目舒丸对兔眼睫状体平滑肌细胞内舒缩传导信号的影响  

Effect of Mushu Pill on the signal transduction of relaxation and contraction in rabbit ciliary body smooth muscle cells

作　　者：宿蕾艳[1] 张莎莎 张明明[1] 魏春秀[1] 方素萍[2] 王颖[1] 胡瑛[1] 庄曾渊[1] SU Leiyan;ZHANG Shasha;ZHANG Mingming(Eye Hospital,China Academy of Chinese Medical Sciences,Beijing 100040,China)

机构地区：[1]中国中医科学院眼科医院,北京100040 [2]中国中医科学院广安门医院,北京100053

出　　处：《中国中医眼科杂志》2021年第12期846-850,855,共6页China Journal of Chinese Ophthalmology

基　　金：国家自然科学基金项目(81303008)。

摘　　要：目的观察目舒丸对兔眼睫状体平滑肌细胞内钙依赖性及非钙依赖性舒缩信号传导的影响。方法分离提取原代新西兰大白兔眼睫状体平滑肌细胞均匀接种于孔板中,随机选出27个接种孔,并随机分为9组,每组3个孔,分别命名为A组~I组。其中A~G组分别予8、4、2、1、0.5、0.25、0.125 mg/mL 7个不同浓度目舒丸进行干预,在不同干预时间点观察目舒丸对兔眼睫状体平滑肌细胞舒缩作用,确定最佳干预时间及最佳药物浓度。H组予0.5 mmol/L氯化乙酰胆碱培养,I组先予1.0 mg/mL目舒丸溶液培养,再予相同浓度的氯化乙酰胆培养,检测2组细胞内蛋白激酶C(PKC)、同源基因家族成员A(RhoA)的基因表达,观察目舒丸对睫状体平滑肌细胞内与舒缩有关的钙依赖性及非钙依赖性信号传导的影响。结果(1)细胞直径:与给药前比较,A组给药后所有时间点细胞直径均缩短;C组给药后4、10 min细胞直径均伸长;D组给药后1、2、4、10 min细胞直径均伸长;E组给药后2、4、10 min细胞直径均伸长,以上比较均有统计学意义(P<0.05)。组间比较,与A组给药后各时间点比较,B~G组细胞直径均伸长;与B组给药后10 min比较,E组细胞直径缩短;与C组给药后0.5 min比较,E组细胞直径缩短;与C组给药后1 min比较,D、G组细胞直径伸长;与D给药后10 min比较,E组细胞直径伸长;与E组给药后0.5 min比较,F组细胞直径伸长;与F组给药后2 min比较,G组细胞直径伸长,以上差异均有统计学意义(P<0.05)。(2)PKC及RhoA的表达:与H组比较,I组的PKC及RhoA表达量降低,差异均有统计学意义(t_(PKC)=-3.566,P=0.023;t_(RhoA)=-8.895,P=0.001)。结论目舒丸能影响睫状肌舒缩,并呈一定的时间依赖关系,其中1.0 mg/mL的浓度作用最强,可能与干预兔眼睫状体平滑肌细胞钙依赖性及非钙依赖性信号因子有关。OBJECTIVE To observe the effect of Mushu Pill on calcium-dependent and non-calcium-dependent relaxation and contraction signal transduction in ciliary body smooth muscle cells of rabbits.METHODS New Zealand white rabbits'primary ciliary body smooth muscle cells were isolated and evenly inoculated into the well plates.A total of 27 inoculation wells were randomly selected and randomly divided into nine groups with three wells in each group,named groups A~I,respectively.Group A~G were intervened with 7 different concentrations of Mushu Pill(8,4,2,1,0.5,0.25 and 0.125 mg/mL),respectively.The effect of Mushu Pill on the relaxation and contraction of ciliary smooth muscle cells were observed at different intervention time points to determine the best intervention time and concentration of Mushu Pill.Group H was cultured with 0.5 mmol/L acetylcholine chloride,while group I was first cultured with 1.0 mg/mL Mushu Pill solution and then cultured with the same concentration of acetylcholine chloride.The gene expressions of PKC and RhoA in the H and I cells groups were detected,and the effects of Mushu Pill on calcium-dependent and non-calcium-dependent signals transduction related to relaxation and contraction in ciliary smooth muscle cells were observed.RESULTS(1)Cell diameter:Compared with that before administration,the cell diameter of group A shortened at all time points after Mushu pill administration.In group C,the cell diameter elongated at 4 and 10 min after administration.In group D,the cell diameter elongated at 1,2,4,and 10 min after administration.In group E,the cell diameter elongated at 2,4,and 10 min after administration,and the differences were all statistically significant(P<0.05).Compared with group A at each time point after administration,the cell diameters of group B~G elongated.Compared with group B at 10 min after administration,the cell diameter in Group E shortened.Compared with group C at 0.5 min after administration,the cell diameter in group E shortened,and the cell diameter in groups D and G e

关 键 词：目舒丸 睫状体平滑肌 视疲劳 

分 类 号：R285.5[医药卫生—中药学]