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目的分析糖尿病心肌病SD大鼠心肌组织中脾酪氨酸激酶(Syk)、c-Jun氨基末端激酶(JNK)和核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体的表达水平,并探讨三者的作用关系。方法清洁级雄性SD大鼠随机分为对照(Ctrl)组和模型(DCM)组,DCM组采用链脲佐菌素(STZ)一次性腹腔注射方法造模,Ctrl组注射同等剂量的柠檬酸缓冲液。每周监测2组随机血糖和体质量,直至STZ注射后20周,处死取其心脏。大鼠H9c2心肌细胞随机分为正常糖处理(NG)组、高糖处理(HG)组、Syk抑制剂对照(BAY)组,Syk抑制剂高糖(HG+BAY)组。Western blot方法在蛋白水平检测大鼠心肌组织和H9c2心肌细胞中Syk、JNK的磷酸化及NLRP3的表达;逆转录聚合酶链反应(RT-PCR)方法在mRNA水平检测NLRP3、半胱氨酸天门冬氨酸特异性蛋白酶1(caspase-1)及白细胞介素(IL)-1β的表达。结果与Ctrl组相比,DCM组大鼠的随机血糖水平显著升高,体质量显著降低(P<0.05),DCM组大鼠心肌组织中p-Syk、p-JNK及NLRP3的蛋白表达明显增加(P<0.05),且NLRP3、caspase-1和IL-1βmRNA的表达也显著增加(P<0.05)。BAY处理后显著抑制了高糖诱导的H9c2心肌细胞中NLRP3、caspase-1、IL-1βmRNA的表达和p-JNK、NLRP3蛋白的表达(P<0.05)。结论 Syk诱导的JNK磷酸化及NLRP3炎症小体的活化在糖尿病心肌病的发病机制中发挥关键作用。
Objective To analyze the expressions of Spleen tyrosine kinase (Syk), c-Jun N-terminal kinase (JNK) and Nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome in SD rats with diabetic cardiomyopathy The level of expression, and explore the relationship between the three. Methods Clean male Sprague-Dawley rats were randomly divided into control group (Ctrl group) and model group (DCM group). DCM group was given intraperitoneal injection of streptozotocin (STZ). The rats in Ctrl group were injected with the same dose of citrate buffer liquid. Two groups of randomized blood glucose and body weight were monitored weekly until 20 weeks after STZ injection, sacrificed for taking their heart. Rat H9c2 cardiomyocytes were randomly divided into normal glucose treatment (NG) group, high glucose treatment (HG) group, Syk inhibitor control (BAY) group and Syk inhibitor high glucose (HG + BAY) group. Western blot was used to detect the phosphorylation of Syk and JNK and the expression of NLRP3 in rat cardiac myocytes and H9c2 cardiomyocytes. RT-PCR was used to detect the expression of NLRP3, cysteine Tianmen The expression of caspase-1 and IL-1β. Results Compared with the Ctrl group, the DCM group had a significantly increased random blood glucose level and a significant decrease in body weight (P <0.05). The protein expression of p-Syk, p-JNK and NLRP3 in the DCM group was significantly increased (P <0.05), and the expression of NLRP3, caspase-1 and IL-1βmRNA also increased significantly (P <0.05). BAY significantly inhibited the expression of NLRP3, caspase-1 and IL-1βmRNA and the expressions of p-JNK and NLRP3 in H9c2cells induced by high glucose (P <0.05). Conclusions Syk-induced JNK phosphorylation and NLRP3 inflammasome activation play a key role in the pathogenesis of diabetic cardiomyopathy.