摘要
目的:观察艾灸听宫对膜迷路积水模型大鼠耳蜗形态、血浆血管加压素(arginine vasopressin,AVP)浓度及耳蜗水通道蛋白2(aquaporin2,AQP2)表达的影响,探讨艾灸听宫对膜迷路积水的可能调节机制。方法:将28只Wistar大鼠随机分为空白对照组,模型对照组,托伐普坦组及艾灸听宫组,每组7只。除外空白对照组,其余各组均采用腹腔注射AVP的方法复制膜迷路积水模型。空白对照组与模型对照组采取与艾灸听宫组相同的固定,不予治疗;托伐普坦组予托伐普坦溶液5 mg/kg灌胃;艾灸听宫组于听宫穴施灸20 min。各治疗组均每日治疗1次,连续治疗10 d。干预结束后,行HE染色法观察大鼠耳蜗积水程度,计算蜗管横截面积与蜗管加前庭阶横截面积之和的比值(R值);放免法检测血浆AVP浓度;免疫组化法检测耳蜗血管纹AQP2蛋白的表达水平。结果:(1)模型对照组出现膜迷路积水,前庭膜向前庭阶方向不同程度的隆起扩张,R值较空白对照组增加(P<0.01);托伐普坦组与艾灸听宫组前庭膜隆起程度均较模型对照组不同程度的减轻,R值均较模型对照组降低(P<0.01,P<0.05);艾灸听宫组R值与托伐普坦组比较,差异无统计学意义(P>0.05)。(2)模型对照组大鼠血浆AVP浓度较空白对照组升高(P<0.01);托伐普坦组与艾灸听宫组血浆AVP浓度均较模型对照组降低(均P<0.05);艾灸听宫组血浆AVP浓度与托伐普坦组比较,差异无统计学意义(P>0.05)。(3)模型对照组耳蜗血管纹AQP2表达水平较空白对照组升高(P<0.01);托伐普坦组与艾灸听宫组AQP2表达水平均较模型对照组降低(均P<0.01);艾灸听宫组AQP2表达水平与托伐普坦组比较,差异无统计学意义(P>0.05)。结论:艾灸听宫可减轻大鼠膜迷路积水,其作用机制可能与对AVP-AQP2系统的调节有关。
Objective: The objective of this study was to investigate the effects of moxibustion at Tinggong( SI19) on morphologic change,plasma arginine vasopressin( p-AVP) levels as well as expression of aquaporin2( AQP2) in rats cochlea with endolymphatic hydrops so as to explore the potential mechanism for the treatment of endolymphatic hydrops in Meniere's disease by moxibustion. Methods: Twenty-eight Wistar rats were randomly separated into a blank control group,a model control group,a tolvaptan-treated group,and a moxibustion group( n = 7). Except rats in the blank control group,the remaining animals received an intraperitoneal injection of AVP in a dose of 50 μg/( kg·d) once a day for 7 days to induce endolymphatic hydrops. In the blank control and model control groups,animals were given the same fixture as the moxibustion group once per day without other treatment. In the tolvaptan-treated group,tolvaptan solution was given transorally at a dose of 5 mg/kg once a day during 10 days. Animals in the moxibustion group were administered moxibustion at Tinggong( SI19) for 20 minutes once per day for 10 days. The severity of cochlear hydrops was evaluated by hematoxylin-eosin( HE) staining and then the ratio of scala media( SM) area to SM + scala vestibuli( SV)area( R value) was calculated. Plasma levels of AVP were determined by radioimmunoassay( RIA). The expression ofAQP2 in the stria vascularis was observed by immunohistochemical methods. Results:(1)The histologic morphometric results showed that the Reissner's membrane was extended markedly and bulged into SV in cochleae of the model control group and endolymphatic hydrops was noted. Statistical analysis revealed that R value in the model control group showed a significant increase compared with that in the blank control group( P < 0. 01). The distension of Reissner's membrane was less obvious in the tolvaptan-treated group and the moxibustion group when compared with the model control group.R value in the tolvaptan-treated group and the moxibustion group was significantly less than that in model control group( P < 0. 01,P < 0. 05). The degree of the hydrops in the moxibustion group was not different from that in the tolvaptan-treated group( P > 0. 05).(2) He p-AVP concentrations in the model control group were higher than that of the blank control group( P < 0. 01). The p-AVP concentrations in the tolvaptan-treated group and the moxibustion group were all lower than that of the model control group( all P < 0. 05). The p-AVP concentrations in the moxibustion group were not different from that in the tolvaptan-treated group( P > 0. 05).(3)The AQP2 protein expression in the stria vascularis in rats of model control group was significantly increased when compared with the blank control group( P < 0. 01). The AQP2 protein expression in the stria vascularis in rats of tolvaptan-treated group and moxibustion group were all lower than that of the model control group( all P < 0. 01). The AQP2 protein expression in the stria vascularis in rats of moxibustion group was not different from that in the tolvaptan-treated group( P > 0. 05). Conclusions: These results indicate that moxibustion stimulation at Tinggong( SI19) could reduce cochlear hydrops in rats with endolymphatic hydrops,and the water homeostasis in the inner ear is likely regulated by moxibustion via the AVP-AQP2 system.
引文
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