基于JAK2/STAT3信号通路探究利咽糖浆对慢性咽炎大鼠咽喉组织损伤及咽黏膜修复的影响

Influences of Liyan syrup on pharyngeal tissue injury and pharyngeal mucosal repair in rats with chronic pharyngitis based on JAK2/STAT3 signaling pathway

目的:基于Janus激酶2(JAK2)/信号转导和转录激活因子3(STAT3)信号通路探究利咽糖浆对慢性咽炎大鼠咽喉组织损伤及咽黏膜修复的影响。方法:取SD大鼠随机分为对照组、模型组、利咽糖浆组、RO8191(JAK2/STAT3激活剂)组、利咽糖浆+RO8191组,模型组与药物干预组大鼠采用氨水刺激咽部构建慢性咽炎模型,对照组大鼠咽部注射等剂量生理盐水,经利咽糖浆与RO8191干预后,检测大鼠一般情况及咽部表观状态,并进行咽部表观状态评分;HE染色检测大鼠咽部病理形态变化;流式细胞术检测大鼠外周血T淋巴细胞亚群CD4^(+)T与CD8^(+)T表达、CD4^(+)T/CD8^(+)T;试剂盒检测大鼠血清肿瘤坏死因子-α(TNF-α)、IL-6、IL-10、丙二醛(MDA)、活性氧(ROS)、总抗氧化能力(T-AOC)水平;以免疫印记法检测大鼠咽部组织JAK2/STAT3通路相关蛋白表达。结果:与对照组比较,模型组大鼠咽部组织出现明显病理形态损伤,外周血CD4^(+)T与CD4^(+)T/CD8^(+)T、IL-10、血清T-AOC水平降低(P<0.05),咽部表观状态评分、CD8^(+)T、血清TNF-α、IL-6、MDA与ROS水平、咽部组织p-JAK2/JAK2与p-STAT3/STAT3水平升高(P<0.05);与模型组、利咽糖浆+RO8191组相比,利咽糖浆组大鼠咽部组织病理形态损伤减轻,外周血CD4^(+)T表达与CD4^(+)T/CD8^(+)T、IL-10、血清T-AOC水平均升高(P<0.05),咽部表观状态评分、CD8^(+)T、血清TNF-α、IL-6、MDA与ROS水平、咽部组织p-JAK2/JAK2与p-STAT3/STAT3水平均降低(P<0.05);RO8191组大鼠各指标变化趋势与利咽糖浆组相反。结论:利咽糖浆可通过抑制JAK2/STAT3信号激活减轻炎症及氧化应激,增强抗炎及抗氧化能力,缓解慢性咽炎大鼠咽喉组织损伤,修复咽黏膜。

Objective:Based on the Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway,to explore the influences of Liyan syrup on pharyngeal tissue injury and pharyngeal mucosal repair in rats with chronic pharyngitis.Methods:SD rats were randomly separated into control group,model group,Liyan syrup group,RO8191(JAK2/STAT3 activator)group,Liyan syrup+RO8191 group,the model group and drug intervention group were treated with ammonia water to stimulate the pharynx to build a chronic pharyngitis model,while rats in control group were injected with an equal dose of normal saline into the pharynx.After the intervention of Liyan Syrup and RO8191,the general condition of rats and the apparent state of pharynx were detected,and the apparent state of pharynx was scored;the pathological changes of pharynx of rats were measured by HE staining;the proportions of CD4^(+)T and CD8^(+)T and the ratio of CD4^(+)T/CD8^(+)T in peripheral blood T lymphocyte subsets of rats were measured by flow cytometry;the levels of serum tumor necrosis factor alpha(TNF-α),IL-6,IL-10,malondialdehyde(MDA),reactive oxygen species(ROS)and total antioxidant capacity(T-AOC)were measured by kits;the expressions of JAK2/STAT3 pathway-related proteins in rat pharyngeal tissues were measured by immunoblotting.Results:Compared with control group,the pharyngeal tissue of model group showed obvious pathological morphological damage,the peripheral blood CD4^(+)T proportion and CD4^(+)T/CD8^(+)T,IL-10 and serum TAOC level decreased(P<0.05),the pharyngeal appearance state score,CD8^(+)T ratio,serum TNF-α,IL-6,MDA and ROS levels,and pharyngeal tissue p-JAK2/JAK2 and p-STAT3/STAT3 levels increased(P<0.05);compared with model group and Liyan syrup+RO8191 group,the pathological and morphological damage of the pharynx of rats in Liyan syrup group was alleviated,the peripheral blood CD4^(+)T and CD4^(+)T/CD8^(+)T,IL-10,and serum T-AOC level all increased(P<0.05),and the pharyngeal appearance state score,CD8^(+)T,serum TNF-α,IL-6,MDA and ROS levels,and pharyngeal tissue p-JAK2/JAK2 and p-STAT3/STAT3 levels all decreased(P<0.05);the change trend of each index in RO8191 group was opposite to that in Liyan syrup group.Conclusion:Liyan syrup can reduce inflammation and oxidative stress by inhibiting the activation of JAK2/STAT3 signal,enhance anti-inflammatory and antioxidant capacity,relieve throat tissue damage and repair pharyngeal mucosa in rats with chronic pharyngitis.