자하거 약침이 만성 신체 스트레스 모델 백서의 행동과 체중 변화, 수면 관련 호르몬, 염증성 사이토카인, 항산화 효소에 미치는 영향 :

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자료유형: 학위논문

저자정보: 하지원 (동의대학교, 동의대학교 대학원)

지도교수: 김보경

발행연도: 2019

저작권: 동의대학교 논문은 저작권에 의해 보호받습니다.

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2019

자하거 약침이 만성 신체 스트레스 모델 백서의 행동과 체중 변화, 수면 관련 호르몬, 염증성 사이토카인, 항산화 효소에 미치는 영향

하지원 한의학과 2019.01 학위논문

자하거 약침이 만성 신체 스트레스 모델 백서의 행동, 체중, 수면호르몬, 염증 사이토카인, 항산화 효소의 변화에 미치는 영향

하지원 , 이유미 , 나창수 외 2명 동의신경정신과학회지 2019.01 학술저널

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목적: 스트레스로 인한 불면에 대한 자하거 약침의 치료 효과를 확인하기 위해, 만성 신체 스트레스 모델을 적용한 백서를 대상으로, 자하거 약침 실험군과 대조군, 양성대조군 간의 행동 관련 지표, 체중 변화, 수면 관련 호르몬, 염증성 사이토카인, 항산화 효소, 혈액과 혈청 등을 비교 측정하였다.
방법: 300 g, 10 주령 이상의 Wistar rat을 6 개의 군으로 분리했다. 정상군에는 스트레스를 주지 않았고, 대조군과 양성대조군, 자하거 약침 실험군에는 만성 신체 스트레스 모델을 적용하였다. 대조군은 처치 않음, 양성대조군은 졸피뎀 투여, 자하거 약침 실험군(HPP군)에는 각각 0.5배 농도(HPP-0.5×군), 1배 원액(HPP-1×군), 2배량(HPP-2×군)의 자하거 약침을 주입했다. 모든 군의 Nestlet Shredding 행동 지표, 체중 변화, Melatonin, GABA, TNF-α, IL-6, SOD, GPx 수치, 혈액과 혈청을 비교 관찰했다.
결과: Nestlet Shredding은 정상군에 비해 대조군에서 유의한 증가를나타내었고(p<0.05), HPP-1×군과 HPP-2×군에서 대조군에 비해 유의하게 감소했다(p<0.05). 체중 변화는 2주 차와 3 주 차에 HPP-2×군과 양성대조군에서 대조군보다 체중 증가량이 더 많은 경향이 있었다. 수면 관련 호르몬에서 HPP-1×군과 HPP-2×군에서 대조군에 비해 GABA가 유의하게 증가했고(p<0.05) Melatonin 또한 증가 경향이 나타났다. TNF-α와 IL-6는 정상군에 비해 대조군에서 유의한 증가가 나타났고(p<0.01), 모든 자하거 실험군에서 대조군에 비해 유의하게 감소했다(p<0.05). SOD 함량은 정상군에 비해 대조군에서 유의한 감소가 나타났고(p<0.01), 대조군에 비해 모든 실험군에서 유의하게 증가하였다(p<0.05). GPx, AST, ALT, Bun, Creatinine과 백혈구 수치는 각 군간 유의한 차이가 없었다.
결론: 자하거 약침의 투여가 Nestlet Shredding 행동을 감소시키고 체중의 증가량을 늘리며, GABA 와 Melatonin, SOD를 증가시키고, TNF-α와 IL-6를 감소시키므로, 신체 스트레스를 완화하는 효과가 있고, 스트레스로 인한 수면장애의 치료에도 도움이 될 것으로 생각된다.

Aim: In this dissertation, the effect of Hominis Placenta Pharmacopuncture on behavior, sleep-related hormones, inflammatory cytokines, anti-oxidants, weight, blood and serum was investigated by designing the chronic physical stress rat model, to study the therapeutic value of Hominis Placenta Pharmacopuncture in treating insomnia caused by physical stress.
Methods: Wistar rats over 10 weeks of age were used. They were divided into 6 groups. Normal group was not subjected to any stress. Control group was subjected to chronic physical stress without any treatment. Positive control group was subjected to chronic physical stress and treated with zolpidem. 3 HPP(Hominis Placenta Pharmacopuncture) groups were subjected to chronic physical stress and treated with different densities of HPP; HPP-0.5×(0.5 times diluted), HPP-1×(undiluted) and HPP-2×(2 times condensed). Nestlet Shredding was performed, and GABA, Melatonin, TNF-α, IL-6, SOD, GPx, weight, AST, ALT, Bun, Creatinine and leukocytes were measured.
Results: Nestlet Shredding was increased in control group compared to the normal group(p<0.05), decreased in HPP-1× and HPP-2× compared to the control group(p<0.05). The change of weight showed increasing tendency in HPP-2× compared to the control group since the 2nd week. GABA was increased(p<0.05) and Melatonin also showed increasing tendency in HPP-1× and HPP-2× compared to the control group. TNF-α
and IL-6 were increased in control group compared to the normal group(p<0.01), and decreased in all HPPs compared to the control group(p<0.05). SOD level was decreased in control group compared to the normal group(p<0.01), and increased in all HPPs compared to the control group(p<0.05). GPx, AST, ALT, Bun, Creatinine and leukocytes showed no noticeable difference among all groups.
Conclusion: Hominis Placenta Pharmacopuncture had affected on increasing weight, GABA, Melatonin, SOD, and decreasing Nestlet Shredding and inflammatory cytokines, suggesting that it helps relieve physical stress, and treat insomnia consequently.

I. INTRODUCTION ······································································································ 1
II. MATERIALS AND METHODS ·········································································· 3
1. Animals ··············································································································· 3
2. Pharmacopuncture and Drug Treatment ··················································· 3
3. CPS(Chronic Physical Stress) Procedure ·················································· 5
4. Nestlet Shredding ··························································································· 7
5. Body Weight ····································································································· 7
6. ELISA(Enzyme Linked Immunosorbent Assay) Test for Melatonin, GABA, TNF-α and IL-6 7
6.1. Melatonin ······························································································ 7
6.2. GABA(Gamma-Aminobutylic Acid) ··················································· 8
6.3. TNF-α(Tumor Necrosis Factor Alpha) ············································ 8
6.4. IL-6(Interleukin-6) ············································································· 9
7. RT-PCR(Reverse Transcription Polymerase Chain Reaction) for SOD and GPx 9
7.1. Separation of Total RNA ································································· 9
7.2. RT-PCR ··································································································· 10
8. Analysis of Blood and Serum ·································································· 11
9. Statistical Analysis ······················································································ 11
III. RESULTS ················································································································ 12
1. Effect on the Behavior of Nestlet Shredding ······································ 12
2. Effect on the Weight Change ···································································· 13
3. Effect on the Sleep-related Hormones ··················································· 14
3.1. Melatonin ··································································································· 14
3.2. GABA(Gamma-aminobutylic Acid) ···················································· 15
4. Effect on Inflammatory Cytokines ···························································· 16
4.1. TNF-α(Tumor Necrosis Factor Alpha) ············································ 16
4.2. IL-6(Interleukin-6) ·················································································· 17
5. Effect on Anti-oxydant Enzymes ····························································· 18
5.1. SOD(Superoxide dismatase) ································································· 18
5.2. GPx(Glutathione peroxidase) ································································ 19
6. Effect on Blood and Serum ········································································ 20
6.1. AST, ALT, Bun, and Creatinine ························································ 20
6.2. Leukocytes ································································································ 21
IV. DISCUSSION ·································································································· 23
V. CONCLUSION ································································································· 31
REFERENCES ······························································································· 33
Abstract ··················································································································· 41

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