半夏芩连汤对干燥综合征模型NOD小鼠Th17/IL-17免疫炎性途径的影响

Effect of Banxia Qinlian Decoction on Th17 / IL-17 Immune Inflammatory Way of Sjgren's Syndrome NOD Model Mice

目的探讨干燥综合征(Sjgren's syndrome,SS)外分泌腺免疫炎性损伤的分子机制及半夏芩连汤的干预作用。方法将18只雌性NOD小鼠随机分为模型组、阳性药组、中药组,每组6只,将6只雌性Balb/c小鼠设为空白对照组。空白对照组及模型组每天予去离子水0.1 m L/10 g灌胃,阳性药组予雷公藤多甙片10mg/kg灌胃,中药组每天予半夏芩连汤按生药60 g/kg灌胃。连续给药12周后,摘眼球取血处死,留取双侧腮腺、颌下腺组织。采用RT-PCR技术检测IL-17、IL-6、毒蕈碱样胆碱能受体3(type 3 muscarinic acetylcholine receptors,M3R)、水通道蛋白5(aquaporin protein-5,AQP5)mRNA转录水平,Western blot技术检测M3R、AQP5蛋白表达水平,ELISA法检测IL-17、IL-6蛋白表达水平。结果与正常组比较,模型组IL-17、IL-6、M3R、AQP5mRNA转录及蛋白的表达水平显著上调,差异有统计学意义(P<0.01)。与模型组比较,阳性药组、中药组IL-17、IL-6、M3R、AQP5 mRNA转录水平及IL-17、IL-6、M3R、AQP5蛋白表达显著下调,差异有统计学意义(P<0.01,P<0.05)。治疗组间比较,与中药组比较,阳性药组IL-17、IL-6、M3R mRNA转录水平显著下调(P<0.01),M3R、AQP5蛋白表达显著下调(P<0.01)。结论半夏芩连汤抑制SS外分泌腺神经毒性损伤的分子机制可能与其调控Th 17/IL-17免疫炎性途径相关。

Objective To explore the molecular mechanism of exocrine immune inflammatory injury of Sjsgren＇s Syndrome and the intervention of Banxia Qinlian Decoction （BQD）. Methods Totally 18 female NOD mice were randomly divided into the model group, the positive drug group, and the BQD group, 6 in each group. Six female BALB/c mice were recruited as a blank control group. Mice in the blank control group and the model group were gavaged with deionized water at the daily dose of 0.1 mL/10 g body weight. Tripterygium Tablet was administered by gastrogavage to mice in the positive group at the daily dose of 10 mg/kg. BQD was administered by gastrogavage to mice in the BQD group at the daily dose of 60 g crude drugs/kg. After 12 weeks of medication, mice were sacrificed. Their eyeballs were ex- cised and blood collected. Tissues of bilateral parotids and submandibular glands were kept. mRNA tran- scriptional levels of IL-17, IL-6, type 3 muscarinic acetylcholine receptors （M3R）, aquaporin protein-5 （AQP5） were detected by RT-PCR. Expression levels of M3R and AQP5 protein were detected by Western blot. Protein expression levels of IL-17 and IL-6 were detected by ELISA. Results Compared with the normal group, mRNA transcriptional levels and protein expression levels of IL-17, IL-6, M3R, and AQP5 were significantly up-regulated in the model group （P 〈0.01 ）. Compared with the model group, mRNA transcriptional levels and protein expression levels of IL-17, IL-6, M3R, and AQP5 were significantly down-regulated in the positive drug group and the BQD group with statistical difference （P 〈0.01, P 〈 0.05）. Compared with the BQD group, mRNA transcriptional levels of IL-17, IL-6, and M3R, as well as M3R and AQP5 protein expression levels were significantly down-regulated in the positive drug group （all P 〈 0.01 ）. Conclusion The molecular mechanism of BQD in inhibiting SS exocrine neurotoxic injury might be possibly related to regulating Th17/IL-17 immune inflammatory way.