KLF4在糖尿病小鼠心肌组织的表达及通心络胶囊干预作用研究
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摘要
目的探讨糖尿病小鼠心肌组织Krüppel样因子4(KLF4)表达及通心络(TXL)对其干预作用。方法 KK/Upj-Ay小鼠40只,随机分为糖尿病模型组、TXL低、中、高剂组,每组10只,另设C57BL/6(C57)小鼠10只作为对照组。按照分组给予不同剂量处理因素,疗程3个月,测定各组空腹血糖(FBG)、糖化血红蛋白(Hb A1c)、甘油三酯(TG)、总胆固醇(TC),空腹胰岛素(FINS)、计算胰岛素抵抗指数(HOMA-IR);放射免疫法测定各组血清TNF-α、IL-6水平;HE染色观察心肌组织病理变化;Real time-PCR和Western blot测定心肌组织KLF4 m RNA和蛋白表达;Western blot测定心肌组织核因子-κB(nuclear factor kappa B,NF-κB)核蛋白表达。结果与对照组比较,模型组FBG、Hb A1c、TG、TC、FINS、HOMA-IR及TNF-α、IL-6水平明显增高,KLF4 m RNA和蛋白表达明显降低,NF-κB核蛋白表达明显升高(P<0.01),通心络胶囊能够明显减轻心肌组织病理损伤,降低FINS、TG、TC、HOMA-IR、TNF-α、IL-6水平及NF-κB核蛋白表达,同时上调KLF4表达(P<0.05),但不影响FBG、Hb A1c(P>0.05)。结论 KLF4可能参与了糖尿病心肌病的发生发展。通心络胶囊可减轻糖尿病心肌病理损伤及改善心功能,其机制可能与上调KLF4表达及抑制NF-κB炎症信号通路的激活有关。
Aim To observe the expressions of KLF4 in the myocardium of diabetic mice and their changes under Tongxinluo capsule intervention. Methods Forty KK / Upj-Ay mice were randomly divided into diabetic model group( n = 10) and diabetic model with Tongxinluo( TXL low,middle,high) groups( n = 10,respectively). C57 BL /6 mice were selected as control group(n = 10). At the end of the 3th month the mice were sacrificed and were weighed. The fasting blood-glucose(FBG),glycosylated hemoglobin(Hb A1c),triglyceride(TG),total cholesterol( TC) and insulin( FINS) were measured to calculate HOMA-IR. Radioimmunoassay was used to measure serum TNF-α and IL-6. The pathological changes in the myocardium of mice were observed by HE staining. KLF4 m RNA was examined by Real-time PCR,while KLF4 and NF-κB protein were measured by Western blot. Results Compared to the control group,FBG,Hb A1 c,TG,TC,FINS,HOMA-IR,TNF-α and IL-6 in model group were markedly increased; the expressions of myocardial KLF4 were markedly decreased and the expression of nuclear NF-κB protein were markedly increased( P < 0. 01).Tongxinluo capsule could significantly reduce myocardial pathological damage,FINS,TG,TC,TNF-α,IL-6level and the expression of nuclear NF-κB protein and up-regulate the expression of KLF4( P < 0. 05),but had no effect on FBG and Hb A1c( P > 0. 05). Conclusions KLF4 may be involved in the development of myocardial injury during diabetes. Tongxinluo capsule can ameliorate the myocardial damage and improve the function of diabetic myocardium by up-regulating the expression of KLF4 and inhibiting the activation of NF-κB inflammatory signal pathway.
Aim To observe the expressions of KLF4 in the myocardium of diabetic mice and their changes under Tongxinluo capsule intervention. Methods Forty KK / Upj-Ay mice were randomly divided into diabetic model group( n = 10) and diabetic model with Tongxinluo( TXL low,middle,high) groups( n = 10,respectively). C57 BL /6 mice were selected as control group(n = 10). At the end of the 3th month the mice were sacrificed and were weighed. The fasting blood-glucose(FBG),glycosylated hemoglobin(Hb A1c),triglyceride(TG),total cholesterol( TC) and insulin( FINS) were measured to calculate HOMA-IR. Radioimmunoassay was used to measure serum TNF-α and IL-6. The pathological changes in the myocardium of mice were observed by HE staining. KLF4 m RNA was examined by Real-time PCR,while KLF4 and NF-κB protein were measured by Western blot. Results Compared to the control group,FBG,Hb A1 c,TG,TC,FINS,HOMA-IR,TNF-α and IL-6 in model group were markedly increased; the expressions of myocardial KLF4 were markedly decreased and the expression of nuclear NF-κB protein were markedly increased( P < 0. 01).Tongxinluo capsule could significantly reduce myocardial pathological damage,FINS,TG,TC,TNF-α,IL-6level and the expression of nuclear NF-κB protein and up-regulate the expression of KLF4( P < 0. 05),but had no effect on FBG and Hb A1c( P > 0. 05). Conclusions KLF4 may be involved in the development of myocardial injury during diabetes. Tongxinluo capsule can ameliorate the myocardial damage and improve the function of diabetic myocardium by up-regulating the expression of KLF4 and inhibiting the activation of NF-κB inflammatory signal pathway.
引文
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[12]游琼,吴铿,涂焰明,等.柚皮苷调控心肌核因子NF-κB炎症信号通路对糖尿病心肌病大鼠防治作用[J].中国免疫学杂志,2012,29(2):121-4.You Q,Wu K,Tu Y M,et al.Preventive effect of naringin on cardiac NF-κB inflammatory signal pathway in diabetic cardiomyopathy rats[J].Chin J Immu,2012,29(2):121-4.
[13]Wang B,Yang Q,Bai W W,et al.Tongxinluo protects against pressure overload-induced heart failure in mice involving VEGF/Akt/e NOS pathway activation[J].PLo S One,2014,9(6):e98047.
[14]Li X D,Yang Y J,Cheng Y T,et al.Protein kinase A-mediated cardioprotection of Tongxinluo relates to the inhibition of myocardial inflammation,apoptosis,and edema in reperfused swine hearts[J].Chin Med J(Engl),2013,126(8):1469-79.
[2]董世芬,洪缨,汪瑞祺,等.小檗碱对实验性2型糖尿病心肌病大鼠模型心脏保护作用研究[J].中国药理学通报,2013,29(9):1216-21.Dong S F,Hong Y,Wang R Q,et al.Berberine attenuates cardiac dysfunction in experimental type 2 diabetic cardiomyopathy rat model[J].Chin Pharmacol Bull,2013,29(9):1216-21.
[3]张哲,赵志英,王超.KLF4通过上调Mfn2表达减轻棕榈酸所致L6骨骼肌细胞的胰岛素抵抗[J].中国老年学杂志,2014,34(21):6076-9.Zhang Z,Zhao Z Y,Wang C.KLF4 ameliorates palmitate-induced insulin resistance partially by up-regulating Mfn2 expression in L6skeleton muscle cells[J].Chin J Gerontol,2014,34(21):6076-9.
[4]Yoshida T,Yamashita M,Horimai C,et al.Kruppel-like factor 4protein regulates isoproterenol-induced cardiac hypertrophy by modulating myocardin expression and activity[J].J Biol Chem,2014,289(38):26107-18.
[5]Gao D,Madi M,Ding C,et al.Interleukin-1βmediates macrophage-induced impairment of insulin signaling in human primary adipocytes[J].Am J Physiol Endocrinol Met,2014,307(3):E289-304.
[6]Lorenzo O,Picatoste B,Ares-Carrasco S,et al.Potential role of nuclear factorκB in diatetic cardiomyopathy[J].Mediators Inflamm,2011,2011:652097.
[7]Min W,Bin Z W,Quan Z B,et al.The signal transduction pathway of PKC/NF-κB/c-fos may be involved in the influence of high glucose on the cardiomyocytes of neonatal rats[J].Cardiovasc Diabetol,2009,8:8.
[8]高惠宽,吴永全.核因子κB与糖尿病心肌病[J].中华临床医师杂志(电子版),2012,6(21):6860-1.Gao H K,Wu Y Q.Nuclear factorκB and diabetic cardiomyopathy[J].Chin J Clinicians(Electronic Edition),2012,6(21):6860-1.
[9]Lin Z S,Chu H C,Yen Y C,et al.Krüppel-like factor 4,a tumor suppressor in hepatocellular carcinoma cells reverts epithelial mesenchymal transition by suppressing slug expression[J].PLo S One,2012,7(8):e43593.
[10]Shen B,Smith R S Jr,Hsu Y T,et al.Kruppel-like factor 4 is a novel mediator of Kallistatin in inhibiting endothelial inflammation via increased endothelial nitric-oxide synthase expression[J].J Biol Chem,2009,284(51):35471-8.
[11]Yoshida T,Yamashita M,Horimai C,et al.Deletion of Krüppellike factor 4 in endothelial and hematopoietic cells enhances neointimal formation following vascular injury[J].J Am Heart Assoc,2014,3(1):e000622.
[12]游琼,吴铿,涂焰明,等.柚皮苷调控心肌核因子NF-κB炎症信号通路对糖尿病心肌病大鼠防治作用[J].中国免疫学杂志,2012,29(2):121-4.You Q,Wu K,Tu Y M,et al.Preventive effect of naringin on cardiac NF-κB inflammatory signal pathway in diabetic cardiomyopathy rats[J].Chin J Immu,2012,29(2):121-4.
[13]Wang B,Yang Q,Bai W W,et al.Tongxinluo protects against pressure overload-induced heart failure in mice involving VEGF/Akt/e NOS pathway activation[J].PLo S One,2014,9(6):e98047.
[14]Li X D,Yang Y J,Cheng Y T,et al.Protein kinase A-mediated cardioprotection of Tongxinluo relates to the inhibition of myocardial inflammation,apoptosis,and edema in reperfused swine hearts[J].Chin Med J(Engl),2013,126(8):1469-79.