针刺对哮喘大鼠气道重建模型气道平滑肌细胞T型钙通道蛋白表达的影响

Influence of acupuncture on expression of T-type calcium channel protein in airway smooth muscle cell in airway remodeling rats with asthma

目的:观察针刺抗哮喘气道重建效应,并探讨气道平滑肌细胞T型钙通道在哮喘气道重建效应中的作用机制。方法:将32只大鼠随机分为正常组、模型组、针刺组、假针刺组,每组8只。后3组均造模:大鼠一次性腹腔注射卵蛋白10mg和氢氧化铝200mg的生理盐水混悬液1mL以及1mL灭活百日咳菌苗处于致敏状态14d。模型组第15天起连续14d每天超声雾化吸入1%卵蛋白诱发哮喘30min。针刺组大鼠造模后亦连续14d每天超声雾化吸入1%卵蛋白前针刺"大椎""风门""肺俞",每次30min,每2天1次。假针刺组与针刺组取穴及疗程相同,造模后每天超声雾化吸入1%卵蛋白前针刺穴位皮肤1mm,不留针。正常组不予干预处理。检测各组大鼠气道阻力评价呼吸功能,采用肺组织病理学检测评价气道重建情况,并运用免疫组化技术检测大鼠气道平滑肌T型钙通道3个亚单位Cav3.1、Cav3.2、Cav3.3蛋白表达情况。结果:①模型组气道阻力高于正常组与针刺组(均P<0.05),而针刺组气道阻力低于假针刺组(P<0.05)。②模型组气道壁厚度与气道基底膜周径比值(Awt/Pbm)高于正常组及针刺组(均P<0.05),而针刺组Awt/Pbm比值小于假针刺组(P<0.05)。模型组气道外径与气道内径比值(Po/Pi)高于正常组和针刺组(均P<0.05),针刺组Po/Pi比值小于假针刺组(P<0.05)。③模型组气道平滑肌细胞Cav3.1、Cav3.2平均光密度均高于正常组、针刺组(均P<0.05),模型组气道平滑肌细胞Cav3.3平均光密度高于正常组(P<0.05),低于假针刺组(P<0.05)。结论:针刺能抑制哮喘气道重建,降低气道阻力,抑制气道平滑肌增生,其机制可能与抑制气道平滑肌细胞T型钙通道蛋白,尤其是Cav3.1蛋白表达水平相关。

Objective To observe the anti-remodeling effect of acupuncture on asthma and to explore the mechanism of T-type calcium channel protein in airway smooth muscle cell in airway remodeling effect in asthma.Methods Thirty-two rats were randomly divided into a normal group,a model group,an acupuncture group and a sham acupuncture group,8 rats in each group.The rats in the latter three groups were sensitized for consecutive 14 days by single peritoneal injection of aqueous suspension 1 mL of 10 mg Ovalbumin（OVA）,200 mg Aluminum Hydroxide and Saline together with 1 mL inactivated Pertussis Vaccine.From the 15th day,asthma was induced for 30 minutes by ultrasonic atomizing inhalation of 1% OVA for consecutive 14 days in the model group.The acupuncture group was treated with acupuncture at ＂Dazhui＂（GV 14）,＂Fengmen＂（BL 12） and ＂Feishu＂（BL 13） for 30 minutes before the ultrasonic atomizing inhalation,once every two days for consecutive 14 days.The same acupoints selection and the course of treatment as the acupuncture group were produced in the sham acupuncture group and they were treated with acupuncture at 1mm acupoint skin without retaining needles.The normal group remained unhandled.The respiratory function and the airway remodeling were evaluated by airway resistance and pulmonary histopathology,respectively,and the T-type calcium channel protein expression of Ca v 3.1 、 Ca v 3.2 、 Ca v 3.3in airway smooth muscle cell were detected by immunohistochemistry technique.Results ① The airway resistance in the model group was higher than that in the normal group and in the acupuncture group（both P0.05）,and the airway resistance in the acupuncture group was lower than that in the sham acupuncture group（P0.05）.② The ratios of the airway wall thickness to the basement membrane perimeter（Awt / Pbm） and the airway outer perimenter to the airway internal perimeter（Po / Pi） in the model group were higher than those in the normal group and in the acupuncture group（all P0.05）,and the ratios of Awt / Pbm and Po / Pi in the acupuncture group were lower than those in the sham acupuncture group（both P0.05）.③ The average optical of Ca v 3.1and Ca v 3.2in airway smooth muscle cell in the model group were higher than that in the normal group and in the acupuncture group（both P0.05）,and the average optical of Ca v 3.3in airway smooth muscle cell in the model group was higher than that in the normal group（P0.05） and it was lower than that in the sham acupuncture group（P0.05）.Conclusion Acupuncture can inhibit the airway remodeling and the accrementition of the airway smooth muscle and can reduce the airway resistance.The mechanism may be related to the inhibition of T-type calcium channel protein in airway smooth muscle cell,especially in relation to the protein expression of Cav3.1.