CAJ | 학술논문

脊椎动物中,非晶状体βγ-晶状体蛋白广泛分布于各种组织,但是功能知之甚少.三叶因子在创伤修复与肿瘤发生中具有重要作用,其分子作用机制尚不清楚.非晶状体βγ晶状体蛋白与三叶因子蛋白复合物(βγ-CAT)是一个从大蹼铃蟾皮肤分泌物中分离的一类全新的蛋白复合物.研究表明,βγ-CAT能够诱导离体的兔胸主动脉产生快速而持续的收缩,结合药理学抑制剂,细胞培养,激光共聚焦显微镜和免疫荧光原位组化,从细胞和分子水平对其作用机制进行研究.结果表明:.βγ-CAT诱导兔胸主动脉产生的收缩效应为剂量依赖(2-35 nmol/L)和内皮依赖(P＜0.01).在βγ-CAT(25nmol/L)处理的主动脉环的内皮细胞层检测到肿瘤坏死因子-α的释放.同时,βγ-CAT能够诱导原代培养的兔胸主动脉内皮细胞(RAEC)快速释放肿瘤坏死因子-α,βγ-CAT(25nmol/L)分别处理5和30min,RAEC释放的肿瘤坏死因子-α的浓度分别为(34.17±5.10)pg/mL和(98.01±4.67)pg/mL(P＜0.01).表明肿瘤坏死因子-α在βγ-CAT诱导兔胸丰动脉产生的收缩效应中发挥重要作用.为进一步深入研究非晶状体βγ晶状体蛋白与三叶因子的生理功能提供了新的思路和线索.
척추동물중,비정상체βγ-정상체단백엄범분포우각충조직,단시공능지지심소.삼협인자재창상수복여종류발생중구유중요작용,기분자작용궤제상불청초.비정상체βγ정상체단백여삼협인자단백복합물(βγ-CAT)시일개종대복령섬피부분비물중분리적일류전신적단백복합물.연구표명,βγ-CAT능구유도리체적토흉주동맥산생쾌속이지속적수축,결합약이학억제제,세포배양,격광공취초현미경화면역형광원위조화,종세포화분자수평대기작용궤제진행연구.결과표명:.βγ-CAT유도토흉주동맥산생적수축효응위제량의뢰(2-35 nmol/L)화내피의뢰(P＜0.01).재βγ-CAT(25nmol/L)처리적주동맥배적내피세포층검측도종류배사인자-α적석방.동시,βγ-CAT능구유도원대배양적토흉주동맥내피세포(RAEC)쾌속석방종류배사인자-α,βγ-CAT(25nmol/L)분별처리5화30min,RAEC석방적종류배사인자-α적농도분별위(34.17±5.10)pg/mL화(98.01±4.67)pg/mL(P＜0.01).표명종류배사인자-α재βγ-CAT유도토흉봉동맥산생적수축효응중발휘중요작용.위진일보심입연구비정상체βγ정상체단백여삼협인자적생리공능제공료신적사로화선색.
In vertebrates, non-lens βγ-crystallins are widely expressed in various tissues and their functions are not well known.The molecular mechanisms of trefoil factors (TFFs), which involved in mucosal healing and tumorigenesis, have remained elusive.βγ-CAT is a novel multifunctional protein complex of non-lens βγ-crystallin and trefoil factor from frog skin secretions. Here we report that βγ-CAT could induce sustained contraction of isolated rabbit aortic tings in dosage (2-35 nmol/L) and endothelium dependent manners (P＜0.01). In addition, in situ immunofluorescence indicated that positive TNF-α signals were mainly detected at the endothelial cell layer of βγ-CAT (25nmol/L) treated rings. Furthermore, βγ-CAT induced primary cultured rabbit thoracic aortic endothelial cells (RAECs) rapidly to release TNF-α. After βγ-CAT (25nmol/L) treated for 10 and 30 min, the levels of the endothelial cells released TNF-α were 34.17±5.10 pg/mL and 98.01±4.67 pg/mL (P＜0.01), respectively. In conclusion, βγ-CAT could induce sustained contraction of isolated aortic rings, and the contractile effect might be partially explained by the release of TNF-α. These findings will give new insight into understanding the functions and physiological roles of non-lens βγ-crystallins and trefoil factors.