史莹莺,朱敏,叶恬恬,等.电针干预豚鼠膜迷路积水的参数优选及其对耳蜗AQP2表达的影响[J].浙江中医药大学学报,2020,44(2):133-139. |
电针干预豚鼠膜迷路积水的参数优选及其对耳蜗AQP2表达的影响 |
Parametric Optimization of Electroacupuncture Against Endolymphatic Hydrops in Guinea Pigs and Its Effects on Cochlea AQP2 Expression |
DOI:10.16466/j.issn1005-5509.2020.02.005 |
中文关键词: 膜迷路积水 电针 频率 耳蜗形态 水通道蛋白2 |
英文关键词: endolymphatic hydrops electroacupuncture frequency cochlear morphology AQP2 |
基金项目:国家自然科学基金项目(81704153) |
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中文摘要: |
[目的]观察不同频率电针对豚鼠膜迷路积水的干预情况,以期优选出电针治疗膜迷路积水的适宜治疗参数,并初步探讨电针治疗梅尼埃病的可能机制。[方法]健康雄性豚鼠48只,采用完全随机法分为空白组、模型组、假电针组、电针组(据不同频率分为3个亚组),每组8只。除空白组外,其余各组均通过腹腔注射醋酸去氨加压素的方法建立膜迷路积水模型。空白组不进行干预;模型组造模后仅采取与各治疗组动物相同的固定方式,不行其他干预;电针组中各个亚组,取“百会”和左侧“听宫”穴,分别以2、15、100Hz 3种不同频率进行电针治疗,1次/d,输出电流1mA,留针20min;假电针组予针刺“百会”和左侧“听宫”穴,但不通电,其余处理同电针组,均连续治疗10d。干预结束后,采用听觉脑干诱发电位仪检测各组豚鼠听性脑干反射(auditory brairstem response, ABR)反应阈值,HE染色检测耳蜗积水程度,并通过公式蜗管横截面积/(蜗管横截面积+前庭阶横截面积)计算出比值(R值),免疫组化染色法检测耳蜗水通道蛋白2(aquaporin 2,AQP2)表达情况。[结果]各组豚鼠ABR反应阈值比较,差异无统计学意义(P>0.05)。与空白组比较,模型组豚鼠耳蜗R值增加(P<0.01);与模型组比较,各治疗组豚鼠耳蜗R值均降低(P<0.01,P<0.05);与假电针组比较,100Hz电针组R值降低(P<0.01),2、15Hz电针组R值均较假电针组降低,但差异均无统计学意义(P>0.05)。各电针组组间比较,100Hz电针组R值较2、15Hz电针组降低,差异有统计学意义(均P<0.05);2、15Hz电针组比较差异无统计学意义(P>0.05)。与空白组比较,模型组豚鼠耳蜗AQP2表达增强(P<0.01);与模型组比较,各治疗组豚鼠耳蜗AQP2表达减弱(P<0.01,P<0.05);各治疗组间比较AQP2表达差异无统计学意义(P>0.05)。[结论]假电针及电针治疗均能减轻豚鼠膜迷路积水,其中100Hz电针效果最佳,其作用机制可能与下调耳蜗AQP2的表达有关。 |
英文摘要: |
[Objective] To observe the intervention of electroacupuncture (EA) of different current frequencies against endolymphatic hydrops in guinea pigs, so as to optimize treatment parameters of EA against endolymphatic hydrops; and explore the possible mechanism of EA against Meniere's disease. [Methods] Forty-eight healthy male guinea pigs were randomly divided into blank group, model group, sham EA group and EA group(including 3 subgroups according to different frequencies), with eight guinea pigs in each. All guinea pigs except blank group were injected with arginine vasopressin to establish endolymphatic hydrops model. No treatment was given to guinea pigs in blank group; guinea pigs in model group were fixed with the same method as treatment groups without other intervention; guinea pigs in EA groups were treated with EA at "Baihui"(GV 20) and unilateral "Tinggong" (SI 19) with 3 different current frequencies(2Hz,15Hz,100Hz), once a day, current intensity for 20 mins per treatment; and sham EA group received usual acupuncture on the same acupoints at the same times. All the treatment was given for 10 days. After treatment, auditory brainstem response(ABR) thresholds of guinea pigs in each group were recorded to evaluate the hearing changes. The severity of cochlear hydrops was evaluated by hematoxylin-eosin (HE) staining, and then the R value was calculated, R value meant ratio of scala media(SM) area to SM plus scala vestibuli(SV) area. The expression of measuring cochlea aquaporin 2(AQP2) in the cochlea was observed by immunohistochemical methods. [Results] There was significant differences in average ABR thresholds in each group(P>0.05). Compared with blank group, R value of model group increased(P<0.01). Compared with model group, R value of EA subgroups and sham EA group decreased(P<0.01, P<0.05). Compared with sham EA group, R value in 100Hz EA group decreased(P<0.01); R value in 2Hz and 15Hz EA group was lower than sham EA group, but there was no significant differences(P>0.05). R value in 100Hz EA group was significantly lower than that in 2Hz and 15Hz EA group(all P<0.05), while there was no difference in R value between 2Hz and 15 Hz EA group (P>0.05). Compared with blank group, the expression of AQP2 in cochlea of model group increased significantly(P<0.01); compared with model group, the expression of AQP2 in cochlea of treatment groups decreased(P<0.01, P<0.05). There were no differences in expression of AQP2 among the treatment groups(P>0.05). [Conclusion] Both sham EA and EA treatment could suppress the development of endolymphatic hydrops, 100Hz EA is the most effective in this experiment, and the mechanism is likely to be related with down-regulating the expression of AQP2 in cochlea. |
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