基于JAK2/STAT3信号通路探讨升清降浊制动颗粒对多发性抽动症模型大鼠的作用及机制

Exploring the effect and mechanism of Shengqing Jiangzhuo Zhidong Granule on Tourette’s syndrome model rats based on JAK2/STAT3 signal pathway

[目的]基于Janus激酶2(JAK2)/信号转导和转录激活子3(STAT3)信号通路探讨升清降浊制动颗粒对多发性抽动症(TS)模型大鼠的作用及机制。[方法]将SD大鼠分为对照组、TS组、升清降浊制动颗粒低剂量组(0.645 mg/kg)、升清降浊制动颗粒高剂量组(2.58 mg/kg)、氟哌啶醇组(200μg/kg)、Coumermycin A1(JAK2通路激活剂)组(4 mg/kg)、升清降浊制动颗粒高剂量+Coumermycin A1组(2.58 mg/kg+4 mg/kg),每组12只。除对照组外,其他组大鼠均需构建TS模型。建模成功后,进行给药处理,给药每日1次,持续21 d。检测大鼠运动行为评分、刻板行为评分变化;酶联免疫吸附实验(ELISA)检测血清中白细胞介素(IL)-1β、肿瘤坏死因子-α(TNF-α)、IL-6水平及纹状体中多巴胺(DA)、多巴胺转运蛋白(DAT)、多巴胺D2受体(DRD2)含量;DNA缺口末端标记法(TUNEL)染色检测大鼠纹状体中神经元凋亡;蛋白免疫印迹法(Western Blot)检测大鼠纹状体中p-JAK2、p-STAT3蛋白表达。[结果]与对照组比较,TS组大鼠运动行为评分、刻板行为评分、IL-1β、TNF-α、IL-6水平、DRD2含量、神经元凋亡率、p-JAK2、pSTAT3蛋白表达升高,DA、DAT含量降低(P<0.05)。与TS组比较,升清降浊制动颗粒低剂量组、升清降浊制动颗粒高剂量组、氟哌啶醇组大鼠运动行为评分、刻板行为评分、IL-1β、TNF-α、IL-6水平、DRD2含量、神经元凋亡率、pJAK2、p-STAT3蛋白表达降低,DA、DAT含量升高(P<0.05)。与TS组比较,Coumermycin A1组大鼠运动行为评分、刻板行为评分、IL-1β、TNF-α、IL-6水平、DRD2含量、神经元凋亡率、p-JAK2、p-STAT3蛋白表达升高,DA、DAT含量降低(P<0.05)。Coumermycin A1减弱了高剂量升清降浊制动颗粒对TS模型大鼠的保护作用。[结论]升清降浊制动颗粒可抑制TS大鼠神经炎症、神经元凋亡及促进DA平衡,该机制可能与抑制JAK2/STAT3信号通路有关。

[Objective]To explore the effect and mechanism of Shengqing Jiangzhuo Zhidong Granule on Tourette’s syndrome(TS)model rats based on Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signal pathway.[Methods]SD rats were grouped into control group,TS group,low dose Shengqing Jiangzhuo Zhidong Granule group(0.645 mg/kg),high dose Shengqing Jiangzhuo Zhidong Granule group(2.58 mg/kg),haloperidol group(200μg/kg),Coumermycin A1(JAK2 pathway activator)group(4 mg/kg),and high dose Shengqing Jiangzhuo Zhidong Granule+Coumermycin A1 group(2.58 mg/kg+4 mg/kg),12 in each group.Except for the control group,rats in other groups were all constructed with TS model.After successful modeling,the drug was administered once a day for 21 days.The changes in motor behavior score and stereotyped behavior score of rats were detected;the levels of serum interleukin(IL)-1β,tumor necrosis factor-α(TNF-α),IL-6,and the contents of dopamine(DA),dopamine transporter(DAT)and dopamine D2 receptor(DRD2)in striatum were detected by enzyme-linked immunosorbent assay(ELISA);DNA notch end labeling(TUNEL)staining was applied to detect neuronal apoptosis in rat striatum;Western blot was applied to detect the expression of p-JAK2 and p-STAT3 proteins in rat striatum.[Results]Compared with the control group,the motor behavior score,stereotyped behavior score,IL-1β,TNF-α,IL-6 levels,DRD2 content,neuron apoptosis rate,p-JAK2,p-STAT3 protein expression in TS group were higher,and the DA and DAT contents were lower(P<0.05);compared with TS group,the motor behavior score,stereotyped behavior score,IL-1β,TNF-α,IL-6 levels,DRD2 content,neuron apoptosis rate,p-JAK2,p-STAT3 protein expression of rats in the low dose Shengqing Jiangzhuo Zhidong Granule group,high dose Shengqing Jiangzhuo Zhidong Granule group and haloperidol group were lower,and the DA and DAT contents were higher(P<0.05);compared with TS group,the scores of motor behavior,stereotyped behavior,IL-1β,TNF-α,IL-6 levels,DRD2 content,neuron apoptosis rate,p-JAK2,p-STAT3 protein expression of rats in Coumermycin A1 group were higher,and the DA and DAT contents were lower(P<0.05);coumermycin A1 attenuated the protective effect of high-dose Shengqing Jiangzhuo Zhidong Granules on TS model rats.[Conclusion]Shengqing Jiangzhuo Zhidong Granules can inhibit neuroinflammation,neuron apoptosis and promote DA balance in TS rats,which may be related to the inhibition of JAK2/STAT3 signaling pathway.