本文選題：TRPV4 + 膜片鉗��； 參考：《華中科技大學(xué)》2006年碩士論文

【摘要】： TRPV4是作為低滲透壓激活的滲透壓感受器被發(fā)現(xiàn)的。Phorbol ester 4a-phorbol 12,13-didecanoate(4аPDD)是一種合成佛波脂,它是一種非蛋白磷脂酶C途徑激活通道的佛波脂類,對TRPV4有強(qiáng)效的特異性的激動(dòng)作用。滲透壓和佛波脂兩者的通道激活機(jī)制還未曾清楚�，F(xiàn)已明確TRPV4是一種非選擇性的外向整流陽離子通道,對鈣離子和鈉離子都有通透性,鈣鈉的通透比值為5~7。TRPV4受體激動(dòng)可以引起細(xì)胞內(nèi)鈣離子濃度的增加,對鈉離子的影響還未見相關(guān)文獻(xiàn)報(bào)道。本實(shí)驗(yàn)擬用低滲和4аPDD刺激TRPV4來觀察鈉電流的變化。 (1)應(yīng)用全細(xì)胞膜片鉗技術(shù),在培養(yǎng)的大鼠三叉神經(jīng)節(jié)神經(jīng)元上記錄低滲外液對細(xì)胞膜電壓依賴性鈉電流的影響。結(jié)果表明:低滲外液可濃度依賴性地抑制電壓依賴性鈉電流峰值。280mOsm,260mOsm,220mOsm分別抑制三叉神經(jīng)節(jié)神經(jīng)元電壓依賴性鈉電流峰值為24.2%±10.2% (n=6, P 0.05)和45.7%±15.6% (n=8, P 0.05)以及70.5%±18.1% (n=6, P 0.05),IC50為268mOsm。300mOsm,280mOsm,260mOsm,220mOsm對電壓依賴性鈉電流的翻轉(zhuǎn)電位分別為28.0mV、22.7 mV、17.4mV、7.3 mV (n=6, P 0.05)。低滲外液對電壓依賴性鈉電流激活曲線無明顯影響;左移失活曲線。300mOsm、280mOsm、260mOsm、220mOsm失活曲線擬合的半數(shù)失活電壓,曲線的斜率因子分別為V1/2=-45.8±9.6mV,k=-17.54±6.9;V1/2=-47.2±7.8mV, k=-17.1±6.0 ;V1/2=-56.4±11.2mV, k=-14.4±5.8;V1/2=-60.6±10.8mV, k=-11.3±4.2 (n=6, P 0.05)。 (2)應(yīng)用全細(xì)胞膜片鉗技術(shù),在培養(yǎng)的大鼠三叉神經(jīng)節(jié)神經(jīng)元上記錄佛波脂4alphaPDD對細(xì)胞膜電壓依賴性鈉電流的影響。佛波脂4alphaPDD也可濃度依賴性地抑制大鼠三叉神經(jīng)節(jié)神經(jīng)元電壓依賴性鈉電流峰值。0.1,0.3,1,3,10μmol·L-1 4аPDD分別抑制三叉神經(jīng)節(jié)神經(jīng)元電壓依賴性鈉電流峰值24.68±7.2%(n=8, P 0.05),32.02±9.4%(n=7, P 0.05),52.7±12.2%(n=6, P 0.05),67.48±9.8%(n=7, P 0.05),73.9±14.6%(n=6, P 0.05),IC50為0.41μmol·L-1。0.1μmol·L-1 4аPDD對電壓依賴性鈉電流的電壓依賴性、激活曲線、失活曲線均無明顯影響。
[Abstract]:Phorbol ester 4a-phorbol 1213-didecanoate 4 (PDD) is a kind of synthetic phorbol lipids, which is a nonprotein phospholipase C pathway activator of phorbol lipids, which has a strong and specific activation effect on TRPV4.The mechanism of osmotic pressure and phorbol fat channel activation is not clear.It has been established that TRPV4 is a non-selective outward rectifier cation channel, which has permeability to both calcium and sodium ions. The permeability ratio of calcium and sodium is that the activation of 5~7.TRPV4 receptor can cause the increase of intracellular calcium ion concentration.The effect on sodium ion has not been reported.In this experiment, hypotonic and 4 PDD stimulation of TRPV4 were used to observe the changes of sodium current.1) the effects of hypotonic solution on the voltage-dependent sodium currents of rat trigeminal ganglion neurons were recorded by whole-cell patch clamp technique.The results showed that hypotonic solution could inhibit the peak value of voltage-dependent sodium current in a concentration-dependent manner. 280 m Osmon 260 m Osmon 220 mOsm in trigeminal ganglion neurons (24.2% 鹵10.2%, P 0.05) and 45.7% 鹵15.6% n8, P 0.05), and 70.5% 鹵18.1% P0.05% IC50 in trigeminal ganglion neurons, respectively.The flipping potentials of the voltage-dependent sodium current for 280mOsm.300mOsm-280mOsm-220mOsm were 28.0mV ~ 22.7mV ~ 17.4mV ~ (7.3) MV ~ 7.3mV / n ~ (6), P = 0.05 ~ (-1), respectively.浣庢笚澶栨恫瀵圭數(shù)鍘嬩緷璧栨，

本文編號(hào)：1761589