[논문]진피-황금 혼합물이 급성 역류성 식도염 흰쥐에 미치는 효과

이진아; 신미래; 노성수; 박해진

doi:10.4163/jnh.2021.54.3.321

진피-황금 혼합물이 급성 역류성 식도염 흰쥐에 미치는 효과
Effects of a mixture of Citri Pericarpium and Scutellariae Radix on acute reflux esophagitis in rats 원문보기

Journal of nutrition and health, v.54 no.3, 2021년, pp.321 - 333

이진아 (대구한의대학교 한의과대학 본초약리학교실) , 신미래 (대구한의대학교 한의과대학 본초약리학교실) , 노성수 (대구한의대학교 한의과대학 본초약리학교실) , 박해진 (대구한의대학교 DHU 바이오융복합시험센터)

초록
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본 연구에서는 진피-황금 혼합물 (CS)이 급성 역류성 식도염에 미치는 식도 점막 보호 효과를 평가하기 위하여 CS를 경구투여한 후 수술을 통해 역류성 식도염을 유발하였으며, 실험 종료 후 혈액 채취 및 식도 조직을 적출하였다. 동물에게서 적출한 식도 점막의 손상 정도를 육안으로 확인한 결과 CS투여군에서 식도 점막의 손상이 유의하게 감소하였으며, H&E staining을 통해 관찰한 결과 마찬가지로 CS투여군에서 식도 상피의 탈락 및 염증세포의 침윤이 현저하게 감소한 것을 확인하였다. 혈액을 이용하여 역류성 식도염의 원인으로 유효하다고 알려진 ROS의 수치를 확인한 결과, CS투여군에서 ROS 수치가 유의적으로 감소하였으며, western blotting을 통해 NADPH oxidase인 NOX4, p47^phox, p22^phox의 발현을 확인한 결과, 마찬가지로 CS 투여군에서 유의하게 감소하였고, 특히 CS200투여군에서 Normal군과 비슷한 수치를 나타냈다. 또한, CS투여는 염증성 단백질인 MAPK와 NF-κB 경로를 유의적으로 억제하였을 뿐 아니라 tight junction 단백질인 claudin-1과 claudin-4의 발현을 유의하게 조절한 것을 확인하였다. 이상의 결과를 종합해보면 진피-황금 추출물은 산화적 스트레스를 억제함으로써 염증성 단백질의 발현을 조절할 뿐 아니라 tight junction 단백질의 발현을 조절하여 식도 점막을 보호하는 것으로 판단되나 역류성 식도염은 음식물의 섭취와 밀접한 관련이 있는 만큼 추후 동물의 식이 섭취량을 조사하는 등 세부적인 추가 연구가 필요할 것으로 보인다.

Abstract ▼ AI-Helper

Purpose: Reflux esophagitis is a disease caused by the reflux of stomach contents and stomach acid etc. into the esophagus due to defect in the lower esophageal sphincter and is currently increasing worldwide. This study was conducted to evaluate the effect of a mixture of Citrus Reticulata and Scutellariae Radix (CS) extract on acute reflux esophagitis in rats. Methods: Rats were divided into five groups for examination: normal group (Normal, n = 8), water-treated acute reflux esophagitis rats (Control, n = 8), tocopherol 30 mg/kg body weight-treated acute reflux esophagitis rats (Toco, n = 8), CS 100 mg/kg body weight-treated acute reflux esophagitis rats (CS100, n = 8), CS 200 mg/kg body weight-treated acute reflux esophagitis rats (CS200, n = 8). The experimental groups were administrated of each treatment compounds and after 90 min, acute reflux esophagitis was induced through surgery. Rats were sacrificed 5 h after surgery. We measured the level of reactive oxygen species (ROS) in serum and analyzed the expression of nicotinamide adenine dinucleotide phosphate, inflammatory, and tight junction-related proteins by western blot in the esophageal tissues. Results: CS administration significantly protected the esophageal mucosal damage due to reflux esophagitis, and the level of ROS in the serum was significantly reduced with CS administration as compared to Control. In addition, CS administration significantly suppressed mitogen-activated protein kinase (MAPK or MAP kinase) and nuclear factor-kappa B (NF-κB) pathways and increased protein expressions of tight junction protein. Conclusion: These results suggest that the CS not only regulates the expression of inflammatory proteins by inhibiting oxidative stress, but also reduces damage to the esophageal mucosa by inhibiting the expression of tight junction proteins.

주제어

표/그림 (8)

표 Table 1. Nutritional component of NIH-41 diet
그림 Fig. 1. Surgical induction of acute reflux esophagitis. A representative gross image; (A), esophageal ulcer ratio; (B). Esophageal ulcer ratio = damaged area of esophagus/total area of esophagus × 100. All data are expressed means ± SD (n = 8). Normal, normal rats; Control, acute reflux esophagitis rats; Toco, tocopherol 30 mg/kg body weight/day-treated acute reflux esophagitis rats; CS100, mixture of Citrus Reticulata and Scutellariae Radix 100 mg/kg body weight/day-treated acute reflux esophagitis rats; CS200, mixture of Citrus Reticulata and Scutellariae Radix 200 mg/kg body weight/day-treated acute reflux esophagitis rats. Significance: ^###p < 0.001 vs. Normal rats and **p < 0.01, ***p < 0.001 vs. Control rats.
그림 Fig. 2. Esophagus histological examination through hematoxylin and eosin staining (magnification ×200). Normal, normal rats; Control, acute reflux esophagitis rats; Toco, tocopherol 30 mg/kg body weight/day-treated acute reflux esophagitis rats; CS100, mixture of Citrus Reticulata and Scutellariae Radix 100 mg/kg body weight/day-treated acute reflux esophagitis rats; CS200, mixture of Citrus Reticulata and Scutellariae Radix 200 mg/kg body weight/day-treated acute reflux esophagitis rats.
그림 Fig. 3. ROS level in serum. ROS level = Absorbance_30min -Absorbance_0min/30 × 200. All data are expressed means ± SD (n = 8). ROS, reactive oxygen species; Normal, normal rats; Control, acute reflux esophagitis rats; Toco, tocopherol 30 mg/kg body weight/day-treated acute reflux esophagitis rats; CS100, mixture of Citrus Reticulata and Scutellariae Radix 100 mg/kg body weight/day-treated acute reflux esophagitis rats; CS200, mixture of Citrus Reticulata and Scutellariae Radix 200 mg/kg body weight/day-treated acute reflux esophagitis rats. Significance: ^###p < 0.001 vs. Normal rats and **p < 0.01, ***p < 0.001 vs. Control rats.
그림 Fig. 4. Expression of NADPH oxidase proteins in esophagus. Representative images of NOX4, p47^phox, p22^phox, and β-actin proteins (A). The expression level of NOX4 (B), p47^phox (C), p22^phox (D) were quantified. All data are expressed mean ± SD (n = 8). Normal, normal rats; Control, acute reflux esophagitis rats; Toco, tocopherol 30 mg/kg body weight/day-treated acute reflux esophagitis rats; CS100, mixture of Citrus Reticulata and Scutellariae Radix 100 mg/kg body weight/day-treated acute reflux esophagitis rats; CS200, mixture of Citrus Reticulata and Scutellariae Radix 200 mg/kg body weight/day-treated acute reflux esophagitis rats. Significance: ^###p < 0.001 vs. Normal rats and *p < 0.05, **p < 0.01, ***p < 0.001 vs. Control rats.
그림 Fig. 5. Expression of MAPK proteins in esophagus. Representative images of c-Fos, c-Jun, p-p38, p-ERK, P-JNK, and β-actin proteins (A). The expression level of c-Fos (B), c-Jun (C), p-p38 (D), p-ERK (E), p-JNK (F) were quantified. All data are expressed means ± SD (n = 8). Normal, normal rats; Control, acute reflux esophagitis rats; Toco, tocopherol 30 mg/kg body weight/day-treated acute reflux esophagitis rats; CS100, mixture of Citrus Reticulata and Scutellariae Radix 100 mg/kg body weight/day-treated acute reflux esophagitis rats; CS200, mixture of Citrus Reticulata and Scutellariae Radix 200 mg/kg body weight/day-treated acute reflux esophagitis rats. Significance: ^##p < 0.01, ^###p < 0.001 vs. Normal rats and *p < 0.05, **p < 0.01, ***p < 0.001 vs. Control rats.
그림 Fig. 6. Expression of inflammation related proteins in esophagus. Representative images of NF-κBp65, p-IκBα, COX-2, TNF-α, IL-6, and β-actin proteins (A). The expression level of NF-κBp65 (B), p-IκBα (C), COX-2 (D), TNF-α (E), IL-6 (F) were quantified. Normal, normal rats; Control, acute reflux esophagitis rats; Toco, tocopherol 30 mg/kg body weight/day-treated acute reflux esophagitis rats; CS100, mixture of Citrus Reticulata and Scutellariae Radix 100 mg/kg body weight/day-treated acute reflux esophagitis rats; CS200, mixture of Citrus Reticulata and Scutellariae Radix 200 mg/kg body weight/day-treated acute reflux esophagitis rats. Significance: ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 vs. Normal rats and *p < 0.05, **p < 0.01, ***p < 0.001 vs. Control rats.
그림 Fig. 7. Expression of tight junction proteins in esophagus. Representative images of Claudin-1, Claudin-4, and β-actin proteins (A). The expression level of Claudin-1 (B) and Claudin-4 (C) were quantified. All data are expressed means ± SD (n = 8). Normal, normal rats; Control, acute reflux esophagitis rats; Toco, tocopherol 30 mg/kg body weight/day-treated acute reflux esophagitis rats; CS100, mixture of Citrus Reticulata and Scutellariae Radix 100 mg/kg body weight/day-treated acute reflux esophagitis rats; CS200, mixture of Citrus Reticulata and Scutellariae Radix 200 mg/kg body weight/day-treated acute reflux esophagitis rats. Significance: ^##p < 0.01 vs. Normal rats and *p < 0.05, **p < 0.01 vs. Control rats.

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