4,4′-二异硫氰基芪-2,2′-二磺酸对脓毒症小鼠心肌炎性反应的作用及机制
[摘要] 目的 探討4，4′-二异硫氰基芪-2，2′-二磺酸(DIDS)对脓毒症小鼠心肌炎性反应的作用及机制。 方法 雄性BALB/c小鼠总计52只，采用随机数字表法分为4组，分别为空白对照组、脂多糖(LPS)组、DIDS低剂量组和DIDS高剂量，每组13只。预处理3 d，其中DIDS低剂量组、DIDS高剂量组每天分别腹腔注射7、14 mg/kg的DIDS，空白对照组、LPS组每天腹腔注射等量的磷酸缓冲盐溶液(PBS)。第3天预处理完成2 h后，空白对照组腹腔注射PBS，其余三组腹腔注射LPS 10 mg/kg建立小鼠脓毒症模型。在模型创建成功后，每组10只小鼠进行48 h生存状况观察，计算存活率。另外每组3只小鼠于4 h后处死，取心脏组织，采用real-time PCR法检测炎性因子[包括肿瘤坏死因子-α(TNF-α)、白细胞介素1β(IL-1β)和白细胞介素-6(IL-6)]及Notch信号通路相关基因(包括Notch1～4、Dll1、Jag1)mRNA表达，采用Western blot法检测Notch信号通路相关蛋白的表达。 结果 与LPS组比较，DIDS高剂量组可以明显提高小鼠存活率，差异有高度统计学意义(P < 0.01);与空白对照组比较，LPS组TNF-α、IL-1β和IL-6的 mRNA均显著升高，差异均有统计学意义(均P < 0.05);与LPS组比较，DIDS高剂量组TNF-α、IL-1β和IL-6的mRNA显著降低，差异均有统计学意义(均P < 0.05)。与LPS组比较，DIDS高剂量组Notch信号相关基因Notch1、Notch3、Dll 1及Jag 1的mRNA表达水平升高，Dll 1和Jag 1相关分子蛋白水平升高，差异均有统计学意义(均P < 0.05)。 结论 DIDS能够明显减轻脓毒症小鼠的心肌炎性反应，其机制可能与调控Notch信号通路有关。
[关键词] 4，4′-二异硫氰基芪-2，2′-二磺酸;炎性反应;脓毒症;Notch信号通路
[中图分类号] R967 [文献标识码] A [文章编号] 1673-7210(2020)01(c)-0004-05
Effect and mechanism of 4，4′-diisothiocyanostilbene-2，2′-disulfonic acid on myocardial inflammatory response in sepsis mice
WANG Yan XIA Lin CAI Huazhong XIONG Yuyun
1.Department of Pharmacy， Zhenjiang First People′s Hospital， Jiangsu Province， Zhenjiang 212001， China; 2.Department of Clinical Laboratory， Affiliated Hospital of Jiangsu University， Jiangsu Province， Zhenjiang 212001， China; 3.Department of Emergency， Affiliated Hospital of Jiangsu University， Jiangsu Province， Zhenjiang 212001， China
[Abstract] Objective To investigate the effect and mechanism of 4，4′-diisothiocyanostilbene-2，2′-disulfonic acid (DIDS) on myocardial inflammatory response in sepsis mice. Methods A total of 52 male BALB/c mice were divided into 4 groups by random number table method， namely blank control group， lipopolysaccharide (LPS) group， low dose DIDS group and high dose DIDS group， with 13 mice in each group. For 3 days of pretreatment， the low dose DIDS group and the high dose DIDS group were intraperitoneally injected with 7 and 14 mg/kg DIDS every day， respectively， while the blank control group and the LPS group were given the same amount of phosphate buffer salt solution (PBS) intraperitoneally every day. After 2 h of pretreatment on the third day， PBS was injected intraperitoneally into the blank control group， and LPS 10 mg/kg was injected intraperitoneally into the other three groups to establish a mouse sepsis model. After the model was created successfully， the survival status of 10 mice in each group were observed for 48 h， and the survival rates were calculated. In addition， 3 mice in each group were sacrificed after 4 h， and their heart tissues were taken. The mRNA expression of inflammatory factors [including tumor necrosis factor-α (TNF-α)， interleukin-1β (IL-1β) and interleukin-6 (IL-6)] and genes related to Notch signaling pathway (including Notch1-4， Dll1 and Jag1) were detected by real-time PCR. Western blot was used to detect the expression of proteins related to the Notch signaling pathway. Results Compared with the LPS group， the survival rate of mice in the high dose DIDS group was significantly improved， with a highly statistically significant difference (P < 0.01). Compared with the blank control group， the mRNA levels of TNF-α，IL-1β and IL-6 in the LPS group were significantly increased， with statistically significant differences (all P < 0.05). Compared with the LPS group， the mRNA levels of TNF-α，IL-1β and IL-6 in the high dose DIDS group were significantly decreased， with statistically significant differences (all P < 0.05). Compared with the LPS group， mRNA expression levels of Notch1， Notch3， Dll 1 and Jag 1 of Notch signaling related genes in the high-dose DIDS group were increased， and protein levels of Dll 1 and Jag 1 were increased， with statistically significant differences (all P < 0.05). Conclusion DIDS can significantly reduce the myocardial inflammatory response in sepsis mice， and its mechanism may be related to the regulation of Notch signaling pathway., http://www.100md.com(王艳 夏琳 蔡华忠 熊御)
[关键词] 4，4′-二异硫氰基芪-2，2′-二磺酸;炎性反应;脓毒症;Notch信号通路
[中图分类号] R967 [文献标识码] A [文章编号] 1673-7210(2020)01(c)-0004-05
Effect and mechanism of 4，4′-diisothiocyanostilbene-2，2′-disulfonic acid on myocardial inflammatory response in sepsis mice
WANG Yan XIA Lin CAI Huazhong XIONG Yuyun
1.Department of Pharmacy， Zhenjiang First People′s Hospital， Jiangsu Province， Zhenjiang 212001， China; 2.Department of Clinical Laboratory， Affiliated Hospital of Jiangsu University， Jiangsu Province， Zhenjiang 212001， China; 3.Department of Emergency， Affiliated Hospital of Jiangsu University， Jiangsu Province， Zhenjiang 212001， China
[Abstract] Objective To investigate the effect and mechanism of 4，4′-diisothiocyanostilbene-2，2′-disulfonic acid (DIDS) on myocardial inflammatory response in sepsis mice. Methods A total of 52 male BALB/c mice were divided into 4 groups by random number table method， namely blank control group， lipopolysaccharide (LPS) group， low dose DIDS group and high dose DIDS group， with 13 mice in each group. For 3 days of pretreatment， the low dose DIDS group and the high dose DIDS group were intraperitoneally injected with 7 and 14 mg/kg DIDS every day， respectively， while the blank control group and the LPS group were given the same amount of phosphate buffer salt solution (PBS) intraperitoneally every day. After 2 h of pretreatment on the third day， PBS was injected intraperitoneally into the blank control group， and LPS 10 mg/kg was injected intraperitoneally into the other three groups to establish a mouse sepsis model. After the model was created successfully， the survival status of 10 mice in each group were observed for 48 h， and the survival rates were calculated. In addition， 3 mice in each group were sacrificed after 4 h， and their heart tissues were taken. The mRNA expression of inflammatory factors [including tumor necrosis factor-α (TNF-α)， interleukin-1β (IL-1β) and interleukin-6 (IL-6)] and genes related to Notch signaling pathway (including Notch1-4， Dll1 and Jag1) were detected by real-time PCR. Western blot was used to detect the expression of proteins related to the Notch signaling pathway. Results Compared with the LPS group， the survival rate of mice in the high dose DIDS group was significantly improved， with a highly statistically significant difference (P < 0.01). Compared with the blank control group， the mRNA levels of TNF-α，IL-1β and IL-6 in the LPS group were significantly increased， with statistically significant differences (all P < 0.05). Compared with the LPS group， the mRNA levels of TNF-α，IL-1β and IL-6 in the high dose DIDS group were significantly decreased， with statistically significant differences (all P < 0.05). Compared with the LPS group， mRNA expression levels of Notch1， Notch3， Dll 1 and Jag 1 of Notch signaling related genes in the high-dose DIDS group were increased， and protein levels of Dll 1 and Jag 1 were increased， with statistically significant differences (all P < 0.05). Conclusion DIDS can significantly reduce the myocardial inflammatory response in sepsis mice， and its mechanism may be related to the regulation of Notch signaling pathway., http://www.100md.com(王艳 夏琳 蔡华忠 熊御)