[보고서]당뇨병성 말초신경병증 치료 한약제제 BJT의 본초 효능별 비교 탐색연구

조충식

당뇨병성 말초신경병증 치료 한약제제 BJT의 본초 효능별 비교 탐색연구 원문보기

보고서 정보
주관연구기관	대전대학교 산학협력단 Daejeon University
연구책임자	조충식
보고서유형	최종보고서
발행국가	대한민국
언어	한국어
발행년월	2016-06
과제시작연도	2015
주관부처	보건복지부 [Ministry of Health & Welfare(MW)(MW)
등록번호	TRKO201700000708
과제고유번호	1465017953
사업명	한의약선도기술개발사업
DB 구축일자	2017-09-20
키워드	당뇨병.당뇨병성 말초신경병증.보기제통탕.효능.본초.diabetes mellitus.diabetic neuropathy.BJT.effectiveness.medical herbs.

초록 ▼

1. Taxol과 신경손상에 의하여 유도된 말조신경의 병증적 반응성에 대해 BJT와 변증별 분류 그룹에서 신경병증성 반응성의 개선 효과를 나타냄을 확인함.
- 신경병증조직의 슈반세포의 보호효과
- 감각신경세포의 신경돌기성장의 개선효과
- in vivo 조직의 병증반응성 개선, 손상신경의 축삭재생의 개선 관찰
- 행동학적 개선
- Be 그룹에서 보기제통탕과 상당수준 유사한 효능성을 나타냄을 확인함
2. STZ의 주입에 의하여 유도된 당뇨병성 동물모델에 마찬가지로 BJT와 변증별 분류 약물의 처리시 유사한 방식의 양성적 효능성을 관찰함
- 당수치개선 및 염증반응성 개선
- 세포생존 지표단백질(p-Erk1/2)의 생성증가 및 자멸경로 단백질(p38) 생성수준의 감소
- 감각신경세포의 신경돌기 성장 개선
- 감각신경의 행동학적 효능성 개선
3. HPLC 분석을 통한 지표물질 확보함

(출처 : 보고서 요약서)

Abstract ▼

Results
1. Examination of the protective effects of BJT on the peripheral nerve from damage caused by taxol and nerve crush
BJT treatment enhanced neurite outgrowth in cultured DRG neurons and improved Schwann cell survival. Phospho-Erk1/2 levels were elevated by BJT treatment in the injured- or taxol-treated sciatic nerves. In the injured nerve, BJT improved regenerative responses when measured by biochemical and histological analyses. A drug group (Be) composed of 4 active herbal components which were selected by neurite growth-enhancing activity was as effective as BJT for the recovery of thermal sensitivity of the hind paws which had been suppressed by taxol administration.

2. Examination of protective effects of BJT and its selected formula on neuropathic insults in streptozotocin-induced diabetic animals
Be decoction was formulated by selecting individual herbal components that induced neurite outgrowth most efficiently in each subgroup. Be decoction was most effective in inducing the neurite outgrowth among subgroup decoctions in DRG neurons that had been prepared from STZ-diabetic animals. Both BJT and Be decoction downregulated the production of TNF-α from the sciatic nerves in STZdiabetic animals. While levels of phospho-Erk1/2 in the sciatic nerves of STZ-treated animals were increased by BJT and Be treatments, levels of p38 kinase were decreased by both drug treatments. Oral administration of BJT and Be in STZ-diabetic animals reduced the latency time responding to thermal stimulation in the hot plate test.

(출처 : SUMMARY)

목차 Contents

표지 ... 1제 출 문 ... 2보고서 요약서 ... 3국문 요약문 ... 5SUMMARY ... 6목차 ... 71. 연구개발과제의 개요 ... 8 1-1. 연구개발 목적 ... 8 1-2. 연구개발의 필요성 ... 8 1-3. 연구개발 범위 ... 112. 국내외 기술개발 현황 ... 133. 연구수행 내용 및 결과 ... 164. 목표달성도 및 관련분야 기여도 ... 31 4-1. 목표달성도 ... 31 4-2. 관련분야 기여도 ... 315. 연구결과의 활용계획 ... 326. 연구과정에서 수집한 해외과학기술정보 ... 337. 연구개발성과의 보안등급 ... 338. 국가과학기술종합정보시스템에 등록한 연구시설·장비 현황 ... 339. 연구개발과제 수행에 따른 연구실 등의 안전조치 이행실적 ... 3410. 연구개발과제의 대표적 연구실적 ... 3511. 기타사항 ... 3512. 참고문헌 ... 36끝페이지 ... 38

표/그림 (14)

표 Sources of BJT
표 Morphological changes of sciatic nerve axons and DRG neurons after taxol injection and BGJTD treatments. Sciatic nerve was exposed and treated with DMSO vehicle, taxol, or taxol plus BGJTD, and longitudinal sections of the nerve at the injectedlocation (A,C) and DRG at lumbar level 5 (D) were used for immunofluorescence staining for NF-200 and βIII-tubulin.The number of axons whose length is longer than 100㎛, as illustrated with dotted rectangles in (A), wascounted from the images, averaged from 3 nonconsecutive sections and compared among 3 experimental groups. Quantitative data are shown in (B). Soma DRG neurons wasmarked by arrows in (D).
표 Effects of BGJTD on the neurite outgrowth of DRG neurons.
표 Regulation of Schwann cell survival by BGJTD treatment. Schwann cells prepared from intact sciatic nerve were treated with DMSO vehicle, taxol, and taxol plus BGJTD. Fixed cells were used for immunostaining with anti-S100β antibody (A)and anti-caspase 3 antibody (B). Cells were also stained with Hoechst33258 to visualize individual nuclei (blue). (A) Morphological comparison of S100β-stained Schwann cells after different treatments. (B) Immunofluorescence staining of caspase 3 in cultured Schwann cells. (C) Quantitative comparison of caspase 3-positive cells in cultured cells after different treatments. A percentage of caspase 3-positive cellsfrom randomly selected microscopic fields (more than 5 fields) were determined from each experiment
표 Effects of taxol and BGJTD treatments on Erk1/2 and Cdc2 activity in the sciatic nerve.Western blot analysis of phospho-Erk1/2 in the sciatic nerves (A) and in the DRG (B) after taxol injection and BGJTD treatments (T+B).(C) Western blot analysis of Cdc2 and phospho-vimentin in the sciatic nerves treated with DMSO vehicle, taxol, and taxol plus BGJTD.(D) Representative images showing Cdc2 and phospho-vimentin signals in S100β-labeled Schwann cells. Longitudinal sections from distal portion of the nerve treated with taxol and BGJTD were used in this experiment. Western blotting images in (A-C) are the representatives from 3 independent experiments. Quantitative comparison of mean band intensity for target proteins relative to actin control in western blotting are shown in lower panels in (A-C). Scale bars in (D): 100 ㎛.
표 Effects of BGJTD treatment on the axonal regeneration after nerve injury
표 Effects of BGJT subgroup decoctions on neurite outgrowth of taxol-treated DRG sensory neurons.Six days after taxol or DMSO vehicle injection in vivo into the sciatic nerve, DRG neurons were cultured in the presence of individual decoctionsBa to Be (0.5 mg/ml) for 48 hr and harvested for immunostaining. (A) Comparison of neurite length among experimental groups. (B) Representative images of cultured cells after immunofluorescence staining with NF-200 (Green) and GAP-43 (red). Cell nuclei were visualized by nuclear staining with Hoechst 33258 (blue). In (A), neurite length was determined by analyzing more than 20 neurons in 7-10 random microscopic fields, averaged 4 independent experiments, and compared among experimental groups. Error bars in (A) denote standard error of mean (SEM).
표 Effects of taxol, BGJTD, and Be drugs on the heat sensitivity. Mice were given taxolfollowed by oral administration of BGJTD or Be, and were subjected to measure the lifting frequency of the hind paws (A) and the latency period after placing on the hot plate (B). Error bars in (A) denote standard error of mean (SEM).
표 Comparison of neurite outgrowth in DRG neurons after the treatment of individual constituents of BGJTD. Herbal drugs (0.5 mg/ml) were treated to DRG neurons for 48hr and harvested for immunofluorescence staining for NF-200 (green) and GAP-43 (red) signals
표 Effects of the treatment of subgroup decoctions on blood glucose levels, TNF-α production and neurite outgrowth
표 Comparison of TNF-α levels in the sciatic nerves of STZ-diabetic animals after different treatments. (CTL; untreated control, SAL; saline injection). Animals were treated with herbal decoctions for 2 weeks after 1 week of STZ injection
표 Induction pattern of phospho-Erk1/2 and p38 proteins in the sciatic nerves of STZ-induced diabetic animals after different treatments. (CTL; untreated control, SAL; saline injection).
표 Effects of BGJTD and Be decoction on the heat sensitivity. STZ-induced diabetic mice were orally administered BGJTD and Be for 2 weeks, and were subjected to the measurement of the lifting frequency of the hind paws (A) and the latency period after placing on the hot plate (B). Error bars in (A) denote SEM.
표 HPLC profile of BGJTD and its subgroup extracts.Peaks that coincide with standard chemical solutions are marked by vertical arrows, and UV values applied to detect peaks are indicated in the figure.

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당뇨병성 말초신경병증 치료 한약제제 BJT의 본초 효능별 비교 탐색연구 원문보기