CAJ | 학술논문

目的研究近红外线激光对视网膜神经节细胞(RGC)钾离子通道的影响.方法实验研究.将健康幼猪急性处死后取其视网膜,采用酶消化法进行猪RGC体外培养,并对细胞行免疫荧光化学染色鉴定.采用膜片钳全细胞记录模式记录原代RGC全细胞电流及经近红外线激光照射后的电流改变,绘制电流-电压曲线.将体外培养的RGC按是否接受波长为845 nm的近红外线激光照射分为激光照射组和对照组,采用t检验比较两组的平均电流峰值.结果倒置相差显微镜下观察可见,培养1周的细胞已具备典型的神经元细胞形态学特征.对培养的细胞行免疫荧光细胞化学检测,其胞体和突起呈黄绿色荧光者为阳性细胞.近红外线激光对体外培养原代RGC外向钾离子电流具有调节作用,且该调节作用呈电压依赖性.激光照射组的平均电流峰值为(634.4±86.8)Pa,与对照组的平均电流峰值(580.5±116.4)Pa比较,差异有统计学意义(n =30,t=7.923,P=0.000).结论近红外线激光作用于猪RGC可改变其外向钾离子通道特性,从而影响动作电位的形成和发放,调节神经细胞生理功能,可能有益于保护受损的神经节细胞.
목적 연구근홍외선격광대시망막신경절세포(RGC)갑리자통도적영향.방법 실험연구.장건강유저급성처사후취기시망막,채용매소화법진행저RGC체외배양,병대세포행면역형광화학염색감정.채용막편겸전세포기록모식기록원대RGC전세포전류급경근홍외선격광조사후적전류개변,회제전류-전압곡선.장체외배양적RGC안시부접수파장위845 nm적근홍외선격광조사분위격광조사조화대조조,채용t검험비교량조적평균전류봉치.결과 도치상차현미경하관찰가견,배양1주적세포이구비전형적신경원세포형태학특정.대배양적세포행면역형광세포화학검측,기포체화돌기정황록색형광자위양성세포.근홍외선격광대체외배양원대RGC외향갑리자전류구유조절작용,차해조절작용정전압의뢰성.격광조사조적평균전류봉치위(634.4±86.8)Pa,여대조조적평균전류봉치(580.5±116.4)Pa비교,차이유통계학의의(n =30,t=7.923,P=0.000).결론 근홍외선격광작용우저RGC가개변기외향갑리자통도특성,종이영향동작전위적형성화발방,조절신경세포생리공능,가능유익우보호수손적신경절세포.
Objective To investigate the influence of continuous near-infrared (CNI) laser to potassium ion channels on retinal ganglion cell ( RGC).Methods Experiment study.Porcine RGC was cultured with enzymatic digestion method in vitro by taking off the retina from the piglets.Whole-cell patch clamp mode recordings were obtained from primary cultured porcine RGC.Whole-cell currents of porcine RGC irradiated with a single-mode CNI laser of 845 nm wavelength and 30 mW power were also recorded.The primary cultured porcine RGC were divided into laser irradiation group and control group according to whether they accepted CNI laser of 845 nm wavelength.t test was used to analyze the average peak amplitude between the two groups.Results The cells had the morphological characteristics of typical neurons after one week cultured observed by inverted phase contrast microscope.The body and bumps with yellow-green fluorescence was positive cell which observed by immune cell fluorescence chemical testing.CNI laser had a regulation on outward potassium current of porcine RGC in a voltage-dependent manner.There was significant difference on the average peak amplitude of potassium current between the laser irradiation group [ (634.4 ± 86.8 ) Pa]and the control group [ ( 580.5 ± 116.4 ) Pa ],respectively ( n =30,t =7.923,P =0.000).Conclusions CNI laser can change the properties of outward K + channel.Therefore,formation and releasing of action potential is affected.Further,physiological functions of RGC are regulated,which might contribute to the protection and restoration of injured RGC.It can be provided a new scholar direction for the protection of the RGC,which are injured by glaucoma.