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链脲佐菌素诱导的1型糖尿病小鼠肾脏COX-2/PGE2/EPs的表达[J]. 中山大学学报(医学科学版), 2015,36(1).
Expression of COX-2/PGE2/EPs in Kidney Tissues of STZ-induced Type 1 Diabetes[J]. Journal of Sun Yat-sen University (Medical Sciences), 2015, 36(1).
【目的】 探讨链脲佐菌素(STZ)诱导的1型糖尿病小鼠肾脏COX-2/PGE2/EPs的表达情况。【方法】 多次小剂量腹腔注射STZ诱导1型糖尿病小鼠模型,实验分为两组:正常对照组(NC)和糖尿病组(DM)。ELISA法检测血清和尿液中前列腺素E2(PGE2)的表达水平;RT-qPCR法检测肾组织环氧合酶-2(COX-2)、膜结合型前列腺素E2合成酶-1(mPGES-1)、各型EP受体(EP1、EP2、EP3、EP4)的mRNA表达水平;Westernblot 法检测相应蛋白表达水平。【结果】 随着周龄增加,DM组小鼠逐渐出现明显多饮、多尿、体质量下降等糖尿病消耗症状;与NC组小鼠相比,DM组小鼠空腹血糖(FBG)和24 h尿蛋白排泄量均显著增加(P<0.05);肾脏COX-2和EP4表达水平明显上调(P < 0.05)、而mPGES-1、EP1、EP2、EP3 mRNA表达无显著变化(P > 0.05);同时,DM组小鼠24 h尿PGE2定量显著高于NC组(P < 0.05)。【结论】1型糖尿病小鼠肾脏组织中,早期即可检测到COX-2和EP4表达上调,伴24 h尿PGE2定量增加,提示COX-2/PGE2/EP4信号轴激活可能参与了糖尿病肾病的发生发展。
【Objective】 To explore the expression of COX-2/PGE2/EPs in the kidney tissues of STZ-induced type 1 diabetes.【Methods】 Multiple low-dose STZ intraperitoneal injections were used to induce type 1 diabetes. Mice were divided into two groups: normal control (NC) and diabetes mellitus (DM). Urine and serum PGE2 concentrations were determined by the enzyme-linked immunoassay (ELISA). Real-time PCR was performed to assess the mRNA expression levels of cyclooxygenase-2 (COX-2), microsomal PGE synthase-1 (mPGES-1) and EP receptors (EP1, EP2, EP3, and EP4).Western blot was used to measure the protein expression levels. 【Results】 Associated with the increase of weeks, DM mice appeared polydipsia, polyuria, weight loss and the other diabetic symptoms. Fasting blood glucose level and 24h-urinary protein output were significantly elevated in DM mice comparing with the control groups (P < 0.05). Comparing to the NC group, the mRNA and protein expression levels of COX-2 and EP4 in the kidney tissues of DM mice were significantly up regulated, while the mRNA expression of mPGES–1, EP1, EP2, EP3 were not significantly altered between the two groups. Diabetes induced a significant rise in the excretion of PGE2 in the urine comparing with the control mice.【Conclusion】 In the early diabetes, renal COX-2 and EP4 expressions were up regulated, and the 24 h urine excretion rate of PGE2 was obviously increased, suggesting that the activation of COX-2/PGE2/EP4 pathway may play an important role in the occurrence and development of diabetic nephropathy.
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