Mechanical Stretchが平滑筋細胞IP_3受容体発現量に及ぼす影響

机械拉伸对平滑肌细胞IP_3受体表达水平的影响

• 批准号: 08770526
• 负责人: 山本 寿郎
• 金额: $ 0.64万
• 依托单位: Hyogo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Encouragement of Young Scientists (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-08770526/
• 关键词: mechanical stretch; IP_3受容体; 血管平滑筋

【目的】Inositol 1,4,5-trisphosphate(IP_3)は,種々のsignal伝達の重要なsecond messengerである。またIP_3受容体に結合し,そのIP_3受容体は細胞内storeより|Ca^<2+>|放出を引き起こす。これらは平滑筋細胞の収縮や肥大に関与していると考えられている。mechanical stretchは平滑筋細胞の肥大を引き起こすが,平滑筋細胞へのmechanical stretchがIP_3受容体の発現量に影響を与えるかを検討した報告は少ない。本研究はこの点を明らかにするために平滑筋細胞のmechanical stretchによるIP_3受容体発現量の変化を検討した。【方法】ヒト臍帯静脈より平滑筋細胞を培養,type1,2,3IP_3受容体抗体を用いWestern blot法にて各subtypeの蛋白質発現量を比較した。またflexible bottomed culture plate上に平滑筋細胞を培養,Flexercell Strain Unitを用い40k cycle,1hz 20% elongationでmechanical stretchを行い,RT-PCR法にてtype2IP_3受容体mRNA量,type2IP_3受容体抗体を用いWestern blot法にてtype2IP_3受容体蛋白質量の変化を検討した。【結果】血管平滑筋ではtype2IP_3受容体蛋白質が最も多く存在し,type3IP_3受容体蛋白質が最も少なかった。mechanical stretchによるtype2IP_3受容体mRNA量は40k cycleで約60%増加,蛋白質は40k cycleで約30%増加した。【総括】type1,2,3IP_3受容体蛋白質量は小脳などの神経組織と異なる比率を示し,これが各臓器の|Ca^<2+>|動態の差異に関与している可能性が示唆された。mechanical stretchがIP_3受容体mRNA量のみならず,蛋白質量も増加する事よりIP_3受容体は平滑筋細胞の肥大に関与する事が示唆された。これらより本研究は当初の見込みを達成できたと思われるが,今後とも継続的な検討が必要である。

[objective] Inositol 1, 4, 5, Inositol trisphosphate (IP_3), a kind of signal, is the most important one in second messenger. When the IP_3 acceptor is combined, the IP _ 3 acceptor is loaded with intracellular store depletion | Ca ^ & lt;2+> | the release induces the release of the receptor. The smooth gluten cells are enlarged and the smooth tendons are not smooth. Mechanical stretch smooth gluten cell hypertrophy leads to mechanical stretchy IP _ 3 receptacle, which affects the volume of the cell and reports less. The purpose of this study is to determine the size of the smooth muscle cells, mechanical stretch, IP _ 3 receptacles, volume and volume. [methods] the smooth muscle cells were cultured and the type1,2,3IP_3 receptor antibodies were measured by Western blot method. The subtype proteins were measured by Elisa. The smooth gluten cells on the flexible bottomed culture plate were cultured, and the Flexercell Strain Unit was measured with 40k cycle,1hz 20% elongation mechanical stretch. The mRNA content of type2IP_3 receptor and the antibody of type2IP_3 receptor were measured by RT-PCR method. Type2IP_3 receptor protein was measured by Western blot method. [results] the expression of vascular smooth muscle type 2IP _ 3 receptor protein was the most frequent, while that of type3IP_3 receptor protein was the least. The amount of mRNA of mechanical stretch receptor type 2IP _ 3 is about 60% of that of 40k cycle, and that of protein of 40k cycle is about 30%. [including] type1,2,3IP_3 acceptor protein levels are small, and tissue activity ratios are displayed, and each device is used | Ca ^ & lt;2+> | dynamic response is poor, and the possibility is different. The amount of mRNAs of mechanical receptor IP _ 3, the amount of protein, the amount of protein, the amount of smooth muscle, the cell hypertrophy of IP_3 receptor, and the amount of protein, the amount of protein, the Since the beginning of this study, I have thought about it, and now it is necessary for me to do so.