白藜芦醇通过Toll样受体4通路对高糖致H9C2心肌细胞损伤的作用研究
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摘要
目的研究白藜芦醇通过Toll样受体4(TLR4)通路对高糖致H9C2心肌细胞损伤的保护作用。方法 100 mmol/L高糖处理H9C2心肌细胞48 h以建立H9C2心肌细胞高糖损伤模型,实验分为对照组、高糖处理组、白藜芦醇+高糖处理组,酶联免疫吸附法(ELISA)检测各组乳酸脱氢酶(LDH)、丙二醛(MDA)和超氧化物歧化酶(SOD)含量,CCK8检测细胞增殖,AnnexinV/PI检测细胞凋亡,蛋白质印迹法检测TLR4蛋白的表达水平。结果高糖处理组较对照组LDH、MDA、TLR4表达明显升高,SOD明显降低,细胞增殖被抑制,细胞凋亡不明显;白藜芦醇预处理后LDH、MDA明显下降,TLR4表达有所降低,细胞增殖和细胞凋亡有所改善。结论高糖心肌损伤可引起TLR4表达增加,白藜芦醇可能在一定程度上通过抑制TLR4表达对糖尿病心肌细胞发挥保护作用。
Objective To study the protective effect of resveratrol against high glucose induced injury of H9 C2 cardiomyocytes via the Toll-like receptor 4(TLR4) pathway. Methods H9 C2 cardiomyocytes were treated with high levelglucose(100 mmol/L)for 48 h to establish the high glucose induced injury model of H9 C2 cardiomyocytes. This study included several groups: the control group, high glucose group, and Resveratrol + high glucose group. ELISA was used to detect the levels of lactate dehydrogenase(LDH), malondialdehyde(MDA) and superoxide dismutase(SOD),CCK8 assay was used to examinethe cell proliferation. Cell apoptosis was detected by Annexin V/PI assay. The protein expression level of TLR4 was measured with Western-blotting. Results Compared with the control group, the high glucose group showed significantly increased expression of LDH, MDA and TLR4, lowered level of SOD, sup-pression of cell proliferation, and no obvious signs of cell apoptosis. Pretreatment with resveratrol led to significantly decreasedlevel of LDH and MDA, lowered expression of TLR4, and improvements in cell proliferation and apoptosis.Conclusion High glucose induced myocardial injury result in increased TLR4 expression. Resveratrol may offer some protection for diabetic cardiomyocytes by inhibiting TLR4 expression.
Objective To study the protective effect of resveratrol against high glucose induced injury of H9 C2 cardiomyocytes via the Toll-like receptor 4(TLR4) pathway. Methods H9 C2 cardiomyocytes were treated with high levelglucose(100 mmol/L)for 48 h to establish the high glucose induced injury model of H9 C2 cardiomyocytes. This study included several groups: the control group, high glucose group, and Resveratrol + high glucose group. ELISA was used to detect the levels of lactate dehydrogenase(LDH), malondialdehyde(MDA) and superoxide dismutase(SOD),CCK8 assay was used to examinethe cell proliferation. Cell apoptosis was detected by Annexin V/PI assay. The protein expression level of TLR4 was measured with Western-blotting. Results Compared with the control group, the high glucose group showed significantly increased expression of LDH, MDA and TLR4, lowered level of SOD, sup-pression of cell proliferation, and no obvious signs of cell apoptosis. Pretreatment with resveratrol led to significantly decreasedlevel of LDH and MDA, lowered expression of TLR4, and improvements in cell proliferation and apoptosis.Conclusion High glucose induced myocardial injury result in increased TLR4 expression. Resveratrol may offer some protection for diabetic cardiomyocytes by inhibiting TLR4 expression.
引文
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[4] Echem C,Bomfim GF,Ceravolo GS,et al. Anti-toll like receptor4(TLR4)therapy diminishes cardiac remodeling regardless of changes in blood pressure in spontaneously hypertensive rats(SHR)[J]. Int J Cardiol,2015,187:243-245.
[5] Topkara VK,Evans S,Zhang W,et al. Therapeutic targeting of innate immunity in the failing heart[J]. J Mol Cell Cardiol,2011,51(4):594-599.
[6] Liu ZW,Wang JK,Qiu C,et al. Matrine pretreatment improves cardiac function in rats with diabetic cardiomyopathy via suppressing ROS/TLR4 signaling pathway[J]. Acta Pharmacol Sin,2015,36(3):323-333.
[7] Al-Hussaini H,Kilarkaje N. Trans-resveratrol mitigates type 1 diabetes-induced oxidative DNA damage and accumulation of advanced glycation end products in glomeruli and tubules of rat kidneys[J]. Toxicol Appl Pharmacol,2018,339:97-109.
[8] Stratmann B,Stork I,Tschoepe D. Diabetic cardiomyopathy a type of coronary artery disease?[J]. Internist(Berl),2008,49(4):436-440.
[9] Wan A,Rodrigues B. Endothelial cell-cardiomyocyte crosstalk in diabetic cardiomyopathy[J]. Cardiovasc Res,2016,111(3):172-183.
[10] Frantz S,Ertl G,Bauersachs J. Mechanisms of disease:Toll like receptors in cardiovascular disease[J]. Nat Clin Pract Cardiovasc Med,2007,4(8):444-454.
[11] Heineke J,Molkentin JD. Regulation of cardiac hypertrophy by intracellular signalling pathways[J]. Nat Rev Mol Cell Biol,2006,7(8):589-600.
[12] Bernardo BC,Weeks KL,Pretorius L,et al. Molecular distinction between physiological and pathological cardiac hypertrophy:3xperimental findings and therapeutic strategies[J]. Pharmacol Ther,2010,128(1):191-227.
[13] Tao A,Song J,Lan T,et al. Cardiomyocyte-fibroblast interaction contributes to diabetic cardiomyopathy in mice:role of HMGB1/TLR4/IL-33 axis[J]. Biochim Biophy Acta,2015,1852(10PtA):2075-2085.
[2] Bugger H,Abel ED. Molecular mechanisms of diabetic cardiomyopathy[J]. Diabetologia,2014,57(4):660-671.
[3] Jia G,deMarco VG,Sowers JR. Insulin resistance and hyperinsulinaemia in diabetic cardiomyopathy[J]. Nat Rev Endocrinol,2016,12(3):144-153.
[4] Echem C,Bomfim GF,Ceravolo GS,et al. Anti-toll like receptor4(TLR4)therapy diminishes cardiac remodeling regardless of changes in blood pressure in spontaneously hypertensive rats(SHR)[J]. Int J Cardiol,2015,187:243-245.
[5] Topkara VK,Evans S,Zhang W,et al. Therapeutic targeting of innate immunity in the failing heart[J]. J Mol Cell Cardiol,2011,51(4):594-599.
[6] Liu ZW,Wang JK,Qiu C,et al. Matrine pretreatment improves cardiac function in rats with diabetic cardiomyopathy via suppressing ROS/TLR4 signaling pathway[J]. Acta Pharmacol Sin,2015,36(3):323-333.
[7] Al-Hussaini H,Kilarkaje N. Trans-resveratrol mitigates type 1 diabetes-induced oxidative DNA damage and accumulation of advanced glycation end products in glomeruli and tubules of rat kidneys[J]. Toxicol Appl Pharmacol,2018,339:97-109.
[8] Stratmann B,Stork I,Tschoepe D. Diabetic cardiomyopathy a type of coronary artery disease?[J]. Internist(Berl),2008,49(4):436-440.
[9] Wan A,Rodrigues B. Endothelial cell-cardiomyocyte crosstalk in diabetic cardiomyopathy[J]. Cardiovasc Res,2016,111(3):172-183.
[10] Frantz S,Ertl G,Bauersachs J. Mechanisms of disease:Toll like receptors in cardiovascular disease[J]. Nat Clin Pract Cardiovasc Med,2007,4(8):444-454.
[11] Heineke J,Molkentin JD. Regulation of cardiac hypertrophy by intracellular signalling pathways[J]. Nat Rev Mol Cell Biol,2006,7(8):589-600.
[12] Bernardo BC,Weeks KL,Pretorius L,et al. Molecular distinction between physiological and pathological cardiac hypertrophy:3xperimental findings and therapeutic strategies[J]. Pharmacol Ther,2010,128(1):191-227.
[13] Tao A,Song J,Lan T,et al. Cardiomyocyte-fibroblast interaction contributes to diabetic cardiomyopathy in mice:role of HMGB1/TLR4/IL-33 axis[J]. Biochim Biophy Acta,2015,1852(10PtA):2075-2085.