金嗓散结胶囊对化学刺激法致慢性喉炎大鼠的作用及机制研究

Effect and mechanism of Jinsang Sanjie Capsule on chronic laryngitis caused by chemical stimulation method

目的观察金嗓散结胶囊对化学刺激法所致的大鼠慢性喉炎模型的影响。方法使用10%的吡啶溶液对大鼠喉部进行刺激,每周2次,共3周,造模结束后,将造模50只大鼠随机分为5组:模型组、清咽利喉颗粒(阳性药,1.58 g·kg^(-1))组和金嗓散结胶囊高、中、低剂量(0.67、0.33、0.17 g·kg^(-1))组。ig给药,每天给药1次,共4周,对照组和模型组每天ig等体积的纯净水。给药过程中观察各组大鼠一般状态;ELISA法检测血清白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、细胞间黏附分子-1(ICAM-1)水平;HE染色后进行喉部组织病理学检查;Western blotting法检测喉部声带组织TNF-α、C-反应蛋白(CRP)、基质金属蛋白酶-9(MMP-9)、基质金属蛋白酶抑制剂-1(TIMP1)、IL-6蛋白表达水平。结果与模型组比较,金嗓散结胶囊可以减轻大鼠化学刺激法所致慢性喉炎的声音沙哑等症状,减轻喉部黏膜增生和坏死、炎细胞浸润等病理症状;金嗓散结胶囊高、中剂量组血清IL-6、TNF-α、IL-1β、ICAM-1水平显著降低(P<0.05、0.01),低剂量组血清TNF-α水平显著降低(P<0.05);高、中、低剂量组声带组织TNF-α、CRP,高、中剂量组MMP-9、IL-6以及高剂量组TIMP1蛋白相对表达水平均显著降低(P<0.05、0.01)。结论金嗓散结胶囊可以减轻化学刺激法所致大鼠慢性喉炎,其作用机制可能与下调促炎因子TNF-α、IL-6进而降低ICAM-1的表达,抑制CRP进而降低溶蛋白酶造成的有害作用,降低MMP-9蛋白表达进而抑制基底膜增厚有关。

Objective To observe the effect of Jinsang Sanjie Capsule on the model of chronic laryngitis in rats caused by chemical stimulation.Methods The larynx of rats was stimulated by 10%pyridine solution twice a week for a total of three weeks,and after the molding was completed,50 SD rats in the module were divided into five groups,namely model group,Qingyan Lihou Granules(positive drug,1.58 g·kg^(-1))group and Jinsang Sanjie Capsule high,medium and low-dose(0.67,0.33,and 0.17 g·kg^(-1))groups.The drug was ig given once a day for four weeks,and the control group and model group were given equal volume of pure water ig every day.Serum levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and intercellular adhesion molecule-1(ICAM-1)were determined by Elisa.Laryngeal histopathology was performed after HE staining.The expression levels of TNF-α,Creactive protein(CRP),matrix metalloproteinase-9(MMP-9),matrix metalloproteinase-inhibitor-1(TIMP1),and IL-6 protein in larynx were detected by Western blotting.Results Compared with model group,Jinsang Sanjie Capsule could alleviate the hoarseness and other symptoms of chronic laryngitis caused by chemical stimulation in rats,and alleviate the pathological symptoms of laryngeal mucosa hyperplasia and necrosis,inflammatory cell infiltration and so on.The serum levels of IL-6,TNF-α,IL-1βand ICAM-1 in Jinsang Sanjie Capsule high and medium dose groups were significantly decreased compared with model group(P<0.01,0.05),and the serum levels of TNF-αin low dose group were significantly decreased(P<0.05).The relative expression levels of TNF-αand CRP in vocal cord tissue in high,medium and low dose groups,MMP-9 and IL-6 in high and medium dose groups and TIMP1 in high dose groups were significantly decreased compared with model group(P<0.01,0.05).Conclusion Jinsang Sanjie capsule can alleviate chronic laryngitis induced by chemical stimulation in rats,and its mechanism may be related to the down-regulation of pro-inflammatory factors TNF-αand IL-6,thus reducing the expression of ICAM-1,inhibiting CRP,thus reducing the harmful effect caused by lysoprotease,and reducing the expression of MMP-9 protein,thus inhibiting the thickening of basement membrane.