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[ω-3多不饱和脂肪酸对糖尿病大鼠冠状动脉平滑肌细胞中钙库操纵性钙通道的影响]

[Effects of n-3 polyunsaturated fatty acid on store-operated calcium channels in coronary artery smooth muscle cells derived from diabetic rat].

作者信息

Tang X, Qian L L, Dang S P, Wu Y, Liu X Y, Zhang Z Y, Miu L F, Wang R X

机构信息

Department of Cardiology, Wuxi People's Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China (Tang Xu is working on the Department of Infectious Diseases, Wuxi People's Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China).

出版信息

Zhonghua Xin Xue Guan Bing Za Zhi. 2019 Aug 24;47(8):640-646. doi: 10.3760/cma.j.issn.0253-3758.2019.08.009.

Abstract

To investigate the impact of n-3 polyunsaturated fatty acid (n-3 PUFA) on function and expression of store-operated calcium channels (SOCC) in coronary artery smooth muscle cells (SMC) derived from diabetic rat. A total of 180 healthy male Sprague-Dawley (SD) rats were randomly divided into normal group (N, 45), placebo-treated diabetic group (D, 45), lose dose n-3 PUFA treated diabetic group (DL, 45) and high dose n-3 PUFAs treated diabetic group (DH, 45). Streptozotocin-induced diabetic rat animal model was established by two consecutive intraperitoneal injections. After modeling, rats in group DL and DH were treated with 10 mg·kg(-1)·d(-1) and 50 mg·kg(-1)·d(-1) n-3 PUFAs respectively per gavage for eight weeks. After eight weeks, rat coronary artery SMC was isolated by enzyme digestion. Changes of cytosolic calcium concentration in coronary artery SMC were examined by calcium fluorescence imaging technique, coronary artery tension was detected by myograph system, and protein expressions of SOCC on coronary artery SMC were measured by Western blot. SOCC induced ΔF340/F380 of group N, D, DL and DH were 0.425±0.023, 0.838±0.037, 0.342±0.052 and 0.364±0.045 respectively, which was significantly lower in group N, DL, DH than in group D (0.05). SOCC induced changes of tensions were 0.94±0.09, 1.95±0.18, 1.35±0.24 and 1.01±0.18 in the group N, D, DL and DH, respectively, which was significantly lower in group N and DH than in group D (0.05). Protein expressions of STIM1, Orai1 and TRPC1 were significantly higher in diabetic rat coronary SMC than in group N (0.05). STIM1 protein expressions were significantly lower in group DL and DH than in group D, and Orai1 and TRPC1 protein expressions were similar among group. Coronary artery tension, cytosolic calcium concentration and protein expressions of SOCC are higher in diabetic rat coronary artery SMC when compared with normal rats. n-3 PUFA intervention could downregulate the protein expression of SOCC, reduce cytosolic calcium concentration and coronary artery tension, and is protective to the diabetic injury in coronary artery.

摘要

研究n-3多不饱和脂肪酸(n-3 PUFA)对糖尿病大鼠冠状动脉平滑肌细胞(SMC)中储存-操作性钙通道(SOCC)功能及表达的影响。将180只健康雄性Sprague-Dawley(SD)大鼠随机分为正常组(N,45只)、安慰剂处理糖尿病组(D,45只)、低剂量n-3 PUFA处理糖尿病组(DL,45只)和高剂量n-3 PUFAs处理糖尿病组(DH,45只)。通过连续两次腹腔注射建立链脲佐菌素诱导的糖尿病大鼠动物模型。造模后,DL组和DH组大鼠分别按10 mg·kg⁻¹·d⁻¹和50 mg·kg⁻¹·d⁻¹的剂量经口灌胃给予n-3 PUFAs,持续8周。8周后,采用酶消化法分离大鼠冠状动脉SMC。采用钙荧光成像技术检测冠状动脉SMC胞质钙浓度变化,用肌张力描记系统检测冠状动脉张力,并用蛋白质印迹法检测冠状动脉SMC上SOCC的蛋白表达。N组、D组、DL组和DH组SOCC诱导的ΔF340/F380分别为0.425±0.023、0.838±0.037、0.342±0.052和0.364±0.045,N组、DL组、DH组均显著低于D组(P<0.05)。N组、D组、DL组和DH组SOCC诱导的张力变化分别为0.94±0.09、1.95±0.18、1.35±0.24和1.01±0.18,N组和DH组显著低于D组(P<0.05)。糖尿病大鼠冠状动脉SMC中STIM1、Orai1和TRPC1的蛋白表达显著高于N组(P<0.05)。DL组和DH组STIM1蛋白表达显著低于D组,Orai1和TRPC1蛋白表达在各组间相似。与正常大鼠相比,糖尿病大鼠冠状动脉SMC的冠状动脉张力、胞质钙浓度及SOCC蛋白表达更高。n-3 PUFA干预可下调SOCC蛋白表达,降低胞质钙浓度和冠状动脉张力,对冠状动脉糖尿病损伤具有保护作用。

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