase chain reaction (qPCR) and histopathological examination. Fresh fecal samples were collected from each animal to verify alterations of gut microbiota and we used RNA from liver tissue for microarray analysis. Results The body weight and fat weight of BIT group were reduced compared to HFD group. The qPCR markers usually up-regulated in metabolic syndrome were decreased in BIT group. Bacteroides were higher in BIT group than other groups. There were also differences in gene expressions between two groups such as Cyp3a11 and Scd1. Conclusions We could find out BIT can ameliorate metabolic syndrome and suggest its effect is related to gut microbiota composition and gene expression pattern.

Keyword

표/그림 (9)

그림 Fig. 1. The results of kcal intake, body weight, liver weight and fat mass. (A) The amount of kcal intake and its weekly changes among three groups in 2 weeks of pretreatment (p1, p2) and 8 weeks of treatment. (B) The body weight of three groups and its weekly changes were monitored during the same period. (C) The weight of liver weight and fat mass (total fat, subcutaneous fat, mesenteric fat and epididymal fat) of mice were measured. All data represents standard error of mean. NC: normal control, HFD: high-fat diet, BIT: Baekhogainsam-tang, p1: first week of pretreatment, p2: second week of pretreatment. ^##p<0.01, ^###p<0.001, ^####p<0.0001 (#: vs. NC). *p<0.05, **p<0.01 (*: vs. HFD).
그림 Fig. 2. The effect of BIT on glucose and lipid level in serum. (A) The results of glucose AUC of glucose tolerance test among three groups 1 week before the end of the experiment. The changes of blood glucose level for 120 minutes were also monitored. From the blood sample of mice, (B) insulin, (C) HbA1c, (D) triglyceride, (E) total cholesterol, and (F) HDL cholesterol of three groups were measured after 8 weeks of treatment. All data represents standard error of mean. AUC: area under the curve, OGTT: oral glucose tolerance test, NC: normal control, HFD: high-fat diet, BIT: Baekhogainsam-tang, HbA1c: hemoglobin A1c, HDL: high-density lipoprotein cholesterol. ^#p<0.05, ^##p<0.01, ^####p<0.0001 (#: vs. NC). **p<0.01, ***p<0.001, ****p<0.0001 (*: vs. HFD).
그림 Fig. 3. The results of liver mRNA expression by quantitative polymerase chain reaction and histopathological findings of liver tissue in each group. (A) Genes analyzed from liver tissue are related to gluconeogenesis, lipogenesis, and inflammation. (B~D) Samples were done by hematoxylin and eosin staining and white circles manifest the lipid droplets (×200); These are (B) the result of NC group, (C) HFD group, and (D) BIT group. (E~G) Samples were done by Oil Red O staining and the red sites manifest the distribution of lipid (×400); These are (E) the result of NC group, (F) HFD group, and (G) BIT group. All data represents standard error of mean. NC: normal control, HFD: highfat diet, BIT: Baekhogainsam-tang, PEPCK: phosphoenolpyruvate carboxykinase, GLUT2: glucose transporter type 2, G6Pase: glucose-6-phosphatase, LPL: lipoprotein lipase, PPAR-γ: peroxisome proliferator-activated receptor γ, ACC1: acetyl-CoA carboxylase 1, TNF-α: tumor necrosis factor-α, MCP-1: monocyte chemoattractant protein-1. ^#p<0.05, ^##p<0.01 (#: vs. NC). *p<0.05, **p<0.01 (*: vs. HFD).
그림 Fig. 4. The results of skeletal muscle mRNA expression by quantitative polymerase chain reaction in each group related to glucose metabolism, fatty acid uptake, and mitochondrial biogenesis. All data represents standard error of mean. NC: normal control, HFD: high-fat diet, BIT: Baekhogainsam-tang, GLUT4: glucose transporter type 4, CPT-1α: carnitine palmitoyltransferase I, NRF1: nuclear respiratory factor 1, PGC1α: peroxisome proliferatoractivated receptor γ coactivator 1-α. ^##p<0.01, ^###p<0.001, ^####p< 0.0001 (#: vs. NC). *p<0.05, ***p<0.001, ****p<0.0001 (*: vs. HFD).
그림 Fig. 5. The results of epididymal white adipose tissue (eWAT) mRNA expression by quantitative polymerase chain reaction and histopathological findings of white adipose tissue in each group. (A) Genes are related to adipokine and lipogenesis. (B~D) Samples were done by hematoxylin and eosin staining (×200). (B) It represents the size of the adipocyte area in NC group, (C) represents HFD group, and (D) represents BIT group. (E) It is the statistical results comparing the size of the adipocyte area among three groups. NC: normal control, HFD: high-fat diet, BIT: Baekhogainsam-tang, LPL: lipoprotein lipase, PPAR-γ: peroxisome proliferator-activated receptor γ. ^#p<0.05, ^##p<0.01, ^###p<0.001, ^####p<0.0001 (#: vs. NC). *p<0.05, **p<0.01, ****p<0.0001 (*: vs. HFD).
그림 Fig. 6. The results of large intestine mRNA expression by quantitative polymerase chain reaction and histopathological findings of large intestine tissue in each group. (A) Genes are related to gut permeability and inflammation. (B~D) The large intestine tissue was stained with periodic acid-Schiff staining (×400). The villi of the intestine was stained purple; (B) It is the intestinal state of NC group, (C) HFD group, and (D) BIT group. All data represents standard error of mean. NC: normal control, HFD: high-fat diet, BIT: Baekhogainsam-tang, ZO-1: zonula occludens-1, IL-1β: interleukin-1 beta, MCP-1: monocyte chemoattractant protein-1, TLR2: toll-like receptor 2. ^#p<0.05, ^##p<0.01, ^####p<0.0001 (#: vs. NC). **p<0.01, ***p< 0.001, ****p<0.0001 (*: vs. HFD).
그림 Fig. 7. Diversity and abundance of gut microbiota in each group. (A) It was analyzed by Shannon index and (B) it was analyzed by Chao1 index for α-diversity. (C) The distances of each group were represented by β-diversity and it was performed by using principal coordinates analysis. (D) Altered community composition of gut microbiota and relative taxa abundance among three groups at the genus level. "Not_Assigned" stands for the sequence reads not included at genus level. The data for α-diversity presents mean±interquartile range. Relative abundance represents as percentage. NC: normal control, HFD: high-fat diet, BIT: Baekhogainsam-tang.
그림 Fig. 8. Scatter plot and heatmap of altered gene expression. (A) Scatter plot represents comparison of gene expression between BIT group and HFD group. The red dots placed at upper side of red dotted line represent up-regulated gene in BIT group compared to HFD group. The green dots placed at lower side of green dotted line represent down-regulated gene in BIT group. (B) Heatmap shows the change of gene expression. Left side indicates increased/decreased gene at HFD group compared to NC group. Right side indicates increased/decreased gene at BIT group compared to HFD group. The color red represents up-regulated gene and the color purple represents down-regulated gene. NC: normal control, HFD: high-fat diet, BIT: Baekhogainsam-tang.
그림 Fig. 8. Continued.

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저자의 다른 논문 :

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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