KR101742096B1 - Composition with chrysin for suppression of diabetic retinopathy - Google Patents
Composition with chrysin for suppression of diabetic retinopathy Download PDFInfo
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- KR101742096B1 KR101742096B1 KR1020160155692A KR20160155692A KR101742096B1 KR 101742096 B1 KR101742096 B1 KR 101742096B1 KR 1020160155692 A KR1020160155692 A KR 1020160155692A KR 20160155692 A KR20160155692 A KR 20160155692A KR 101742096 B1 KR101742096 B1 KR 101742096B1
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- KR
- South Korea
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- chrysin
- diabetic
- retinopathy
- endothelial cells
- hrmvec
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Abstract
본 발명은 크리신(chrysin)을 포함하는 당뇨병성 망막병증 억제용 조성물에 관한 것으로, 본 발명에 따른 크리신을 함유한 조성물을 이용함으로써, 망막 미세 혈관 내피세포에서 혈관 신생에 관여하는 단백질인 HIF-1α(Hypoxia-inducible factor 1-alpha)와, VEGF(Vascular endothelial growth factor)생성을 감소시키고, 고혈당에 노출시킨 망막 미세 혈관 내피세포의 세포사멸과 제2형 당뇨병 동물모델에서의 망막의 비정상적인 신생혈관(Acellular capillary) 형성 및 망막 혈관의 출혈을 억제시킴으로써, 당뇨합병증으로 초래되는 당뇨병성 망막병증을 개선시킬 수 있다. The present invention relates to a composition for inhibiting diabetic retinopathy comprising chrysin. By using a composition containing a chrysin according to the present invention, it is possible to provide a composition for inhibiting angiogenesis in a retinal microvascular endothelial cell, 1α (hypoxia-inducible factor 1-alpha) and VEGF (Vascular Endothelial Growth Factor) production, and the apoptosis of retinal microvascular endothelial cells exposed to hyperglycemia and the abnormal neovascularization of the retina in the type 2 diabetic animal model (Acellular capillary) formation and hemorrhage of the retinal vessels, diabetic retinopathy caused by diabetic complications can be improved.
Description
본 발명은 크리신을 함유하는 당뇨병성 망막병증 억제용 조성물 대한 것으로, 더욱 상세하게는 망막 미세 혈관 내피세포의 증식으로 인한 비정상적인 혈관신생(angiogenesis)으로 인한 당뇨병성 망막병증의 주요한 증상인 망막의 출혈 및 혈관신생 관련인자인, HIF-1α(Hypoxia-inducible factor 1-alpha)와 VEGF(Vascular endothelial growth factor) 생성을 감소시키고, 망막의 비정상적인 신생혈관(Acellular capillary) 형성을 억제하며, 고혈당 노출로 인한 망막 미세 혈관 내피세포의 사멸을 억제시킬 수 있는 크리신을 유효성분으로 함유하는 당뇨병성 망막병증 억제용 조성물에 대한 것이다.The present invention relates to a composition for inhibiting diabetic retinopathy comprising chrysin, and more particularly to a composition for inhibiting diabetic retinopathy, which is a major symptom of diabetic retinopathy caused by abnormal angiogenesis caused by proliferation of retinal microvascular endothelial cells, (HIF-1α) and Vascular Endothelial Growth Factor (VEGF), inhibits abnormal angiogenesis of the retina, and inhibits angiogenesis associated with hyperglycemic exposure The present invention relates to a composition for inhibiting diabetic retinopathy comprising as an active ingredient chrysin capable of inhibiting the death of microvascular endothelial cells.
당뇨병은 인슐린의 분비량이 부족하거나 정상적인 기능이 이루어지지 않는 등의 대사질환의 일종으로, 혈중 포도당의 농도가 높아지는 고혈당을 특징으로 하는데, 고혈당으로 인하여 여러 증상 및 징후를 일으키고 소변으로 포도당을 배출하게 된다. 또한, 당뇨병 환자가 장기간 고혈당에 노출되었을 경우에는 말초신경증, 자율신경증, 대 혈관 합병증, 미세 혈관 합병증을 비롯하여 기타 만성합병증을 초래할 수 있다. Diabetes mellitus is a type of metabolic disease that lacks insulin secretion or does not function normally. It is characterized by hyperglycemia in which the concentration of glucose in the blood increases. Hyperglycemia causes various symptoms and signs and discharges glucose into the urine . In addition, when diabetic patients are exposed to prolonged hyperglycemia, they may cause peripheral neuropathy, autonomic neuropathy, macrovascular complications, microvascular complications, and other chronic complications.
당뇨병성 망막병증은 이러한 당뇨병으로 인한 만성합병증의 하나로 망막의 미세 혈관이 손상되었을 때 나타나며, 전 세계적으로 중요한 실명 원인 중의 하나이다. 당뇨병성 망막병증은 크게 비증식성 망막병증과 증식성 망막병증으로 분류할 수 있으며, 비증식성 망막병증은 망막의 작은 혈관들이 약해져서 혈청이 잘 새거나 혈관이 막혀서 영양 공급이 중단되는 상태를 말한다. 당뇨병성 망막병증은 서서히 발생되고, 시력감퇴가 점진적으로 일어난다는 특징을 가지고 있는데, 이는 당뇨망막병증의 초기 소견이라고 할 수 있다. Diabetic retinopathy is one of the chronic complications of diabetes, which occurs when the microvessels of the retina are damaged and is one of the causes of blindness globally. Diabetic retinopathy can be classified into non-proliferative retinopathy and proliferative retinopathy. Non-proliferative retinopathy refers to a state in which small blood vessels in the retina become weak and serum is well leaked or blood vessels are blocked and nutrition is stopped. Diabetic retinopathy is characterized by gradual development of vision and gradual deterioration of vision, which is an early finding of diabetic retinopathy.
증식성 망막병증은 고혈당으로 인한 혈액순환 기능의 저하로 인하여, 망막에 비정상적인 신생혈관이 생기는 것을 지칭하는데, 적절한 치료를 받지 않을 경우, 신생 혈관으로부터 발생하는 출혈에 의해 5년 이내에 실명하게 되는 무서운 합병증으로써, 당뇨 망막병증의 후기 소견이라고 할 수 있다.Proliferative retinopathy refers to the occurrence of abnormal neovascularization in the retina due to a decrease in blood circulation due to hyperglycemia. If the patient does not receive appropriate treatment, the patient will suffer from a severe complication , Suggesting a late finding of diabetic retinopathy.
한편, 크리신(chrysin)은 식물에 광범위하게 분포되어 있는 플라보노이드의 일종으로, 항산화, 항염증, 항암, 항알레르기 작용 등 다양한 생리활성이 보고되어 있다. 현재는 근육강화제로 시판되어 널리 쓰이고 있는 성분 중의 하나이며, 최근에는 항불안 효과, 항경련 효능에 대한 연구가 알려져 있으며, 크리신 유도체들을 이용한 항 당뇨 활성에 대한 많은 연구가 진행 중에 있다.On the other hand, chrysin is a type of flavonoid widely distributed in plants, and various physiological activities such as antioxidation, anti-inflammation, anti-cancer and antiallergic action have been reported. Recently, studies on anti-anxiety effect and anti-seizure efficacy have been known, and many studies on anti-diabetic activity using chrysin derivatives are under way.
현재까지 당뇨 합병증의 하나인 당뇨병성 망막병증의 예방 및 치료를 위한 연구가 활발하게 이루어지고 있으며, 장기간 고혈당 노출되어 발생하는 당뇨병성 망막병증의 완화를 위한 다양한 연구가 진행 중에 있다. 그러나, 식이 성분을 포함한 천연 성분을 이용하여 망막병증을 예방하고 치료하는 연구는 미흡한 실정이다. Various studies for the prevention and treatment of diabetic retinopathy, which is one of the complications of diabetes, have been actively conducted, and various studies are under way to alleviate diabetic retinopathy caused by prolonged exposure to hyperglycemia. However, studies on the prevention and treatment of retinopathy using natural ingredients including dietary ingredients are insufficient.
따라서, 망막 미세 혈관 내피세포의 증식으로 인한 비정상적인 혈관신생(angiogenesis)과 이로 인한 망막의 출혈 및 혈관신생 관련인자들의 생성을 감소시키고, 망막의 비정상적인 신생혈관(Acellular capillary) 형성을 억제하며, 고혈당 노출로 인한 망막 미세 혈관 내피세포의 사멸을 억제시킬수 있는 천연성분에 대한 연구가 시급한 실정이다.Therefore, it is possible to reduce abnormal angiogenesis due to proliferation of retinal microvascular endothelial cells, thereby reducing the production of retinal hemorrhage and angiogenesis-related factors, inhibiting the formation of acellular capillary cells in the retina, , Which has been known to inhibit retinal microvascular endothelial cell death, is urgently needed.
본 발명은 만성 고혈당으로 인한 당뇨병성 망막병증을 개선시킬 수 있는 새로운 소재를 개발하여 제공하고자 한다. The present invention provides a novel material capable of improving diabetic retinopathy due to chronic hyperglycemia.
본 발명은 크리신(chrysin)을 포함하는 당뇨합병성 망막병증 개선용 식품 조성물을 제공한다. The present invention provides a food composition for improving diabetic complicated retinopathy comprising chrysin.
본 발명의 당뇨합병성 망막병증 개선용 식품 조성물에 있어서, 상기 크리신은, 바람직하게 만성 고혈당에 노출된 망막 미세 혈관 내피세포(HRMVEC)에서 비정상적인 혈관 신생에 관여하는 단백질인 HIF-1α(Hypoxia-inducible factor 1-alpha)와 VEGF(Vascular endothelial growth factor) 생성을 감소시키는 것일 수 있다. In the food composition for improving diabetic complication retinopathy of the present invention, the above-mentioned chrysin is preferably selected from the group consisting of Hypoxia-inducible (HIF-1α), a protein involved in abnormal angiogenesis in retinal microvascular endothelial cells (HRMVEC) factor 1-alpha) and VEGF (vascular endothelial growth factor) production.
본 발명의 당뇨합병성 망막병증 개선용 식품 조성물에 있어서, 상기 크리신은, 바람직하게 제2형 당뇨병에서 망막의 비정상적인 신생혈관(Acellular capillary) 형성을 억제하는 것일 수 있다. In the food composition for improving diabetic complication retinopathy of the present invention, the chrysin may preferably inhibit the formation of abnormal acellular capillaries of the retina in
본 발명의 당뇨합병성 망막병증 개선용 식품 조성물에 있어서, 상기 크리신은, 바람직하게 만성 고혈당에 노출된 망막 미세 혈관 내피세포(HRMVEC)의 세포사멸을 억제하는 것일 수 있다. In the food composition for improving diabetic complication retinopathy of the present invention, the above-mentioned chrysin may preferably inhibit apoptosis of retinal microvascular endothelial cells (HRMVEC) exposed to chronic hyperglycemia.
본 발명은 크리신(chrysin)을 포함하는 당뇨합병성 망막병증 치료 또는 예방용 약학 조성물을 제공한다.The present invention provides a pharmaceutical composition for treating or preventing diabetic complicated retinopathy comprising chrysin.
본 발명의 당뇨합병성 망막병증 개선용 식품 조성물에 있어서, 상기 크리신은, 바람직하게 만성 고혈당에 노출된 망막 미세 혈관 내피세포(HRMVEC)에서 비정상적인 혈관 신생에 관여하는 단백질인 HIF-1α(Hypoxia-inducible factor 1-alpha)와 VEGF(Vascular endothelial growth factor) 생성을 감소시키는 것일 수 있다. In the food composition for improving diabetic complication retinopathy of the present invention, the above-mentioned chrysin is preferably selected from the group consisting of Hypoxia-inducible (HIF-1α), a protein involved in abnormal angiogenesis in retinal microvascular endothelial cells (HRMVEC) factor 1-alpha) and VEGF (vascular endothelial growth factor) production.
본 발명의 당뇨합병성 망막병증 개선용 식품 조성물에 있어서, 상기 크리신은, 바람직하게 제2형 당뇨병에서 망막의 비정상적인 신생혈관(Acellular capillary) 형성을 억제하는 것일 수 있다. In the food composition for improving diabetic complication retinopathy of the present invention, the chrysin may preferably inhibit the formation of abnormal acellular capillaries of the retina in
본 발명의 당뇨합병성 망막병증 개선용 식품 조성물에 있어서, 상기 크리신은, 바람직하게 만성 고혈당에 노출된 망막 미세 혈관 내피세포(HRMVEC)의 세포사멸을 억제하는 것일 수 있다. In the food composition for improving diabetic complication retinopathy of the present invention, the above-mentioned chrysin may preferably inhibit apoptosis of retinal microvascular endothelial cells (HRMVEC) exposed to chronic hyperglycemia.
본 발명에 따른 크리신을 함유한 조성물을 이용함으로써, 망막 미세 혈관 내피세포에서 혈관 신생에 관여하는 단백질인 HIF-1α(Hypoxia-inducible factor 1-alpha)와 VEGF(Vascular endothelial growth factor) 생성을 감소시키고, 고혈당에 노출시킨 망막 미세 혈관 내피세포의 세포사멸과 제2형 당뇨병 동물모델에서의 망막의 비정상적인 신생혈관(Acellular capillary) 형성 및 망막 혈관의 출혈을 억제시킴으로써 당뇨합병증으로 초래되는 당뇨병성 망막병증을 개선시킬 수 있다. By using the composition containing the chrysin according to the present invention, it is possible to reduce the production of hypoxia-inducible factor 1-alpha (HIF-1α) and vascular endothelial growth factor (VEGF), proteins involved in angiogenesis in retinal microvascular endothelial cells , Diabetic retinopathy caused by diabetic complications by inhibiting retinal microvascular endothelial cell apoptosis induced by hyperglycemia, abnormal acellular capillary formation of the retina in the
도 1은 크리신을 망막 미세 혈관 내피세포에 처리하여 배양하였을 때, 만성 고혈당에서 혈관 신생에 관여하는 단백질인 HIF-1α(Hypoxia-inducible factor 1-alpha)와, VEGF(Vascular endothelial growth factor)의 발현 및 분비 억제 활성을 나타낸 웨스턴 블롯 분석결과이며, 이것을 덴시토미터(densitometer)로 측정하여 정량화한 것이다.
도 2는 크리신을 제2형 당뇨병 동물모델에 10주간 10mg/kg의 농도로 처리한 후 동물 망막조직에서 나타나는 비정상적인 혈관(Acellular capillary)의 생성을 억제하는 활성을 나타낸 헤마톡실린 에오신 염색 분석 결과이며, 이것을 정량화 하여 그래프로 나타낸 것이다.
도 3은 크리신을 망막 미세 혈관 내피세포에 처리하여 배양하였을 때, 만성 고혈당 상태에서 나타나는 세포사멸 억제 효과를 나타내는 TUNEL assay 결과이며, 이것을 덴시토미터로 측정하여 정량화한 것이다. Figure 1 shows the expression of Hypoxia-inducible factor 1-alpha (HIF-1α), a protein involved in angiogenesis in chronic hyperglycemia, and the expression of VEGF (Vascular Endothelial Growth Factor), when cultured by treating crecin with retinal microvascular endothelial cells And secretion inhibitory activity, which was quantified by measuring with a densitometer.
FIG. 2 is a result of hematoxylin eosin staining analysis showing the activity of inhibiting the formation of abnormal blood vessels (Acellular capillary) in animal retina tissue after treatment with 10 mg / kg of chrysin in a
FIG. 3 shows the results of TUNEL assay showing the effect of inhibiting apoptosis in chronic hyperglycemic state when cultured by treating crecin with retinal microvascular endothelial cells, which is quantified by measuring with a densitometer.
본 발명은 당뇨합병증으로 인해 초래되는 당뇨병성 망막병증 억제 활성을 갖는 크리신을 유효성분으로 함유하는 조성물에 대한 것이다. 더욱 상세하게는 당뇨합병증으로 초래되는 당뇨병성 망막병증에 직접적으로 관련된 인간 망막 미세 혈관 내피세포 (Human Retinal Microvascular Endothelial cell, HRMVEC)를 고혈당에 노출시키고, 여기에 크리신을 처리하여 배양함으로써, 비정상적인 혈관 신생에 관여하는 단백질인 HIF-1α(Hypoxia-inducible factor 1-alpha)와, VEGF(Vascular endothelial growth factor)생성을 감소시키고, 제2형 당뇨병 동물모델을 이용하여 10주간 10mg/kg의 chrysin을 경구투여한 후 망막의 비정상적인 신생혈관(Acellular capillary) 형성의 억제를 확인하였으며, 고혈당에 노출시킨 망막 미세 혈관 내피세포의 세포사멸과 이로 인한 동물에서의 망막 혈관의 출혈을 억제시킴으로써, 만성 고혈당의 결과로 나타나는 당뇨병성 망막병증을 개선시킬 수 있는 크리신을 유효성분으로 함유하는 조성물에 대한 것이다. The present invention relates to a composition containing as an active ingredient chrysin having an activity to inhibit diabetic retinopathy caused by diabetic complications. More specifically, human retinal microvascular endothelial cell (HRMVEC), which is directly related to diabetic retinopathy caused by diabetic complications, is exposed to hyperglycemia and cultured by treatment with chrysin, whereby abnormal angiogenesis (HGF-1α) and VEGF (vascular endothelial growth factor) production, and 10 mg / kg chrysin was orally administered for 10 weeks using an animal model of
크리신은 하기 화학식 1과 같은 구조를 가지고 있고, 통상적인 방법에 따라 제조하여 사용하거나, 시판되는 것을 구입하여 사용할 수 있다.The chrysene has a structure as shown in the following Chemical Formula 1, and can be prepared by using a conventional method or commercially available.
[화학식 I](I)
크리신(5,7-Dihydroxy-2-phenyl-4H-chromen-4-one)(5,7-Dihydroxy-2-phenyl-4H-chromen-4-one)
본 발명에 의한 당뇨합병증으로 인해 초래되는 망막병증에 억제 활성을 갖는 크리신(chrysin)을 유효성분으로 함유한 조성물은 그 제형에 있어서 특별히 한정되지 않지만, 바람직하게는 식품조성물 또는 약학조성물일 수 있다.The composition containing chrysin having an inhibitory activity on retinopathy caused by diabetic complications according to the present invention as an active ingredient is not particularly limited in its formulation but may be preferably a food composition or a pharmaceutical composition .
상기 약학조성물은 유효성분 이외에 약제학적으로 허용 가능한 담체, 희석제 또는 부형제를 더욱 포함할 수 있다. 사용 가능한 담체, 부형제 또는 희석제로는, 락토즈, 덱스트로즈, 수크로즈, 솔비톨, 만니톨, 자이리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘포스페이트, 칼슘실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로즈, 폴리비닐피롤리돈, 물, 메틸하이드록시벤조에이트, 프로필하이드록시벤조에이트, 탈크, 마그네슘스테아레이트 및 광물유가 있으며, 이중 선택되는 하나 이상을 사용할 수 있다. 또한, 상기 약학조성물이 약제인 경우, 충진제, 항응집제, 윤활제, 습윤제, 향료, 유화제 또는 방부제 중 선택되는 하나 이상을 추가적으로 포함할 수 있다.The pharmaceutical composition may further comprise, in addition to the active ingredient, a pharmaceutically acceptable carrier, diluent or excipient. Examples of usable carriers, excipients or diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and one or more selected from among them may be used. When the pharmaceutical composition is a pharmaceutical, it may further include at least one selected from a filler, an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent or an antiseptic agent.
본 발명 약학조성물의 제형은 사용방법에 따라 바람직한 형태일 수 있으며, 특히 포유동물에 투여된 후 활성 성분의 신속, 지속 또는 지연된 방출을 제공할 수 있도록 당업계에 공지된 방법을 채택하여 제형화 하는 것이 좋다. 구체적인 제형의 예로는 경고제(PLASTERS), 과립제(GRANULES), 로션제(LOTIONS), 리니멘트제(LINIMENTS), 리모나데제(LEMONADES), 방향수제(AROMATIC WATERS), 산제(POWDERS), 시럽제(SYRUPS), 안연고제(OPHTHALMIC OINTMENTS), 액제(LIQUIDS AND SOLUTIONS), 에어로솔제(AEROSOLS), 엑스제(EXTRACTS), 엘릭실제(ELIXIRS), 연고제(OINTMENTS), 유동엑스제(FLUIDEXTRACTS), 유제(EMULSIONS), 현탁제(SUSPENSIONS), 전제(DECOCTIONS), 침제(INFUSIONS), 점안제(OPHTHALMIC SOLUTIONS), 정제(TABLETS), 좌제(SUPPOSITORIES), 주사제(INJECTIONS), 주정제(SPIRITS), 카타플라스마제(CATAPLSMA), 캅셀제(CAPSULES), 크림제(CREAMS), 트로키제(TROCHES), 틴크제(TINCTURES), 파스타제(PASTES), 환제(PILLS), 연질 또는 경질 젤라틴 캅셀 중 선택되는 어느 하나일 수 있다. The formulations of the pharmaceutical compositions of the present invention may be in their preferred form depending on the method of use, and may be formulated employing methods known in the art to provide rapid, sustained or delayed release of the active ingredient, It is good. Examples of specific formulations include, but are not limited to, PLASTERS, GRANULES, LOTIONS, LINIMENTS, LEMONADES, AROMATIC WATERS, POWDERS, SYRUPS, OPHTHALMIC OINTMENTS, LIQUIDS AND SOLUTIONS, AEROSOLS, EXTRACTS, ELIXIRS, OINTMENTS, FLUIDEXTRACTS, EMULSIONS, ), Suspensions, DECOCTIONS, INFUSIONS, OPHTHALMIC SOLUTIONS, TABLETS, SUPPOSITORIES, INJECTIONS, SPIRITS, CATAPLSMA, ), Capsules, CREAMS, TROCHES, TINCTURES, PASTES, PILLS, soft or hard gelatin capsules.
본 발명 약학조성물의 투여량은 투여방법, 복용자의 연령, 성별, 체중, 및 질환의 중증도 등을 고려하여 결정하는 것이 좋다. 일 예로, 본 발명의 상기 약학조성물은 유효성분인 크리신을 기준으로 하였을 때 1일 0.00001 내지 100 ㎎/㎏(체중)으로 1회 이상 투여가능하다. 그러나 상기의 투여량은 예시하기 위한 일 예에 불과하며, 복용자의 상태에 따라 의사의 처방에 의해 변화될 수 있다.The dosage of the pharmaceutical composition of the present invention is preferably determined in consideration of the administration method, the age, sex, weight, and severity of the disease, etc. of the recipient. For example, the pharmaceutical composition of the present invention can be administered at least once at a dose of 0.00001 to 100 mg / kg (body weight) per day based on the active ingredient chrysin. However, the dosage is only an example and may be changed by a doctor's prescription depending on the condition of the recipient.
한편, 본 발명의 식품조성물은 일 예로, 육류, 곡류, 카페인 음료, 일반음료, 초콜렛, 빵류, 스낵류, 과자류, 사탕, 피자, 젤리, 면류, 껌류, 유제품류, 아이스크림류, 알코올성 음료, 술, 비타민 복합제 및 그 밖의 건강보조식품류 중 선택되는 어느 하나일 수 있으나, 이에 한정되는 것은 아니다. 본 발명의 식품 조성물에 첨가되는 크리신의 양은 식품의 제형을 해치지 않는 범주 및 독성을 불러일으키지 않는 범주 내에서 그 양을 다양하게 하여 첨가될 수 있다. Meanwhile, the food composition of the present invention can be used as a food composition, such as meat, cereal, caffeinated beverage, general beverage, chocolate, bread, snack, confectionery, candy, pizza, jelly, noodle, dairy, ice cream, Vitamin complex, and other health supplement foods. However, the present invention is not limited thereto. The amount of chrysin added to the food composition of the present invention may be added in various amounts within the category that does not adversely affect the formulation of the food and does not cause toxicity.
이하, 본 발명의 내용을 하기 실시예를 통해 더욱 구체적으로 설명하고자 한다. 다만, 본 발명의 권리범위가 하기 실시예에만 한정되는 것은 아니고, 그와 등가의 기술적 사상의 변형까지를 포함한다.Hereinafter, the present invention will be described more specifically with reference to the following examples. However, the scope of the present invention is not limited to the following embodiments, and includes modifications of equivalent technical ideas.
[[ 비교예Comparative Example 1 : 세포배양 실험 1 (인간 망막 미세 혈관 내피세포)] 1: Cell Culture Experiment 1 (Human Retinal Microvascular Endothelial Cells)]
인간 망막 미세 혈관 내피세포(HRMVEC)는 Cell System (Kirkland, WA)에서 구입하여 배양하였다. HRMVEC는 10% 소태아혈청(Fetal Bovine Serum, FBS)과 2mM 글루타민(Glutamine), 100 U/ml 페니실린(Penicillin), 100 mg/ml 스트렙토마이신(Streptomycin)이 첨가되어 있는 Media199(M199, Sigma Co., St. Louis, MO, USA) 배지에, 0.75 μg/ml EGF(human epidermal growth factor) 및 75 μg/ml 하이드로코르티손(hydrocortisone)을 첨가하여 사용하였으며 37℃, 5% 이산화탄소(CO2) 조건에서 배양하였다. Human retinal microvascular endothelial cells (HRMVEC) were purchased from the Cell System (Kirkland, WA) and cultured. HRMVEC was prepared from Media199 (M199, Sigma Co.) supplemented with 10% Fetal Bovine Serum (FBS), 2 mM glutamine, 100 U / ml penicillin and 100 mg / ml streptomycin. , Human epidermal growth factor (EGF) and 75 μg / ml hydrocortisone were added to the medium at 37 ° C in 5% CO 2 Lt; / RTI >
[[ 실시예Example 1 : 세포배양 실험 2 (고혈당으로 인한 당뇨병성 망막병증)] 1: Cell culture experiment 2 (diabetic retinopathy due to hyperglycemia)]
만성 고혈당으로 인한 당뇨병성 망막병증의 증상을 유도하기 위하여 인간 망막 미세 혈관 내피세포(HRMVEC)를 33mM 글루코오스(Glucose)를 첨가한 배지에서 배양하고 크리신(chrysin)을 농도별(1~20μM)로 처리하여 함께 배양하였다. 또한, 삼투 대조군으로 5.5mM 글루코오스(Glucose)와 27.5mM 마니톨(Mannitol)이 함유된 배지에서 망막 미세 혈관 내피세포를 2일간 배양하였다.Human retinal microvascular endothelial cells (HRMVEC) were cultured in a medium supplemented with 33 mM glucose to induce symptoms of diabetic retinopathy due to chronic hyperglycemia, and chrysin was added at a concentration of 1 to 20 μM And cultured together. In addition, retinas microvascular endothelial cells were cultured for 2 days in a medium containing 5.5 mM glucose and 27.5 mM mannitol as an osmotic control group.
[[ 실시예Example 2 : 혈관신생 2: angiogenesis 관련인자(angiogenic factor)인An angiogenic factor HIF-1α( HIF-1? ( HypoxiaHypoxia -inducible factor 1-alpha)와 -inducible factor 1-alpha) and VEGFVEGF (Vascular endothelial growth factor) 발현 억제] (Vascular Endothelial Growth Factor)
인간 망막 미세 혈관 내피세포(HRMVEC)의 혈관신생 관련인자(angiogenic factor)인 HIF-1α와, VEGF 발현 정도를 측정하기 위해 상기 비교예 1, 실시예 1에서 배양처리된 망막 미세 혈관 내피세포(HRMVEC)를, 1% β-Mercaproethanol, 1M β-Glycerophosphate, 0.1M Na3VO4, 0.5M 불화소다(NaF)와 프로테아제 인히비터 칵테일 (Protease Inhibitor Cocktail)을 포함한 완충액(Buffer)으로 분해(Lysis)시켜, 3000rpm에서 10분간 원심분리하여 얻은 상층액을 이용하였다. HIF-1α, an angiogenic factor of human retinal microvascular endothelial cells (HRMVEC), and retinal microvascular endothelial cells (HRMVECs) cultured in Comparative Example 1 and Example 1 were measured to measure the degree of VEGF expression ) Was lysed with a buffer containing 1% β-mercaproethanol, 1 M β-Glycerophosphate, 0.1 M Na 3 VO 4 , 0.5 M sodium fluoride (NaF) and a Protease Inhibitor Cocktail , And centrifuged at 3000 rpm for 10 minutes to obtain a supernatant.
이렇게 얻은 상층액을 10% 또는 8% SDS-PAGE 겔(Gel) 상에서 70V로 전기영동 하였다. 겔(Gel) 상에 있는 단백질은 니트로셀룰로우스 막(Nitrocellulose Membrane)으로 전이시킨 다음, 비특이적인 결합을 방지하기 위하여 3% 소혈청 알부민 (Bovine Serum Albumin, BSA)에 3시간 동안 블록킹(blocking) 하였다. The supernatant thus obtained was electrophoresed at 70 V on 10% or 8% SDS-PAGE gel. The protein on the gel was transferred to a nitrocellulose membrane and blocked with 3% bovine serum albumin (BSA) for 3 hours to prevent nonspecific binding. Respectively.
HIF-1α의 일차항체(Monoclonal Anti-mouse Antibody, Abcam Biochemicals, Cambridge, UK)와, VEGF의 일차항체(Polyclonal Anti-Goat Antibody, R&D systems, Minneapolis, MN)로 니트로셀룰로우스 막(Nitrocellulose membrane)과 교반하면서 3시간 배양한 후, 4℃에서 하룻밤 배양하고 다시 실온에서 3시간 배양한 후 이차항체인 Anti-mouse 와 Anti-goat Horseradish Peroxidase (Jackson ImmunoResearch Lab., West Grove, PA, USA)로 1시간 동안 배양하였다. Nitrocellulose membranes were prepared with HIF-1α primary antibody (Monoclonal Anti-Mouse Antibody, Abcam Biochemicals, Cambridge, UK) and VEGF primary antibody (Polyclonal Anti-Goat Antibody, R & D systems, Minneapolis, MN) And anti-goat Horseradish peroxidase (Jackson ImmunoResearch Lab., West Grove, PA, USA) were incubated for 3 hours at room temperature. Lt; / RTI >
니트로셀룰로스 막(Nitrocellulose Membrane)에 전이된 단백질은 'Immobilon western chemiluminescent horseradish peroxidase substrate (Millipore Corp., Billerica, MA, USA)'로 검출하여 비교예 1에서 배양 처리된 정상 망막 미세 혈관 내피세포(HRMVEC)와 실시예 1에서 크리신(chrysin)을 1μM, 10μM, 20μM 로 각각 처리한 세포 샘플을 X-ray 필름으로 촬영하였다.The protein transferred to the nitrocellulose membrane was detected by Immobilon western chemiluminescent horseradish peroxidase substrate (Millipore Corp., Billerica, MA, USA), and the normal retinal microvascular endothelial cells (HRMVEC) And chrysin at 1 μM, 10 μM, and 20 μM, respectively, in Example 1 were photographed with X-ray film.
위 결과는 도 1에 나타내었다. 도 1에 도시된 바와 같이, 실시예 1의 삼투 대조군인 망막 미세 혈관 내피세포(5.5mM glucose, 27.5mM mannitol 대조군)는 HIF-1α와 VEGF 단백질의 발현 및 분비가 적었으나, 2일간 고혈당에 노출된 망막 미세 혈관 내피세포(HRMVEC)는 HIF-1α와 VEGF 단백질의 발현 및 분비가 현저하게 증가하였다. The above results are shown in Fig. As shown in FIG. 1, the osmotic control group of Example 1, retinal microvascular endothelial cells (5.5 mM glucose, 27.5 mM mannitol control) showed less HIF-1α and VEGF protein expression and secretion, but exposed to high glucose for 2 days The expression and secretion of HIF-1α and VEGF protein were significantly increased in the retinal microvascular endothelial cells (HRMVEC).
여기서, 2일간 고혈당에 노출된 HRMVEC에 크리신을 1μM 처리하였을 때 HIF-1α와 VEGF 단백질의 발현 및 분비에는 거의 변화가 없었으며, 크리신(chrysin)을 10μM 이상 처리하였을 때는 HIF-1α와 VEGF 단백질의 발현 및 분비가 점차 억제되는 것으로 나타났다.When HRMVEC exposed to hyperglycemia for 2 days was treated with 1 μM of chrysin, there was almost no change in the expression and secretion of HIF-1α and VEGF protein. When treated with chrysin at a concentration of 10 μM or more, HIF-1α and VEGF protein The expression and secretion of the protein were gradually suppressed.
따라서, 만성 고혈당에 노출된 HRMVEC에 크리신(chrysin)을 10μM 이상으로 처리하였을 경우에 혈관신생 관련인자인 HIF-1α(Hypoxia-inducible factor 1-alpha)와, VEGF(Vascular endothelial growth factor)생성을 억제시키는 효과가 뛰어난 것을 알 수 있었다. Thus, when HRMVEC exposed to chronic hyperglycemia was treated with chrysin at a concentration of 10 μM or more, the production of angiogenic factors such as Hip-1α (Hypoxia-inducible factor 1-alpha) and VEGF (vascular endothelial growth factor) It was found that the effect of suppressing the inhibition was excellent.
[[ 실시예Example 3 : 비정상적인 혈관(Acellular capillary) 생성 억제] 3: Inhibit the formation of abnormal blood vessels (Acellular capillary)
제2형 당뇨병 쥐 모델인 db/db mice를 이용하여 실제로 크리신이 당뇨병성 망막병증을 억제하는 역할을 수행하는지 알아보기 위해 동물실험을 실시하였다.Animal experiments were performed to investigate whether clincin effectively inhibits diabetic retinopathy using db / db mice, a
db/db 마우스는 렙틴수용체에 돌연변이가 생겨 지방세포에서 분비하는 호르몬인 렙틴의 신호전달이 일어나지 않아 비만과 제2형 당뇨병이 발생하는 동물모델로서 제2형 당뇨병 연구에 많이 사용되는 동물모델이다. db / db mouse is an animal model of obesity and
동물을 3가지 그룹으로 나누어 대조군(control) 그룹인 db/m mice, 제2형 당뇨그룹인 db/db mice, 제2형 당뇨모델에 크리신을 투여한 db/db-chrysin mice 실험을 실시하였다.Animals were divided into three groups and tested in control / db / m mice, db / db mice in
동물실험은 한림대학교 실험동물실에서 이루어졌으며, 낮과 밤이 12시간의 간격으로 조절되었으며, 생육온도가 23±1℃, 습도가 50±10% 그리고 병해충의 위협이 없는 조건에서 사육하였다. Animal experiments were carried out in the laboratory animal room of Hallym University. The day and night were controlled at intervals of 12 hours, and the growth temperature was 23 ± 1 ℃, the humidity was 50 ± 10%
동물은 10주간 사육되었으며, 크리신 처리 그룹은 10mg/kg의 농도로 10주간 경구투여한 후, 눈을 적출하여 4% 파라포름알데하이드에 24시간 동안 고정시킨 후 증류수에 담가 24시간 배양하였다. 그 후 망막을 분리하여, 3% 트립신에 0.1 M Tris 버퍼 (pH 7.8)가 함유된 시약에서 1시간 동안 배양하여 혈관만 분리해낸 후 슬라이드 글라스 위에서 건조시켰다. 건조된 혈관은 헤마톡실린&에오신 염색(H&E staining)을 실시한 후 현미경으로 관찰하였다. The animals were fed for 10 weeks, and the chrysin treatment group was orally administered at a concentration of 10 mg / kg for 10 weeks. The eyes were then removed and fixed in 4% paraformaldehyde for 24 hours and then immersed in distilled water for 24 hours. Then, the retina was separated and cultured in a reagent containing 0.1 M Tris buffer (pH 7.8) in 3% trypsin for 1 hour to separate the blood vessels and then dried on a slide glass. The dried blood vessels were subjected to hematoxylin & eosin staining (H & E staining) and then observed with a microscope.
위 결과는 도 2에 나타내었다. 도 2에 도시된 바와 같이, 실시예 3의 대조군 db/m control 그룹은 망막 혈관이 정상적인 패턴을 보이고 있으나, 제2형 당뇨병 그룹인 db/db mice 그룹에서는 화살표 표시된 부분과 같이 비정상적인 혈관(Acellular capillary)들이 생성되어 있는 것을 확인할 수 있었다. 반면에 10mg/kg으로 10주간 크리신을 투여한 동물 모델에서는 대조군 그룹과 유사한 정상적인 혈관의 패턴을 보였으며, 당뇨모델에서 확인되는 비정상적인 혈관들이 보이지 않는 것을 확인하였다. The above results are shown in Fig. As shown in FIG. 2, the control db / m control group of Example 3 shows a normal pattern of retinal blood vessels. However, in the db / db mice group of the
따라서, 만성 고혈당에 노출된 제2형 당뇨병 동물 모델에 10mg/kg의 농도로 10주간 크리신을 투여한 경우에 동물 망막조직에서 생성되는 비정상적인 혈관(Acellular capillary)의 생성을 유의적으로 억제하는 것으로 나타나, 크리신은 당뇨병성 망막병증을 억제하는 데에 뛰어난 효과를 발휘하는 것을 알 수 있다.Thus, it was shown that when 10 mg / kg of chrysin was administered to a model of
[[ 실시예Example 4 : 고혈당 자극으로 인한 인간 망막 미세 혈관 내피세포( 4: Human retinal microvascular endothelial cells due to hyperglycemic stimulation HRMVECHRMVEC )의 세포사멸 억제 효과 확인]) ≪ / RTI >
고혈당 자극으로 인한 망막 미세 혈관 내피세포(HRMVEC)에서의 세포사멸 억제효과를 알아보기 세포사멸을 확인하는 대표적인 방법인 TUNEL assay를 실시하였다. 정상적인 세포와는 달리 고혈당의 자극을 받아 손상이 일어난 망막 미세 혈관 내피세포는 핵의 정상적인 DNA의 이중나선구조가 절단되어 DNA 조각들이 만들어지게 되는데, TUNEL assay는 세포사멸이 일어난 세포에서 절단된 DNA의 유리 3-OH말단에 terminal deoxynucleotidyl transferase(TdT)라는 효소를 이용하여 형광물질이 결합된 uridine triphosphate(UTP)를 특이적으로 표식한 후 현미경으로 확인하는 방법이다. Detection of cytotoxic effect on retinal microvascular endothelial cells (HRMVEC) due to hyperglycemic stimulation TUNEL assay, a representative method for confirming apoptosis, was performed. In contrast to normal cells, retinal microvascular endothelial cells, which are damaged by hyperglycemia, are cleaved by the normal double-stranded DNA structure of DNA, resulting in DNA fragments. Uridine triphosphate (UTP) conjugated with a fluorescent substance is specifically labeled with an enzyme called terminal deoxynucleotidyl transferase (TdT) at the 3-OH terminal of the free glass and then confirmed by a microscope.
비교예 1에서 배양 처리된 정상 인간 망막 미세 혈관 내피세포(HRMVEC)와 실시예 1에서 크리신(chrysin)을 1μM, 10μM, 20μM로 각각 처리한 세포 샘플을 형광현미경으로 촬영하였다.Cell samples obtained by treating normal human retinal microvascular endothelial cells (HRMVEC) cultured in Comparative Example 1 and 1 μM, 10 μM and 20 μM of chrysin in Example 1 were photographed by fluorescence microscope.
위 결과는 도 3에 나타내었다. 도 3에 도시된 바와 같이, 실시예 1의 삼투 대조군인 인간 망막 미세 혈관 내피세포(5.5mM glucose, 27.5mM mannitol 대조군)는 TUNEL assay 결과 핵에서 나타나는 DNA 조각의 염색 정도가 적었으나, 2일간 고혈당에 노출된 인간 망막 미세 혈관 내피세포(HRMVEC)에서는 핵에서 나타나는 DNA 조각들의 염색이 현저하게 증가하였다. 여기에 2일간 고혈당에 노출된 HRMVEC에 크리신(chrysin)을 1μM에서 20μM 처리하였을 때, DNA 조각이 염색되는 비율이 농도 의존적으로 점차 억제되는 것으로 나타났다. The above results are shown in Fig. As shown in FIG. 3, the degree of staining of DNA fragments in the nucleus of the human retinal microvascular endothelial cells (5.5 mM glucose, 27.5 mM mannitol control group) of Example 1 was low in the TUNEL assay. However, In human retinal microvascular endothelial cells (HRMVEC), the staining of DNA fragments in the nucleus was significantly increased. When HRMVEC exposed to high glucose for 2 days was treated with chrysin at 1 μM at 20 μM, the proportion of staining of DNA fragments was gradually inhibited in a concentration-dependent manner.
따라서, 만성 고혈당에 노출된 HRMVEC에 크리신(chrysin)을 10μM 이상으로 처리하였을 경우에 인간 망막 미세 혈관 내피세포에서 세포사멸을 억제하는데에 뛰어난 효과를 발휘하는 것을 알 수 있다.Therefore, it can be seen that when HRMVEC exposed to chronic hyperglycemia is treated with chrysin at 10 μM or more, it exerts an excellent effect in inhibiting apoptosis in human retinal microvascular endothelial cells.
[[
실시예Example
5 : 제2형 당뇨병 동물모델에서의 망막 혈관 출혈 억제 효과] 5: Inhibition of retinal vascular bleeding in an animal model of
제2형 당뇨병 쥐 모델인 db/db mice를 이용하여 10주간 10mg/kg의 농도로 10주간 경구투여한 후, 동물을 마취하여 20 mg/ml농도의 FITC-dextran을 마우스의 좌심실에 주사한 뒤 눈을 적출하여 4% 파라포름알데하이드에 10분동 동안 고정시킨 후 망막을 분리하여 슬라이드 글라스에 flat-mount 한 후 형광 현미경으로 관찰하였다.Db / db mice, a
위 결과는 도 4에 나타내었다. 도 4에 도시된 바와 같이 실시예 3의 대조군 db/m control 그룹은 망막 혈관에서의 초록색 염료인 FITC-dextran의 유출이 보이지 않는 정상적인 패턴을 보이고 있으나, 제2형 당뇨병 그룹인 db/db mice 그룹에서는 혈관 사이의 틈으로 화살표 표시된 부분과 같이 FITC-dextran의 유출 나타나는 것을 확인할 수 있었다. 반면에 10mg/kg으로 10주간 크리신을 투여한 동물 모델에서는 대조군 그룹과 유사하게 FITC-dextran의 유출이 보이지 않는 정상적인 혈관의 패턴을 보이는 것을 확인하였다. The above results are shown in Fig. As shown in FIG. 4, the control group db / m control group of Example 3 showed a normal pattern in which the leakage of FITC-dextran, which is a green dye in the retinal blood vessels, was not observed. However, in the db / db mice group , It was confirmed that the outflow of FITC-dextran appears in the gap between the blood vessels as indicated by an arrow mark. On the other hand, animal models that received 10 mg / kg of chrysin for 10 weeks showed similar vascular patterns with no FITC-dextran efflux similar to the control group.
따라서, 만성 고혈당에 노출된 제2형 당뇨병 동물 모델에 10mg/kg의 농도로 10주간 크리신을 투여한 경우에 동물 망막조직에서 나타나는 혈관 출혈을 유의적으로 억제하는 것으로 나타나, 크리신은 당뇨병성 망막병증을 억제하는 데에 뛰어난 효과를 발휘하는 것을 알 수 있다.Thus, when a model of
Claims (8)
상기 크리신은 만성 고혈당에 노출된 망막 미세 혈관 내피세포(HRMVEC)에서 비정상적인 혈관 신생에 관여하는 단백질인 저산소증 유발 인자-1α(Hypoxia-inducible factor 1-alpha, HIF-1α)와 혈관내피 성장인자(Vascular endothelial growth factor, VEGF) 생성을 감소시키는 것을 특징으로 하는 당뇨합병성 망막병증 개선용 식품 조성물.
A chrysin at a concentration of 10 to 20 [mu] M,
The above-mentioned chrysin is a protein that is involved in abnormal angiogenesis in retinal microvascular endothelial cells (HRMVEC) exposed to chronic hyperglycemia and Hypoxia-inducible factor 1-alpha (HIF-1α) and vascular endothelial growth factor endothelial growth factor (VEGF) production in a diabetic complicated retinopathy patient.
상기 식품조성물은,
만성 고혈당에 노출된 망막 미세 혈관 내피세포(HRMVEC)에서 비정상적인 혈관 신생에 관여하는 단백질인 저산소증 유발 인자-1α(Hypoxia-inducible factor 1-alpha, HIF-1α)와 혈관내피 성장인자(Vascular endothelial growth factor, VEGF) 생성을 감소시키는 것을 특징으로 하는 당뇨합병성 망막병증 개선용 식품 조성물의 제조방법.
A food composition is prepared by adding chrysin at a concentration of 10 to 20 [mu] M,
The food composition may contain,
The hypoxia-inducible factor 1-alpha (HIF-1α) and the vascular endothelial growth factor (HIF-1α), proteins involved in abnormal angiogenesis in retinal microvascular endothelial cells (HRMVEC) exposed to chronic hyperglycemia , VEGF) production of the diabetic complicated retinopathy.
상기 크리신은,
제2형 당뇨병에서 망막의 비정상적인 신생혈관(Acellular capillary) 형성을 억제하는 것을 특징으로 하는 당뇨합병성 망막병증 개선용 식품 조성물.
The method according to claim 1,
The above-
A composition for the treatment of diabetic complicated retinopathy characterized by inhibiting the formation of abnormal acellular capillaries of the retina in type 2 diabetes.
상기 크리신은,
만성 고혈당에 노출된 망막 미세 혈관 내피세포(HRMVEC)의 세포사멸을 억제하는 것을 특징으로 하는 당뇨합병성 망막병증 개선용 식품 조성물.
The method according to claim 1,
The above-
A composition for improving diabetic complications of retinopathy characterized by inhibiting apoptosis of retinal microvascular endothelial cells (HRMVEC) exposed to chronic hyperglycemia.
상기 크리신은 만성 고혈당에 노출된 망막 미세 혈관 내피세포(HRMVEC)에서 비정상적인 혈관 신생에 관여하는 단백질인 저산소증 유발 인자-1α(Hypoxia-inducible factor 1-alpha, HIF-1α)와 혈관내피 성장인자(Vascular endothelial growth factor, VEGF) 생성을 감소시키는 것을 특징으로 하는 당뇨합병성 망막병증 치료 또는 예방용 약학 조성물.
A chrysin at a concentration of 10 to 20 [mu] M,
The above-mentioned chrysin is a protein that is involved in abnormal angiogenesis in retinal microvascular endothelial cells (HRMVEC) exposed to chronic hyperglycemia and Hypoxia-inducible factor 1-alpha (HIF-1α) and vascular endothelial growth factor endothelial growth factor (VEGF) production in a patient suffering from diabetic retinopathy.
상기 약학 조성물은,
만성 고혈당에 노출된 망막 미세 혈관 내피세포(HRMVEC)에서 비정상적인 혈관 신생에 관여하는 단백질인 저산소증 유발 인자-1α(Hypoxia-inducible factor 1-alpha, HIF-1α)와 혈관내피 성장인자(Vascular endothelial growth factor, VEGF) 생성을 감소시키는 것을 특징으로 하는 당뇨합병성 망막병증 치료 또는 예방용 약학 조성물의 제조방법.
A pharmaceutical composition is prepared by adding chrysin at a concentration of 10 to 20 [mu] M,
The pharmaceutical composition may contain,
The hypoxia-inducible factor 1-alpha (HIF-1α) and the vascular endothelial growth factor (HIF-1α), proteins involved in abnormal angiogenesis in retinal microvascular endothelial cells (HRMVEC) exposed to chronic hyperglycemia , VEGF) in a patient suffering from diabetic retinopathy.
상기 크리신은,
제2형 당뇨병에서 망막의 비정상적인 신생혈관(Acellular capillary) 형성을 억제하는 것을 특징으로 하는 당뇨합병성 망막병증 치료 또는 예방용 약학 조성물.
6. The method of claim 5,
The above-
A pharmaceutical composition for the treatment or prevention of diabetic complicated retinopathy characterized by inhibiting abnormal angiogenic capillary formation of the retina in type 2 diabetes.
상기 크리신은,
만성 고혈당에 노출된 망막 미세 혈관 내피세포(HRMVEC)의 세포사멸을 억제하는 것을 특징으로 하는 당뇨합병성 망막병증 치료 또는 예방용 약학 조성물.
6. The method of claim 5,
The above-
A pharmaceutical composition for the treatment or prevention of diabetic complicated retinopathy characterized by inhibiting apoptosis of retinal microvascular endothelial cells (HRMVEC) exposed to chronic hyperglycemia.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20190093064A (en) * | 2018-01-31 | 2019-08-08 | 한림대학교 산학협력단 | Pharmaceutical composition for preventing and treating diabetic complications containing the novel chrysin derivative |
| WO2020205801A1 (en) * | 2019-03-29 | 2020-10-08 | Judith Boston | Treating ophthalmic disease using hypoxia-inducible factor inhibitors |
| CN115778937A (en) * | 2022-10-24 | 2023-03-14 | 中国人民解放军海军军医大学第一附属医院 | Application of chrysin in preparation of medicine for inhibiting retinal neovascularization |
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| JP2009527541A (en) | 2006-02-21 | 2009-07-30 | カウンシル オブ サイエンティフィク アンド インダストリアル リサーチ | Intestinal α-glucosidase inhibitor, isolation method thereof and use thereof |
| US20090325906A1 (en) * | 2008-06-27 | 2009-12-31 | Wendye Robbins | Methods and compositions for therapeutic treatment |
| KR101510257B1 (en) * | 2013-12-30 | 2015-04-09 | 한림대학교 산학협력단 | Diabetic renal fibrosis or tubulointersitial fibrosis inhibiting composition of chrysin |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009527541A (en) | 2006-02-21 | 2009-07-30 | カウンシル オブ サイエンティフィク アンド インダストリアル リサーチ | Intestinal α-glucosidase inhibitor, isolation method thereof and use thereof |
| US20090325906A1 (en) * | 2008-06-27 | 2009-12-31 | Wendye Robbins | Methods and compositions for therapeutic treatment |
| KR101510257B1 (en) * | 2013-12-30 | 2015-04-09 | 한림대학교 산학협력단 | Diabetic renal fibrosis or tubulointersitial fibrosis inhibiting composition of chrysin |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20190093064A (en) * | 2018-01-31 | 2019-08-08 | 한림대학교 산학협력단 | Pharmaceutical composition for preventing and treating diabetic complications containing the novel chrysin derivative |
| WO2019151732A1 (en) * | 2018-01-31 | 2019-08-08 | 한림대학교 산학협력단 | Pharmaceutical composition for preventing or treating diabetes complications comprising novel chrysin derivative compound as active ingredient |
| KR102032739B1 (en) | 2018-01-31 | 2019-10-16 | 한림대학교 산학협력단 | Pharmaceutical composition for preventing and treating diabetic complications containing the novel chrysin derivative |
| WO2020205801A1 (en) * | 2019-03-29 | 2020-10-08 | Judith Boston | Treating ophthalmic disease using hypoxia-inducible factor inhibitors |
| CN115778937A (en) * | 2022-10-24 | 2023-03-14 | 中国人民解放军海军军医大学第一附属医院 | Application of chrysin in preparation of medicine for inhibiting retinal neovascularization |
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