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중증하지허혈 모델에서 골수유래 중간엽줄기세포가 혈관 형성에 미치는 영향
손명주(Myeongjoo Son),강웅철(Woong Chol Kang),변경희(Kyunghee Byun) 대한체질인류학회 2016 해부·생물인류학 (Anat Biol Anthropol) Vol.29 No.2
중증하지허혈 (Critical limb ischemia, CLI)은 동맥경화성 말초동맥질환의 가장 심한 임상 양상 중 하나로 혈전에 의해 유발되어 하지혈관을 통한 혈류를 감소시키고 피부궤양이나 조직괴사, 진행 정도에 따라 사지 절단을 초래할 수 있다. 질환 치료를 위해서 경피적 혈관 중재술 (percutaneous peripheral intervention) 등을 시도하고 있으나 이는 혈관을 뚫어줄 뿐 혈관 재협착률을 줄이지 못하며 새로운 혈관 생성을 하지 못하기 때문에 사지 절단률과 사망률을 유의하게 줄이지 못하고 있다. 최근 골수유래 줄기세포의 주변분비효능 (paracrine effect)을 통한 허혈성 질환에서의 치료 효과에 대해서 보고되었다. 본 연구에서는 중증하지허혈 동물 모델에서 골수유래 중간엽줄기세포의 치료 및 신혈관 형성 (angiogenesis) 효능을 확인하고자 한다. 이를 위해 사람의 골수에서 분리한 중간엽줄기세포의 특성을 유세포분석과 분화실험을 통해 확인하였다. 사람제 대정맥혈관내피세포에서 줄기세포의 배양 배지를 처리한 후 혈관 형성 확인을 통해 주변분비효능을 확인하였다. 또한 10% Ferric chloride (FeCl3)를 이용하여 흰쥐의 넙다리동맥 내에 혈전을 생성하여 중증하지허혈과 비슷한 조건을 만들었으며, 10만 개의 사람골수유래 중간엽줄기세포를 넙다리동맥의 주행에 따라 다리근육에 주입하였다. 28일 후에 줄기세포의 생착능, 다리혈관의 변화, 다리근육의 상태와 섬유화 등을 비교하여 주입한 줄기세포의 효능을 확인하였다. 실험 결과 줄기세포 배양 배지 처리 후 혈관내피세포의 혈관 형성능이 대조군보다 높았을 뿐만 아니라 주입한 중 간엽줄기세포를 주입한 흰쥐에서의 신생혈관 형성능이 높았다. 다리의 상태를 육안으로 확인한 허혈 정도 또한 좋아졌으며, 다리근육 세포의 조직학적 상태와 섬유화 정도에서도 유의미하게 호전되었다. 이상의 실험 결과를 통해 골수에서 분리 배양한 중간엽줄기세포는 중증하지허혈에서의 혈관 형성능뿐만 아니라 섬유화 억제 등의 효과로 중증하지허혈에서 하지 절단을 예방할 수 있음을 확인하였다. Critical limb ischemia (CLI) is the most severe peripheral artery disease and caused by thrombus formation in blood vessel. The current strategies for treating CLI does not protect limb amputation and reduction in the risk of mortality. Recently, human bone marrow derived mesenchymal stem cells (BD-MSC) were reported to have a paracrine effects on angiogenesis in several ischemic diseases. So, we validate to determine whether BD-MSC protect against ferric chloride treated CLI and induce angiogenesis. To characterized human bone marrow derived stem cell, BD-MSC differentiated to osteocytes and adipocytes and validated stemness using flow cytometry. Endothelial cell induced angiogenesis followed by mesenchymal stem cell cultured medium treatment in HUVEC in vitro. We also mimicked CLI patients condition using FeCl3 treated CLI mouse and injected one hundred thousand of BD-MSC along the femoral artery to leg muscle. We validated stem cell survival, blood vessel formation, leg muscle condition and fibrosis compared by saline injected mice 28 days later. In this study, BD-MSC cultured medium treatment increased migration and tube formation of HUVEC and BDMSC injection had an effective blood vessel formation in FeCl3 treated CLI. As well as blood vessel formation, limb salvage rate also improved and fibrosis area statistically decreased in BD-MSC injected mice. In conclusion, bone marrow derived mesenchymal stem cell improved not only blood
김종순,변대석,김형락,김두상,신태선,김흥윤,오명주 한국수산과학회 2004 Fisheries and Aquatic Sciences Vol.7 No.2
Total phosphorus contents in rice bran and soybean meal were determined to be 5.81 and 2.77%, respectively, and 97.2% of phosphorus in rice bran and 66.4% in soybean meal were presented as phytate phosphorus. Optimum pH condition for hydrolysis of phytate in rice bran and soybean was determined to be in the pH range of 3.7 and 5.3. The highest activity of phytase for hydrolysis of phytate in both samples was determined to be at 55℃ for rice bran and 55-60℃ for soybean. Hydrolysis of phytate in soybean meal at pH 5.0 increased with the co-reaction or consecutive reaction with protease; however, in rice bran hydrolysis decreased with co-reaction with protease. Phytate degradation of soybean meal in the presence of pepsin at pH 2.5 showed higher than that of rice bran. Phytate degradation of rice bran in the presence of trypsin or pancreatin at pH 7.0 increased the activity around 2-times compared with the activity in the absence of trypsin or pancreatin. The results of this study suggest that hydrolysis of phytate in rice bran or soybean meal with phytase and protease may provide an alternative process for the preparation of aquacultural feed with a low level of organic phosphorus.
양혜원,손명주,Junwon Choi,오세연,전유진,변경희,유보미 한국수산과학회 2020 Fisheries and Aquatic Sciences Vol.23 No.3
Brown alga (Ishige okamurae; IO) dietary supplements have been reported to possess anti-diabetic properties. However, the effects of IO supplements have not been evaluated on glucose metabolism in the pancreas and skeletal muscle. C57BL/6 N male mice (age, 7 weeks) were arranged in five groups: a chow diet with 0.9% saline (NFD/saline group), high-fat diet (HFD) with 0.9% saline (HFD/saline group). high-fat diet with 25 mg/kg IO extract (HFD/25/IOE). high-fat diet with 50 mg/kg IO extract (HFD/50/IOE), and high-fat diet with 75 mg/kg IO extract (HFD/ 75/IOE). After 4 weeks, the plasma, pancreas, and skeletal muscle samples were collected for biochemical analyses. IOE significantly ameliorated glucose tolerance impairment and fasting and 2 h blood glucose level in HFD mice. IOE also stimulated the protein expressions of the glucose transporters (GLUTs) including GLUT2 and GLUT4 and those of their related transcription factors in the pancreases and skeletal muscles of HFD mice, enhanced glucose metabolism, and regulated blood glucose level. Our results suggest Ishige okamurae extract may reduce blood glucose levels by improving glucose metabolism in the pancreas and skeletal muscle in HFD-induced diabetes.