艾灸听宫对大鼠膜迷路积水的影响
|
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.
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
[1]卫旭东.梅尼埃病检查方法研究进展[J].临床耳鼻咽喉头颈外科杂志,2015,29(5):385-388.
[2]Hallpike C S,Cairns H.Observations on the Pathology of Meniere's Syndrome:(Section of Otology)[J].Proc R Soc Med,1938,31(11):1317-1336.
[3]张大帅,穆帅,刘颖,等.非肽类精氨酸加压素受体拮抗剂的研究进展[J].中国药物化学杂志,2014,24(2):103-115.
[4]Kakigi A,Takeda T.Antidiuretic hormone and osmolality in patients with Meniere's disease[J].ORL J Otorhinolaryngol Relat Spec,2009,71(1):11-13.
[5]Sawada S,Takeda T,Saito H.Antidiuretic hormone and psychosomatic aspects in Meniere's disease[J].Acta Otolaryngol Suppl,1997,528:109-112.
[6]Kitano H,Takeda T,Takeda S,et al.Endolymphatic hydrops by administration of vasopressin in the rat[J].Acta Histochem,Cytoc,2001,34(4):229-233.
[7]Gu F M,Han H L,Zhang L S.Effects of vasopressin on gene expression in rat inner ear[J].Hear Res,2006,222(1-2):70-78.
[8]Nishimoto G,Zelenina M,Li D,et al.Arginine vasopressin stimulates phosphorylation of aquaporin-2 in rat renal tissue[J].Am J Physiol,1999,276(2 Pt 2):F254-F259.
[9]Kumagami H,Loewenheim H,Beitz E,et al.The effect of anti-diuretic hormone on the endolymphatic sac of the inner ear[J].Pflugers Arch,1998,436(6):970-975.
[10]Long A F,Xing M,Morgan K,et al.Exploring the Evidence Base for Acupuncture in the Treatment of Ménière's Syndrome—A Systematic Review[J].Evidence-Based Complementary and Alternative Medicine,2011,2011:1-13.
[11]蒋丽元,倪芳英,王灿军,等.不同针灸方法对膜迷路积水模型豚鼠听力及耳蜗形态结构的影响[J].中国比较医学杂志,2015,25(2):22-25,33.
[12]Takeda T,Taguchi D.Aquaporins as potential drug targets for Meniere's disease and its related diseases[J].Handb Exp Pharmacol,2009(190):171-184.
[13]马海珠,钟时勋,温雅.醋酸去氨加压素对豚鼠耳蜗水通道蛋白2表达的调节作用[J].听力学及言语疾病杂志,2014,22(3):286-289.
[14]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:314-329.
[15]中国中医研究院针灸研究所.标准针灸经穴部位图谱[M].北京:中国标准出版社,1990:54,84.
[16]Naganuma H,Kawahara K,Tokumasu K,et al.Effects of arginine vasopressin on auditory brainstem response and cochlear morphology in rats[J].Auris Nasus Larynx,2014,41(3):249-254.
[17]黄菁.苓桂术甘汤合泽泻汤对豚鼠膜迷路积水模型小肠AQP8表达影响的研究[D].成都:成都中医药大学,2011.
[18]Takeda T,Takeda S,Kitano H,et al.Endolymphatic hydrops induced by chronic administration of vasopressin[J].Hear Res,2000,140(1-2):1-6.
[19]Takumida M,Kakigi A,Egami N,et al.Localization of aquaporins 1,2,and 3 and vasopressin type 2 receptor in the mouse inner ear[J].Acta Otolaryngol,2012,132(8):807-813.
[20]陈婷,张榕.水通道蛋白2在内淋巴囊上皮和肾脏中表达的研究[J].听力学及言语疾病杂志,2008,16(5):405-409.
[21]Sawada S,Takeda T,Kitano H,et al.Aquaporin-2 regulation by vasopressin in the rat inner ear[J].Neuroreport,2002,13(9):1127-1129.
[22]周道高.灸疗脾阳虚型痰饮32例[J].上海针灸杂志,1996,15(3):55-56.
[23]陈仲杰,吴中朝,王巧妹,等.高脂血症痰饮病机及“温灸和之”治法探析[J].中医杂志,2012,53(17):1460-1461,1477.
[24]马泽云,曹毅.艾灸提高免疫功能的研究进展[J].浙江中医杂志,2004(3):133-135.
[25]葛建军,孙立虹,李新华,等.隔物灸治疗原发性痛经寒湿凝滞型的临床观察[J].北京中医药大学学报,2009,32(12):859-862.
[26]仲大奎.针灸对佐剂性关节炎大鼠TNF-α、IL-1、PGE_2及AQP1影响的实验研究[D].北京:北京中医药大学,2010.
[27]郑仕平,韩为,储浩然,等.通督调神针灸预处理对脑缺血再灌注大鼠相关微小RNA调控机制的研究[J].针刺研究,2015,40(2):99-103.
[28]张眉.托伐普坦治疗心力衰竭的系统评价和Meta分析[D].重庆:重庆医科大学,2014.
[29]Takeda T,Sawada S,Takeda S,et al.The effects of V2 antagonist(OPC-31260)on endolymphatic hydrops[J].Hear Res,2003,182(1-2):9-18.
[30]李晓泓,韩毳,张露芬,等.艾灸“大椎”穴抗应激作用的实验研究[J].中国行为医学科学,2002,11(5):495-497.
[31]Azzena G B,Melis F,Caria M A,et al.Vestibular projections to hypothalamic supraoptic and paraventricular nuclei[J].Arch Ital Biol,1993,131(2-3):127-136.
[32]许焕芳,赵百孝.艾灸疗法作用机理浅述[J].上海针灸杂志,2012,31(1):6-9.
[33]张荣伟.艾灸百会穴治疗内耳性眩晕66例[J].上海针灸杂志,2010,29(12):812.
[34]周敬华,刘传珍,庄伟华,等.精氨酸加压素对大鼠脑水肿形成及水孔蛋白-4表达的影响[J].临床神经病学杂志,2010,23(1):46-48.
[35]李琦,黄德亮.内耳水通道蛋白功能的研究进展[J].广东医学,2004,25(12):1476-1478.
[36]李学军.水通道的生物学研究[J].生命科学,2003,15(6):383-390.
[2]Hallpike C S,Cairns H.Observations on the Pathology of Meniere's Syndrome:(Section of Otology)[J].Proc R Soc Med,1938,31(11):1317-1336.
[3]张大帅,穆帅,刘颖,等.非肽类精氨酸加压素受体拮抗剂的研究进展[J].中国药物化学杂志,2014,24(2):103-115.
[4]Kakigi A,Takeda T.Antidiuretic hormone and osmolality in patients with Meniere's disease[J].ORL J Otorhinolaryngol Relat Spec,2009,71(1):11-13.
[5]Sawada S,Takeda T,Saito H.Antidiuretic hormone and psychosomatic aspects in Meniere's disease[J].Acta Otolaryngol Suppl,1997,528:109-112.
[6]Kitano H,Takeda T,Takeda S,et al.Endolymphatic hydrops by administration of vasopressin in the rat[J].Acta Histochem,Cytoc,2001,34(4):229-233.
[7]Gu F M,Han H L,Zhang L S.Effects of vasopressin on gene expression in rat inner ear[J].Hear Res,2006,222(1-2):70-78.
[8]Nishimoto G,Zelenina M,Li D,et al.Arginine vasopressin stimulates phosphorylation of aquaporin-2 in rat renal tissue[J].Am J Physiol,1999,276(2 Pt 2):F254-F259.
[9]Kumagami H,Loewenheim H,Beitz E,et al.The effect of anti-diuretic hormone on the endolymphatic sac of the inner ear[J].Pflugers Arch,1998,436(6):970-975.
[10]Long A F,Xing M,Morgan K,et al.Exploring the Evidence Base for Acupuncture in the Treatment of Ménière's Syndrome—A Systematic Review[J].Evidence-Based Complementary and Alternative Medicine,2011,2011:1-13.
[11]蒋丽元,倪芳英,王灿军,等.不同针灸方法对膜迷路积水模型豚鼠听力及耳蜗形态结构的影响[J].中国比较医学杂志,2015,25(2):22-25,33.
[12]Takeda T,Taguchi D.Aquaporins as potential drug targets for Meniere's disease and its related diseases[J].Handb Exp Pharmacol,2009(190):171-184.
[13]马海珠,钟时勋,温雅.醋酸去氨加压素对豚鼠耳蜗水通道蛋白2表达的调节作用[J].听力学及言语疾病杂志,2014,22(3):286-289.
[14]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:314-329.
[15]中国中医研究院针灸研究所.标准针灸经穴部位图谱[M].北京:中国标准出版社,1990:54,84.
[16]Naganuma H,Kawahara K,Tokumasu K,et al.Effects of arginine vasopressin on auditory brainstem response and cochlear morphology in rats[J].Auris Nasus Larynx,2014,41(3):249-254.
[17]黄菁.苓桂术甘汤合泽泻汤对豚鼠膜迷路积水模型小肠AQP8表达影响的研究[D].成都:成都中医药大学,2011.
[18]Takeda T,Takeda S,Kitano H,et al.Endolymphatic hydrops induced by chronic administration of vasopressin[J].Hear Res,2000,140(1-2):1-6.
[19]Takumida M,Kakigi A,Egami N,et al.Localization of aquaporins 1,2,and 3 and vasopressin type 2 receptor in the mouse inner ear[J].Acta Otolaryngol,2012,132(8):807-813.
[20]陈婷,张榕.水通道蛋白2在内淋巴囊上皮和肾脏中表达的研究[J].听力学及言语疾病杂志,2008,16(5):405-409.
[21]Sawada S,Takeda T,Kitano H,et al.Aquaporin-2 regulation by vasopressin in the rat inner ear[J].Neuroreport,2002,13(9):1127-1129.
[22]周道高.灸疗脾阳虚型痰饮32例[J].上海针灸杂志,1996,15(3):55-56.
[23]陈仲杰,吴中朝,王巧妹,等.高脂血症痰饮病机及“温灸和之”治法探析[J].中医杂志,2012,53(17):1460-1461,1477.
[24]马泽云,曹毅.艾灸提高免疫功能的研究进展[J].浙江中医杂志,2004(3):133-135.
[25]葛建军,孙立虹,李新华,等.隔物灸治疗原发性痛经寒湿凝滞型的临床观察[J].北京中医药大学学报,2009,32(12):859-862.
[26]仲大奎.针灸对佐剂性关节炎大鼠TNF-α、IL-1、PGE_2及AQP1影响的实验研究[D].北京:北京中医药大学,2010.
[27]郑仕平,韩为,储浩然,等.通督调神针灸预处理对脑缺血再灌注大鼠相关微小RNA调控机制的研究[J].针刺研究,2015,40(2):99-103.
[28]张眉.托伐普坦治疗心力衰竭的系统评价和Meta分析[D].重庆:重庆医科大学,2014.
[29]Takeda T,Sawada S,Takeda S,et al.The effects of V2 antagonist(OPC-31260)on endolymphatic hydrops[J].Hear Res,2003,182(1-2):9-18.
[30]李晓泓,韩毳,张露芬,等.艾灸“大椎”穴抗应激作用的实验研究[J].中国行为医学科学,2002,11(5):495-497.
[31]Azzena G B,Melis F,Caria M A,et al.Vestibular projections to hypothalamic supraoptic and paraventricular nuclei[J].Arch Ital Biol,1993,131(2-3):127-136.
[32]许焕芳,赵百孝.艾灸疗法作用机理浅述[J].上海针灸杂志,2012,31(1):6-9.
[33]张荣伟.艾灸百会穴治疗内耳性眩晕66例[J].上海针灸杂志,2010,29(12):812.
[34]周敬华,刘传珍,庄伟华,等.精氨酸加压素对大鼠脑水肿形成及水孔蛋白-4表达的影响[J].临床神经病学杂志,2010,23(1):46-48.
[35]李琦,黄德亮.内耳水通道蛋白功能的研究进展[J].广东医学,2004,25(12):1476-1478.
[36]李学军.水通道的生物学研究[J].生命科学,2003,15(6):383-390.