摘要
目的:探讨桑杏止咳方对咳嗽变异性哮喘(cough variant asthma,CVA)细胞焦亡的影响。方法:将36只SD雄性大鼠随机分为6组:空白对照组、模型组、地塞米松组及桑杏止咳方低、中、高剂量组,每组6只。采用卵蛋白(ovalbumin,OVA)致敏与雾化激发法制备CVA模型。各给药组于造模第18天开始灌胃相应药物,连续14 d。检测各组大鼠气道阻力;采用苏木素-伊红(hematoxylin-eosin,HE)染色观察肺组织病理变化;通过酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)法检测血清核苷酸结合寡聚化结构域样受体蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)、胱天蛋白酶-1(cysteinyl aspartate specific proteinase-1,Caspase-1)、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-18(interleukin-18,IL-18)等因子水平;采用蛋白免疫印迹(Western blot)法检测肺组织NLRP3、Caspase-1、消皮素D(gasdermin D,GSDMD-N)蛋白表达;通过实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)法检测肺组织NLRP3、Caspase-1、GSDMD-N、IL-1β、IL-18的mRNA表达水平。结果:与空白对照组对比,模型组大鼠气道阻力显著升高(P<0.05),肺组织炎症细胞浸润加重,血清NLRP3、Caspase-1、IL-1β、IL-18水平及肺组织Cleaved Caspase-1、NLRP3、GSDMD-N蛋白表达均显著上调(均P<0.05),肺组织NLRP3、Caspase-1、GSDMD-N、IL-1β、IL-18的mRNA表达水平均显著升高(均P<0.05)。与模型组对比,桑杏止咳方各剂量组气道阻力显著降低(P<0.05),肺组织病理损伤减轻,血清焦亡相关因子水平及肺组织焦亡关键蛋白与mRNA表达水平均显著下调(均P<0.05),且高剂量组效果最为显著。结论:桑杏止咳方可有效改善CVA模型大鼠的气道高反应性及肺组织炎症损伤,其作用机制可能与抑制NLRP3/Caspase-1信号通路,进而减轻细胞焦亡有关。
Objective:To investigate the effects of Sangxing Zhike Formula on pyroptosis in cough variant asthma(CVA).Methods:Thirty six male SD rats were randomly divided into six groups:blank control group,model group,dexamethasone group,and low,medium,and high dose Sangxing Zhike Formula groups,with six rats in each group.The CVA model was established using ovalbumin(OVA)sensitization combined with aerosol stimulation.The drug administration groups received the respective treatments by gavage starting on day 18 after modeling,for 14 consecutive days.Airway resistance was measured in each group;lung tissue pathological changes were observed using hematoxylin eosin(HE)staining;serum levels of NOD-like receptor thermal protein domain associated protein 3(NL-RP3),cysteinyl aspartate specific proteinase−1(Caspase−1),interleukin−1β(IL-1β),and interleukin-18(IL-18)were determined using enzyme linked immunosorbent assay(ELISA);protein expression of NLRP3,Caspase-1,and gasdermin D(GSDMD N)in lung tissue was detected by Western blot;and mRNA expression levels of NLRP3,Caspase-1,GSDMD-N,IL 1β,and IL 18 in lung tissue were measured by quantitative real time PCR(qRT PCR).Results:Compared with the blank control group,airway resistance in model rats was significantly increased(P<0.05),lung tissue showed aggravated inflammatory cell infiltration,serum levels of NLRP3,Caspase-1,IL-1β,and IL 18,as well as lung tissue expression of Cleaved Caspase-1,NLRP3,and GSDMD-N N proteins,were all significantly upregulated(all P<0.05),and mRNA expression levels of NLRP3,Caspase−1,GSDMD N,IL 1β,and IL 18 in lung tissue were significantly elevated(all P<0.05).Compared with the model group,all doses of Sangxing Zhike Formula significantly reduced airway resistance(P<0.05),alleviated lung tissue pathological damage,and significantly downregulated serum levels of pyroptosis related factors,as well as key proteins and mRNA expression in lung tissue(all P<0.05),with the high dose group showing the most pronounced effect.Conclusion:Sangxing Zhike Formula can effectively improve airway hyperresponsiveness and lung tissue inflammatory damage in CVA model rats,and its mechanism may be related to inhibiting the NLRP3/Caspase-1 signaling pathway,thereby reducing cell pyroptosis.
作者
赵永伟
张晓谊
罗建江
ZHAO Yongwei;ZHANG Xiaoyi;LUO Jianjiang(Urumqi City Hospital of Traditional Chinese Medicine,Urumqi Xinjiang China 830000;Traditional Chinese Medicine Hospital Affiliated to Xinjiang Medical University,Urumqi Xinjiang China 830000)
出处
《中医学报》
2026年第1期176-182,共7页
Acta Chinese Medicine
基金
国家自然科学基金项目(82060794)
杰出青年科学基金项目(2022D01E80)
新疆维吾尔自治区自然科学基金面上项目(2022D01A33)。
