【摘要】：目的：研究小鼠左心室肌細(xì)胞收縮與鈣瞬變對(duì)頻率和氧化壓力應(yīng)答的跨膜梯度。材料：6周鼠齡的C57雄性小鼠左心室內(nèi)／外膜層肌細(xì)胞分離采用Langendorff灌流術(shù)。心肌細(xì)胞鈣瞬變和收縮舒張功能同步檢測(cè)使用的是雙激光熒光光電倍增系統(tǒng)。結(jié)果：實(shí)驗(yàn)統(tǒng)計(jì)了來自6只小鼠的22個(gè)內(nèi)膜心肌細(xì)胞和26個(gè)外膜心肌細(xì)胞,發(fā)現(xiàn)在刺激頻率為lhz,臺(tái)式液灌流情況下,正常小鼠左心室內(nèi)膜心肌細(xì)胞的舒張期鈣濃度為0.521±0.023(F340/380),大于外膜心肌細(xì)胞的0.466±0.022(F340/380)(p0.05)；鈣瞬變振幅為0.241±0.014(F340/380)大于外膜心肌細(xì)胞的0.203±0.011(F340/380)(p0.05)；肌織網(wǎng)鈣釋放速率為20.89±0.923(F/s)小于外膜心肌細(xì)胞的22.823±0.814 F/s(p0.05)；肌織網(wǎng)鈣回收速率為2.459±0.292(F/s),小于外膜心肌細(xì)胞的3.267±0.245(F/s)(p0.05)。同時(shí),內(nèi)膜心肌細(xì)胞的肌小節(jié)長(zhǎng)度為1.816±0.033(um)小于外膜心肌細(xì)胞的1.834±0.011(um)(p0.05)；肌小節(jié)收縮振幅為0.034±0.005(um)小于外膜心肌細(xì)胞的0.023±0.002(um)(p0.05)。刺激頻率增加后,內(nèi)膜心肌細(xì)胞的舒張期鈣濃度增加了16.29%±2.64%,大于外膜心肌細(xì)胞的8.91%±1.12%(p0.05)；收縮期鈣瞬變振幅增加了32.12%±5.69%,大于外膜心肌細(xì)胞的14.12%±1.67%(p0.05)；胞質(zhì)鈣回收速率的增加129.29%±24.72%,大于外膜心肌細(xì)胞的55.50%±4.55%(p0.05)。相應(yīng)地,肌小節(jié)收縮振幅增加了250.30%±42.08%(n=12)大于外膜層心肌細(xì)胞的49.97%±7.56%(n=23,P=0.020.05)。肌小節(jié)縮短的最大速率加速了433.15%±71.25%(n=12)大于外膜層心肌細(xì)胞75.74%±13.95%(n=23,P=0.0090.05)。當(dāng)給予100 uM H202暴露1min使內(nèi)膜心肌細(xì)胞收縮振幅增加了0.0155±0.0017(um)大于外膜心肌細(xì)胞的0.0096±0.0009(um),P0.05。內(nèi)膜心肌細(xì)胞鈣瞬變振幅增加了0.0737±0.0043(F340/380)大于外膜心肌細(xì)胞的0.0284±0.0014(F340/380),P0.05。正常小鼠內(nèi)外膜心肌細(xì)胞的鈣瞬變振幅存在0.045±0.006(F340/380)的差異,而心衰小鼠內(nèi)外膜心肌細(xì)胞的鈣瞬變差異為0.005±0.001(F340/380)(P0.05)結(jié)論：小鼠左心室內(nèi)膜心肌細(xì)胞鈣瞬變振幅和收縮力均強(qiáng)于外膜心肌細(xì)胞。內(nèi)膜層心肌細(xì)胞鈣瞬變和收縮舒張慢于外膜層心肌細(xì)胞。內(nèi)膜層心肌細(xì)胞鈣瞬變和收縮舒張對(duì)頻率的反應(yīng)較為膜層心肌細(xì)胞更敏感。低濃度段時(shí)間的氧化壓力使內(nèi)膜心肌細(xì)胞收縮和鈣瞬變代償性增加大于外膜心肌細(xì)胞。心衰小鼠內(nèi)外膜心肌細(xì)胞鈣瞬變的跨膜梯度消失。
[Abstract]:Aim: to study the transmembrane gradient of contractile and calcium transient responses to frequency and oxidative pressure in mouse left ventricular myocytes. Materials: the left ventricular / outer lamellar myocytes of 6 week old C57 male mice were isolated and perfused with Langendorff. Double-laser fluorescence photomultiplier system is used for simultaneous detection of calcium transient and systolic-diastolic function of myocardial cells. Results: 22 endomyocytes and 26 adventitia cardiomyocytes from 6 mice were counted. The results showed that the stimulation frequency was lhz, table perfusion. The diastolic calcium concentration of left ventricular endomyocytes in normal mice was 0.521 鹵0.023 (F340 / 380), which was higher than that of adventitia cardiomyocytes (0.466 鹵0.022) (p0.05). The transient amplitude of calcium was 0.241 鹵0.014 (F340 / 380) and 0.203 鹵0.011 (F340 / 380) in epicardial cardiomyocytes (p0.05). The calcium release rate of muscle mesh was 20.89 鹵0.923 (F / s) less than that of 22.823 鹵0.814 F / s of epicardial cardiomyocytes (p0.05). The calcium recovery rate of muscle netting was 2.459 鹵0.292 (F / s), which was lower than 3.267 鹵0.245 (F / s) of epicardial cardiomyocytes (p0.05). At the same time, the length of muscle section of endomyocytes was 1.816 鹵0.033 (um), which was smaller than that of adventitia cardiomyocytes (1.834 鹵0.011 (um) (, p0.05). The contractile amplitude of muscle section was 0.034 鹵0.005 (um), which was lower than that of adventitia cardiomyocytes (0.023 鹵0.002 (um) (, p0.05). After stimulation, the diastolic calcium concentration increased by 16.29% 鹵2.64%, which was higher than that of adventitia cardiomyocytes (8.91% 鹵1.12%, p0.05). The amplitude of transient calcium in systolic phase increased by 32.12% 鹵5.69%, which was higher than that of adventitia cardiomyocytes by 14.12% 鹵1.67% (p0.05). The recovery rate of calcium in cytoplasm was increased by 129.29% 鹵24.72%, which was higher than that of adventitia cardiomyocytes by 55.50% 鹵4.55% (p0.05). Correspondingly, the contractile amplitude of muscle section was increased by 250.30% 鹵42.08% (nm12) than that of adventitia cardiomyocytes (49.97% 鹵7.56%). The maximal rate of shortening of muscle section was increased by 433.15% 鹵71.25% (nm12) than that of adventitia cardiomyocytes (75.74% 鹵13.95%). When exposed to 100 uM H202, the contractile amplitude of endomyocytes was increased by 0.0155 鹵0.0017 (um) than by 0.0096 鹵0.0009 (um), P0.05 of epicardial cardiomyocytes. The amplitude of calcium transient was increased by 0.0737 鹵0.0043 (F340 / 380) in endomyocytes than 0.0284 鹵0.0014 (F340 / 380) in adventitial cardiomyocytes (P0.05). In normal mice, the amplitude of calcium transient was 0.045 鹵0.006 (F340 / 380). The difference of calcium transient was 0.005 鹵0.001 (F340 / 380) (P0.05). Conclusion: the amplitude and contractility of calcium transient in mouse left ventricular endomyocytes are stronger than that of adventitia cardiomyocytes. Calcium transient and diastolic activity of endometrial myocardial cells was slower than that of adventitia cardiomyocytes. Calcium transient and contractile relaxation were more sensitive to frequency than membranous myocardial cells. The contractility and calcium transient compensability of myocardial cells were increased by oxidative pressure at low concentration than that of adventitia cardiomyocytes. The transmembrane gradient of calcium transient in cardiac myocytes of heart failure mice was disappeared.
【學(xué)位授予單位】：武漢大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R54

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