射干麻黄汤及其辛、苦味药组拆方对寒哮大鼠气道炎症及肺组织TRPV1/TAS2R14表达的影响  

Effect of Shegan Mahuangtang and Its Pungent and Bitter Chinese Herbs on Airway Inflammation and Expression of TRPV1/TAS2R14 in Lung Tissue of Rat Model of Cold Asthma

作　　者：袁亚美[1,2] 叶卫东 程悦 李秋慧 刘家鑫[1] 乔佳乐 王坤 方向明[1,2,4] YUAN Yamei;YE Weidong;CHENG Yue;LI Qiuhui;LIU Jiaxin;QIAO Jiale;WANG Kun;FANG Xiangming(Anhui University of Chinese Medicine,Hefei 230012,China;Anhui Province Key Laboratory of Application and Transformation of Traditional Chinese Medicine in Prevention and Treatment of Major Pulmonary Diseases,Hefei 230031,China;The Second Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China;Key Laboratory of Xin'an Medicine,Ministry of Education,Hefei 230038,China)

机构地区：[1]安徽中医药大学,合肥230012 [2]中医药防治肺系重大疾病应用转化安徽省重点实验室,合肥230031 [3]安徽中医药大学第二附属医院,合肥230031 [4]新安医学教育部重点实验室,合肥230038

出　　处：《中国实验方剂学杂志》2024年第21期1-9,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(82074401);国家自然科学基金青年基金项目(82305187);安徽省教育厅自然科学研究重点项目(2022AH050446,2023AH050821);安徽中医药大学自然科学研究重点项目(2021zrzd18);中医药防治肺系重大疾病应用转化安徽省重点实验室开放课题(2024ZYFBAHKLA09)。

摘　　要：目的:探讨射干麻黄汤辛、苦味药组对哮喘寒哮证大鼠肺组织中瞬时受体电位通道香草醛亚型1(TRPV1)、苦味受体14(TAS2R14)蛋白表达的影响。方法:70只SD大鼠随机分为正常组、模型组、射干麻黄汤组、辛味药组、苦味药组、地塞米松组、桂龙咳喘宁组,10%卵清蛋白(OVA)结合氢氧化铝腹腔及四肢皮下注射致敏,联合2%OVA雾化及寒冷(2~4℃)刺激,激发大鼠寒哮模型。射干麻黄汤、辛味药、苦味药(6.43 g·kg^(-1))、地塞米松(0.5 g·kg^(-1))、桂龙咳喘宁组(10 g·kg^(-1))灌胃及雾化,空白组、模型组以等量生理盐水灌胃及雾化,持续干预3周。末次激发后,对大鼠肺组织进行苏木素-伊红(HE)、碘酸雪夫(PAS)、马松(Masson)染色观察气道炎症反应和细胞增殖情况。采用酶联免疫吸附测定法(ELISA)检测血清中白细胞介素-5(IL-5)、肿瘤坏死因子-α(TNF-α)、胸腺基质淋巴细胞生成素(TSLP)、转化生长因子-β_(1)(TGF-β_(1))含量;免疫荧光法检测TRPV1、TAS2R14的表达;蛋白免疫印迹法(Western blot)检测大鼠肺组织TRPV1、TAS2R14、磷脂酶Cβ(2 PLCβ_(2))、B细胞淋巴瘤-2(Bcl-2)、α-平滑肌肌动蛋白(α-SMA)表达。结果:与空白组比较,模型组大鼠饮水、饮食及体质量下降,气道炎性细胞浸润、杯状细胞增生、组织纤维化及胶原蛋白沉积情况增多,血清IL-5、TNF-α、TSLP和TGF-β_(1)含量升高(P<0.01),TRPV1、PLCβ_(2)、α-SMA的表达增加、TAS2R14、Bcl-2的表达降低(P<0.05,P<0.01);与模型组比较,射干麻黄汤、辛味药及苦味药物组大鼠饮水、饮食及体质量改善,气道炎性细胞浸润、杯状细胞增生、组织纤维化改变及胶原蛋白沉积情况减少;血清中IL-5、TNF-α、TSLP和TGF-β_(1)含量降低(P<0.01),TRPV1、PLCβ_(2)、α-SMA的表达减少,TAS2R14、Bcl-2表达增加(P<0.05,P<0.01)。结论:射干麻黄汤及其辛、苦味药物组可以明显减轻OVA诱发的哮喘大鼠气道炎症,降低TRPV1、PLCβObjective:To investigate the effects of Shegan Mahuangtang and its pungent and bitter Chinese herbs on the expression of transient receptor potential vanilloid-1(TRPV1)and bitter taste receptor 14(TAS2R14)in the lung tissue of the rat model of cold asthma.Method:Seventy SD rats were randomized into 7 groups:normal,model,Shegan Mahuangtang,pungent Chinese herbs,bitter Chinese herbs(6.43 g·kg^(-1)),dexamethasone(0.5 g·kg^(-1)),and Guilong Kechuanning(10 g·kg^(-1)).The rat model of cold asthma was established by intraperitoneal injection and subcutaneous injection of 10%ovalbumin(OVA)and aluminium hydroxide in the limbs,combined with 2%OVA atomization and cold(2-4℃)stimulation.The rats were treated with corresponding drugs by gavage and atomization,and the normal and model groups were treated with the same amount of normal saline for 3 weeks.After the last excitation,airway inflammation and cell proliferation were observed by hematoxylin-eosin(HE),periodic acid-Schiff(PAS),and Masson staining of the lung tissue.The levels of interleukin-5(IL-5),tumor necrosis factor-α(TNF-α),thymic stromal lymphopoietin(TSLP),and transforming growth factor-β_(1)(TGF-β_(1))in the serum were measured by enzymelinked immunosorbent assay(ELISA).The expression of TRPV1 and TAS2R14 was detected by immunofluorescence.The expression of TRPV1,TAS2R14,phospholipase Cβ_(2)(PLCβ_(2)),B-cell lymphoma-2(Bcl-2),andα-smooth muscle actin(α-SMA)in the lung tissue was determined by Western blot.Result:Compared with the normal group,the model group showed decreased water intake,food intake,and body weight,increased airway inflammatory cell infiltration,goblet cell proliferation,tissue fibrosis and collagen deposition,elevated levels of IL-5,TNF-α,TSLP,and TGF-β_(1)in the serum(P<0.01),upregulated expression of TRPV1,PLCβ_(2),andα-SMA,and downregulated expression of TAS2R14 and Bcl-2(P<0.05,P<0.01).Compared with model group,Shecgan Mahuangtang,pungent Chinese herbs,and bitter Chinese herbs increased the water intake,food intake,and body

关 键 词：辛味药 苦味药 寒哮 瞬时受体电位通道香草醛亚型1(TRPV1) 苦味受体14(TAS2R14) 

分 类 号：R2-0[医药卫生—中医学] R33R289R318.13