目的探讨JNK信号通路抑制剂(SP600125)体内长期实验的应用方法。方法将40只新西兰兔随机分为八组:空白对照组、模型对照组、JNK高剂量组、JNK低剂量组、JNK空白组、DMSO模型组、DMSO空白组、阳性对照组。自实验第一天开始各组给予相应处...目的探讨JNK信号通路抑制剂(SP600125)体内长期实验的应用方法。方法将40只新西兰兔随机分为八组:空白对照组、模型对照组、JNK高剂量组、JNK低剂量组、JNK空白组、DMSO模型组、DMSO空白组、阳性对照组。自实验第一天开始各组给予相应处理,连续4周。4周末检测实验兔心肌组织JNK总蛋白及蛋白磷酸化和基因表达水平。结果经阿霉素干预后,各组实验组心肌P-JNK表达均不同程度上调,其中以模型对照组、DMSO模型组上调最为明显(P<0.05)。JNK低、高剂量组和阳性对照组心肌P-JNK表达水平上调较模型对照组缓慢(P<0.05),其中以JNK高剂组心肌P-JNK水平上调最为缓慢(P<0.05)。各实验组间JNK总蛋白、JNK m RNA表达差异无统计学意义(P>0.05)。结论连续4周小剂量皮下注射JNK信号通路抑制剂可以显著下调p-JNK的表达,该方法能够为体内长期实验研究提供思路。展开更多
Background: We wanted to determine (via an immunopharmacological approach) whether the c-Jun NH2 terminal kinase (JNK) cascade is phosphorylated in the submandibular gland by carbachol and cholinergic autoantibodies (...Background: We wanted to determine (via an immunopharmacological approach) whether the c-Jun NH2 terminal kinase (JNK) cascade is phosphorylated in the submandibular gland by carbachol and cholinergic autoantibodies (IgG) present in the sera of patients with primary Sj?gren’s syndrome (pSS) by interaction and activation of salivary gland muscarinic acetylcholine receptors (mAChRs). Methods: The JNK, PGE2 and NOS assays were measured in rat sub- mandibular gland with pSS IgG and carbachol alone or in the presence of different blocker agents. Results: pSS IgG- activated M3 mAChRs stimulated JNK phosphorylation whereas the activation of M1 mAChRs by carbachol stimulated JNK phosphorylation involving calcium-activated mechanism. The intracellular pathway leading to pSS IgG-induced biological effects on JNK activity involved activation of protein kinase C (PKC), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) enzymes. Also, activation of COX-2 and COX-1 by pSS IgG and carbachol-induced PGE2 generation were involved. Conclusion: These results may contribute to better understanding the modulatory role of JNK enzymes by cholinergic autoantibodies from pSS patients acting on mAChR in rat submandibular gland.展开更多
文摘目的探讨JNK信号通路抑制剂(SP600125)体内长期实验的应用方法。方法将40只新西兰兔随机分为八组:空白对照组、模型对照组、JNK高剂量组、JNK低剂量组、JNK空白组、DMSO模型组、DMSO空白组、阳性对照组。自实验第一天开始各组给予相应处理,连续4周。4周末检测实验兔心肌组织JNK总蛋白及蛋白磷酸化和基因表达水平。结果经阿霉素干预后,各组实验组心肌P-JNK表达均不同程度上调,其中以模型对照组、DMSO模型组上调最为明显(P<0.05)。JNK低、高剂量组和阳性对照组心肌P-JNK表达水平上调较模型对照组缓慢(P<0.05),其中以JNK高剂组心肌P-JNK水平上调最为缓慢(P<0.05)。各实验组间JNK总蛋白、JNK m RNA表达差异无统计学意义(P>0.05)。结论连续4周小剂量皮下注射JNK信号通路抑制剂可以显著下调p-JNK的表达,该方法能够为体内长期实验研究提供思路。
文摘Background: We wanted to determine (via an immunopharmacological approach) whether the c-Jun NH2 terminal kinase (JNK) cascade is phosphorylated in the submandibular gland by carbachol and cholinergic autoantibodies (IgG) present in the sera of patients with primary Sj?gren’s syndrome (pSS) by interaction and activation of salivary gland muscarinic acetylcholine receptors (mAChRs). Methods: The JNK, PGE2 and NOS assays were measured in rat sub- mandibular gland with pSS IgG and carbachol alone or in the presence of different blocker agents. Results: pSS IgG- activated M3 mAChRs stimulated JNK phosphorylation whereas the activation of M1 mAChRs by carbachol stimulated JNK phosphorylation involving calcium-activated mechanism. The intracellular pathway leading to pSS IgG-induced biological effects on JNK activity involved activation of protein kinase C (PKC), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) enzymes. Also, activation of COX-2 and COX-1 by pSS IgG and carbachol-induced PGE2 generation were involved. Conclusion: These results may contribute to better understanding the modulatory role of JNK enzymes by cholinergic autoantibodies from pSS patients acting on mAChR in rat submandibular gland.
