Pharmacological effect of tetrahydroberberine from Corydalis Tuber on gastric motor dysfunction

다국어 초록 (Multilingual Abstract)

Functional dyspepsia is a condition associated with repetitive indigestion symptoms as early satiety, postprandial fullness or discomfort, vomiting and nausea in the central part of the epigastrium with no specific cause such as histopathological and biochemical organic lesion. Factors that have been proposed to explain functional dyspepsia symptoms are delayed gastric emptying, impaired gastric accommodation, visceral hypersensitivity, Helicobacter pylori infection and psychosocial effect. Drugs like prokinetic or fundus-relaxant are I clinical use and in development. However, there is currently no medication which shows efficacy of various causes of functional dyspepsia. It has a broad effect on gastrointestinal motility and sensory dysfunctions.
Tetrahydroberberine, an isoquinoline alkaloid from Corydalis Tuber, enhanced gastric emptying in an animal model. It showed the prokinetic property by stimulating increasing motor activity in the upper gastrointestinal including stomach, duodenum and jejunum. Also, it had a therapeutic effect on restraint stress-induced impaired gastric compliance, an experimental model that was established by this study. Moreover, it showed fundus-relaxing property by increasing meal-induced gastric accommodation in Beagle dogs.
The effects of tetrahydroberberine on gastrointestinal motor functions are mediated by serotonin 1A and 1B, dopamine 2, and adrenergic α2 receptors as well as nitric oxide. Based on DNA microarray, it changed the expression of stress-related neuropeptide. It had no effect on ataxia by its action on serotonin 1A receptor.
With these results, tetrahydroberberine may be a promising candidate in the control of functional gastrointestinal disorders including functional dyspepsia.

다국어 초록 (Multilingual Abstract)

기능성소화불량증 (functional dyspepsia)은 조직 병리적 및 생화학적인 기질적인 병변 등 특별한 원인 질환 없이 상복부 중앙에 조기 포만감, 만복감, 팽만감, 구역(또는 오심) 등의 소화 장애 증상이 반복되는 경우를 말한다. 그 원인으로는 위배출지연, 위순응장애, 과민내장, 헬리코박터균 감염, 사회심리적 영향 등이 제시되고 있다. 약물치료는 위장관운동촉진제 (prokinetics) 또는 기저부 이완제 (fundus-relaxant) 등이 임상적으로 사용되고 있거나 개발 중에 있다. 한편, 기능성 소화불량증의 다양한 병인에 대해 효력을 나타내거나 소화기 운동 및 감각 기능 장애에 대해 전반적으로 작용하는 치료제가 부재한 실정이다.
현호색 성분인 테트라히드로베르베린 (tetrahydroberberine)은 이소 퀴놀린계 알칼로이드 (isoquinolin alkaloid)로 동물모델에서 위배출을 촉진하였고 위장, 소장, 공장 등 상부위장관의 운동활성을 유의적으로 증가시키는 등 위장관 운동 촉진활성 (prokinetic property)을 나타내었다. 또한, 본 연구에서 확립한 구속스트레스 유발 위장 이완 장애에 대해 치료효과를 나타내었으며 각성 비글견의 식이 유발 위순응을 증가시키는 등 기저부 이완활성 (fundic-relaxing property)을 나타내었다.
이러한 소화관 운동기능에 대한 테트라히드로베르베린의 작용은 세로토닌 1A (5-HT1A) 및 1B (5-HT1B) 수용체, 도파민 2 (D2) 수용체, 아드레너직 알파 2 (α2 adrenergic) 수용체와 일산화질소(NO)을 매개로 하였으며 마이크로어레이 분석 결과, 스트레스 연관 뉴로펩타이드 (stress-related neuropeptide)의 발현 변화가 관찰되었다. 한편, 세로토닌 1A (5-HT1A) 수용체 작용에 의한 운동실조 (ataxia) 발현에 대한 영향은 없었다.
이러한 결과로부터 현호색 성분 테트라히드로베르베린은 기능성 소화불량증 등 기능성소화관장애를 치료하는데 있어 유용한 후보 물질이 될 수 있을 것으로 생각된다.

목차 (Table of Contents)

• I. Introduction ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙1
• Functional dyspepsia ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙1
• Pathophysiology ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙2
• Pharmacological approach∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙8
• Isoquinoline alkaloid for potential treatment of FD∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙12
• II. Materials and Methods ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙20
• 1. Materials ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙20
• 1.1. Plant material ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙20
• 1.2. Guidelines for experimental animals ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙20
• 1.3. Reagents ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙20
• 2. Methods ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙21
• 2.1. Extraction and isolation ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙21
• 2.2. Radioligand competition binding assays ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙22
• 2.2.1. Membrane preparation ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙22
• 2.2.2. Serotonin 5-HT1A receptor binding assay ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙22
• 2.2.3. Serotonin 5-HT3 receptor binding assay ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙22
• 2.2.4. Adrenergic alpha 2 receptor binding assay ∙∙∙∙∙∙∙∙∙∙∙∙∙∙23
• 2.2.5. Dopamine D2 receptor binding assay ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙23
• 2.2.6. Motilin receptor binding assay ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙24
• 2.2.7. Muscarinic M1, M2 or M3 receptor binding assay ∙∙∙∙24
• 2.2.8. Termination of binding assays and scintillation spectroscopy ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙24
• 2.2.9. Determination of other serotonin and ghrelin receptors binding activity ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙25
• 2.3. Gastric emptying ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙25
• 2.3.1. Animals and experimental procedure ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙25
• 2.3.2. Gastric emptying rate ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙26
• 2.4. Motor activity ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙26
• 2.4.1. Animals ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙26
• 2.4.2. Electromyography study ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙26
• 2.5. Restraint stress-induced feeding inhibition ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙27
• 2.5.1. Animals ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙27
• 2.5.2. Experimental procedure ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙27
• 2.5.3. Antagonist study ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙28
• 2.6. Impaired gastric compliance ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙28
• 2.6.1. Animals ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙28
• 2.6.2. Gastric barostat ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙29
• 2.6.3. Drug administration ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙30
• 2.6.4. Antagonist study ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙30
• 2.7. Canine gastric accommodation ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙31
• 2.7.1. Animals and experimental procedure ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙31
• 2.7.2. Antagonist study ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙32
• 2.8. DNA microarray analysis ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙32
• 2.8.1. Animals ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙32
• 2.8.2. Preparation of DNA probe and hybridization ∙∙∙∙∙∙∙∙∙∙∙∙32
• 2.8.3. Data analysis ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙33
• 2.9. Rotarod test ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙34
• III. Results ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙36
• 1. Isolation of tetrahydroberberine from Corydalis Tuber ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙36
• 2. In vitro activities of tetrahydroberberine ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙39
• 2.1. Radioligand competition binding assays ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙39
• 3. Prokinetic properties of tetrahydroberberine ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙41
• 3.1. Effect of tetrahydroberberine on gastric emptying in rats ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙41
• 3.2. Effect of tetrahydroberberine on motor activity in conscious rats ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙43
• 4. Fundus-relaxing properties of tetrahydroberberine ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙47
• 4.1. Effect of tetrahydroberberine on restraint stress-induced feeding inhibition in rats ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙47
• 4.2. Experimental animal model of impaired gastric compliance ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙50
• 4.3. Effect of tetrahydroberberine on impaired gastric compliance in restraint stress induced rats ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙52
• 4.4. Effect of tetrahydroberberine on canine gastric accommodation ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙55
• 5. Mode of action of tetrahydroberberine ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙61
• 5.1. Effect of NO synthase inhibitor and 5-HT1 receptor antagonists on tetrahydroberberine-induced rat stomach relaxation ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙61
• 5.2. DNA microarray analysis ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙64
• 6. Toxicity test of tetrahydroberberine ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙74
• 6.1. Rotarod test ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙74
• IV. Discussion ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙76
• V. Conclusion ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙82
• VI. References ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙84
• VII. Abstract ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙101
• VIII. Abstract in Korean ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙103