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目的利用光遗传学技术是否能有效实施光照夺获心脏,实现心脏光起搏,探讨光照强度对心脏夺获率的影响.方法 取15只C57雌性小鼠,随机分成3组:正常组6只,空载病毒组6只,Ch R2组3只(A、B、C小鼠),经颈静脉给三组小鼠分别注射100μl生理盐水,100μlp AOV-CAG-mcherry,100μlp AOV-CAG-h Ch R2(H134R)-mcherry.8周后,进行离体心脏Langendorff离体灌流,并给予473nm的蓝光光照刺激,观察小鼠的心律是否能被光刺激夺获,并比较不同功率下的光刺激对心脏夺获率的影响,观察光刺激和电刺激产生的单相动作电位(MAP)之间是否存在差异.结果 Ch R2组小鼠的心脏节律能被光照夺获,一定范围内夺获率与功率成正相关关系,且17.3m W的光照基本能满足光照夺获心律所需功率的大小,正常组和空载病毒组小鼠心脏的节律均不能被光照夺获.Ch R2组A、B、C3只小鼠光照夺获心律所需的功率大小差别较大,B小鼠心脏更容易被光夺获,可能与心脏Ch R2表达量有关.光刺激与电刺激产生的MAP存在差异,光刺激产生的MAP的达峰时间比电刺激短,且动作电位的复极时程达10%,30%,50%,70%的值小于电刺激.结论 利用光遗传学技术实现心脏光起搏技术可行.光敏蛋白Ch R2成功表达在靶器官上是光遗传学关键的一步,光刺激心脏的夺获率不仅与光照功率的大小有关,可能还与心脏表达Ch R2的量相关.
Objective To evaluate the effect of light intensity on the rate of cardiac arrest by using light-genetics technology whether light can effectively capture the heart and achieve cardiac pacing.Methods Fifteen C57 female mice were randomly divided into 3 groups: normal control group , 6 mice in the no-load virus group and 3 mice in the ChR2 group (A, B and C mice). Three groups of mice were injected with 100 μl of physiological saline, 100 μl of AOV-CAG-mcherry and 100 μl of AOV-CAG-h After 8 weeks of Ch R2 (H134R) -mcherry, isolated Langendorff hearts were perfused ex vivo and stimulated with blue light at 473 nm to observe whether the mice’s heart rhythm could be harvested by light stimulation and compared with light stimulation at different powers On the rate of cardiac seizure and observe whether there is a difference between the single phase action potential (MAP) produced by light stimulation and electrical stimulation.Results The heart rhythm of mice in Ch R2 group can be captured by light and the seizure rate within a certain range And the power has a positive correlation, and 17.3m W light can basically meet the power required to capture the heart rate of light, normal and no-load virus group mouse heart rhythm can not be captured by light.Ch R2 group A, B , C3 mouse light to capture the rhythm of the power required for the size of the larger differences, B mouse heart More likely to be captured by light, may be related to the expression of heart Ch R2.The MAP generated by light stimulation and electrical stimulation are different, the peak time of MAP generated by light stimulation is shorter than that of electrical stimulation, and the repolarization time of action potential reaches 10%, 30%, 50% and 70% were lower than that of electrical stimulation.Conclusion The technique of photoparticle photoplethysmography for cardiac pacing is feasible.The successful expression of photoprotein Ch R2 on target organs is a crucial step in optogenetics. Light The rate of capture of the stimulating heart is not only related to the amount of light power but may also be related to the amount of Ch R2 expressed in the heart.