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RISS 인기검색어
- 검색키워드
- 저자 : ZheMing Fang (검색결과 3 건)
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Effects of Red Deer Antlers on Cutaneous Wound Healing in Full-thickness Rat Models
Gu, LiJuan,Mo, EunKyoung,Yang, ZhiHong,Fang, ZheMing,Sun, BaiShen,Wang, ChunYan,Zhu, XueMei,Bao, JianFeng,Sung, ChangKeun Asian Australasian Association of Animal Productio 2008 Animal Bioscience Vol.21 No.2
The process of wound repair involves an ordered sequence of events such as overlapping biochemical and cellular events that, in the best of circumstances, result in the restoration of both the structural and functional integrity of the damaged tissue. An important event during wound healing is the contraction of newly formed connective tissues by fibroblasts. The polypeptide growth factors, like transforming growth factor-${\beta}$(TGF-${\beta}$, insulin-like growth factor I (IGF- I) and epidermal growth factor (EGF), play very important mediator roles in the process of wound contraction. Deer antlers, as models of mammalian regeneration, are cranial appendages that develop after birth as extensions of a permanent protuberance (pedicle) on the frontal bone. Antlers contain various growth factors which stimulate dermal fibroblast growth. They are involved in digestion and respiration and are necessary for normal wound healing and skin health. In order to investigate and evaluate the effects of red deer antlers on skin wound site, the speed of full-thickness skin wound healing and the expression of IGF-I, TGF-${\beta}$ and EGF in skin wounds, three groups of skin full-thickness rat models with a high concentration of antler ointment, a low concentration of antler ointment and without antler ointment were compared. At post-injury days 0, 2, 4, 8, 16, 20, 32, 40 and 60, the skin wound area was measured, the expressions of IGF-I, TGF- ${\beta}$ and EGF mRNA were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and collagen formation by sirius red dye and the localization of IGF-I, TGF-${\beta}$ and EGF peptides were inspected by histological immunohistochemical techniques. Wound healing was significantly more rapid in antler treated skins. In addition, the wound treated with a high concentration antler ointment, a low concentration antler ointment, and the control closed completely at post-injury day 40, day 44 and day 60, respectively. Via RT-PCR, the expressions of IGF-I (day 8 and day 16), TGF-${\beta}$(day 8, day 16 and day 20) and EGF (day 4, day 8, day 16, and day 32) were obviously up-regulated in high concentration antler-treated skins compared to control skins. Similar results could be seen in the histological detection of collagen dye and immunohistochemical methods using the corresponding polyclone antibodies of IGF-I, TGF-${\beta}$ and EGF. These results illustrate that antlers stimulate and accelerate the repair of cutaneous wounds.
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Partial Purification and Quantification of Insulin-like Growth Factor-I from Red Deer Antler
LiJuan Gu,Eunkyoung Mo(모은경),ZheMing Fang,BaiShen Sun,XueMei Zhu,Changkeun Sung(성창근) 한국생명과학회 2007 생명과학회지 Vol.17 No.10
사슴 뿔은 동물세계에서 가장 빨리 성장하는 조직이다. 따라서 성장중인 사슴 뿔은 뼈 성장을 촉진하는 인자가 풍부하게 포함된 것으로 생각된다. 이들 성장인자들 중 IGF-1은 뼈를 자라게 하는 조골세포의 대사에 중요한 역할을 한다고 알려져 있어 이를 정제하고자 하였다. IGF-1의 정제는 상대라고 불리는 신선한 사슴 뿔을 유안침전, DEAE-Sepharose CL-6B 이온교환수지, CM-Sepharose CL-6B 양이온교환수지, Sephadex G-50의 순차적인 방법으로 할 수 있었다. 각 과정마다 IGF-1의 거동을 HPLC, SDS-PAGE, Dot blot, 그리고 western blot으로 분석하였다. IGF-1의 정량은 ELISA기술로 재조합 인간 IGF-1을 이용하여 계산되었으며, 최종 분별 액은 두 개의 단백질을 보였으나, Western-blot에서 작은 분자량인 12 kDa으로 최종 판명할 수 있었다. 정제된 단백질은 HPLC에서 retention 시간 8분만에 검출되었으며, 총 농도는 2910 ng/㎖ 이고 중량은 0.291 g 이었다. Deer antler tissue contains the most rapidly growing bone in the animal kingdom. Thus, it is likely that growing antler tissue is a rich source of local paracrine bone-stimulating factors. Growth factors, at least the insulin-like growth factor (IGF), control the bone-remodelling process. In this study, we tried to isolate and purify IGF-I from fresh antler tissue by the routine isolation and purification of protein. The purification involved ammonium sulfate precipitation, DEAE-Sepharose CL-6B ion-exchange chromatography, CM-Sepharose CL-6B ion-exchange chromatography, and Sephadex G-50 chromatography. Purified fractions from each step were analyzed by high-performance liquid chromatography (HPLC), SDS polyacrylamide gel electrophoresis (SDS-PAGE), Dot-blot, and Western-blot methods. Furthermore, the quantification of partially purified IGF-I was calculated by enzyme-linked immunosorbent assays (ELISA) using antibody to human recombinant IGF-I. SDS-PAGE analysis of the final fraction yielded two molecular bands and the signal band was at 12 kDa on the Western-blot film. This purified IGF-I fraction showed a peak at retention time of eight min. The quantity of IGF-I in 20 g deer antler tissue as starting weight was calculated using a standard curve to be 2910 ng/㎖, and total IGF-I amount is 0.291 g. The results show that IGF-I, which can be found in deer antler, can be partially purified and quantified by classic protein isolation methods.
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Expression and localization of insulin-like growth factor-I in four parts of the red deer antler
Gu, Lijuan,Mo, Eunkyoung,Yang, Zhihong,Zhu, Xuemei,Fang, Zheming,Sun, Baishen,Wang, Chunyan,Bao, Jianfeng,Sung, Changkeun Informa Healthcare 2007 Growth factors Vol.25 No.4
<P> The expression and localization of insulin-like growth factor-I (IGF-I) in the four parts (tip, upper, mid and base) of the red deer antler has been extensively investigated. We used reverse transcriptase polymerase chain reaction (RT-PCR) and real-time reverse transcriptase polymerase chain reaction (real time RT-PCR), in situ hybridization, immunohistochemistry and Western blot techniques to localize IGF-I messenger ribonucleic acid (mRNA) and IGF-I peptide in the four parts of the antler. The specific sequence encoding IGF-I was detected by RT-PCR in all of the four specimens, and the 395 bp IGF-I sequence from the red deer antler was shown to have very high homology with human, goat and mouse IGF-I. In situ hybridization and immunohistochemistry results demonstrated that the expression of IGF-I occurred in chondrocytes and osteoblasts in the tip and upper parts of the antler. However, IGF-I was only detectable in osteoblasts around the bone in the mid and base parts. There were significant differences in the intensity of the signal obtained with the IGF-I probe in the tip, upper, mid and base tissues. The Western blot analysis also provided evidence that IGF-I expression was localized differentially in the four parts of the deer antler. This study indicates that antler tissue is an essential part of the IGF system, which is involved in the regulation of the growth of red deer antlers. The specific expression of IGF-I in the four parts of the deer antler suggests that the IGF-I molecule is present at significantly different levels throughout the deer antler development and regeneration processes. Localization of IGF-I in chondrocytes and osteoblasts suggests that IGF-I may play an important role in cartilage and bone formation. In addition, it may have a variety of biophysical effects that influence the rapid growth of deer antlers.</P>
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