新型芹菜素纳米脂质体对糖尿病心肌病大鼠心肌细胞凋亡的影响
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摘要
目的观察新型芹菜素纳米脂质体对糖尿病心肌病大鼠心肌细胞凋亡水平的影响。方法采用薄膜水合结合超声分散技术制备新型芹菜素纳米脂质体并进行质量考察。将健康雄性SD大鼠60只,随机分成正常对照组、模型对照组、空白脂质体组及芹菜素脂质体组,每组15只。采用大鼠腹腔注射链脲佐菌素(70 mg·kg-1)建立糖尿病模型,2周后,芹菜素脂质体组尾静脉给予芹菜素纳米脂质体30μg·kg-1,空白脂质体组尾静脉给予等剂量空白纳米脂质体,模型对照组及正常对照组给予同等剂量0.9%氯化钠溶液。12周之后处死大鼠,取出心肌组织,分别采用TUNEL凋亡染色测定大鼠心肌细胞凋亡指数,Western blotting分析各组大鼠心肌Bax及Bcl-2蛋白表达。结果 TUNEL凋亡染色显示,与正常对照组比较,模型对照组及空白脂质体组大鼠心肌细胞凋亡指数显著升高(P<0.05);芹菜素脂质体组凋亡指数显著下降(P<0.05)。Western blotting分析显示,与正常对照组比较,模型对照组及空白脂质体组大鼠心肌细胞Bax含量显著升高(P<0.01),Bcl-2含量显著下降(P<0.01);与模型对照组及空白纳米脂质体组比较,芹菜素脂质体组心肌细胞Bax含量显著下降(P<0.01),Bcl-2含量显著升高(P<0.01)。结论芹菜素纳米脂质体能够通过调节细胞凋亡相关蛋白Bcl-2/Bax途径,降低糖尿病心肌病大鼠心肌细胞凋亡水平。
Objective To investigate the effect of apigenin-loaded nanoliposomes on myocardial cells apoptosis induced by diabetic cardiomyopathy. Methods The apigenin-loaded nanoliposomes were prepared by film hydration and ultrasonic dispersion technology and their quality inspections were also investigated. Sixty SD rats were randomly divided into normal control group,model control group,blank and apigenin-loaded nano-liposomes group. Rat model of typeⅠdiabetes was induced by single intraperitoneal injection of STZ. After two weeks of STZ injection,the model rats were used for this study. The rats of apigenin loaded nano-liposomes treatment group were treated with apigenin loaded nano-liposomes via caudal vein administration for 12 weeks( three times a week). And the rats of normal control group and model control group were administrated equivalent volume of 0.9% sodium chloride solution. After treatment for 12 weeks,the experimental animals were sacrificed and their hearts were harvested after 0.9% sodium chloride solution perfusion. The myocardial cell apoptosis index was detected by TUNEL staining and the expressions of Bcl-2 and Bax,which were related to the cell apoptosis,were examined by Western blotting analysis.Results TUNEL staining showed the apoptosis index in the rats of diabetes and blank nano-liposomes groups were significant higher than the normal control group( P<0.05). However,there was significant decrease of apoptosis index in the diabetes rats after the treatment of apigenin loaded nano-lipsomes( P < 0.05). Moreover,Western blotting analysis confirmed that there was significant lower expression of Bcl-2 and higher expression of Bax in the hearts of model control group and blank nano-liposomes treated rats compared with normal control rats( P < 0. 01). Compared with the diabetes and blank nano-liposomes group,the apigenin loaded nano-liposomes treated groups showed higher expression of anti-apoptosis protein Bcl-2( P < 0. 01) and lower expression of pro-apoptosis protein Bax( P<0.01). Conclusion Apigenin loaded nano-liposomes have reduced the apoptosis of myocardial cells in diabetic cardiomyopathy rats via the Bcl-2/Bax pathway.
Objective To investigate the effect of apigenin-loaded nanoliposomes on myocardial cells apoptosis induced by diabetic cardiomyopathy. Methods The apigenin-loaded nanoliposomes were prepared by film hydration and ultrasonic dispersion technology and their quality inspections were also investigated. Sixty SD rats were randomly divided into normal control group,model control group,blank and apigenin-loaded nano-liposomes group. Rat model of typeⅠdiabetes was induced by single intraperitoneal injection of STZ. After two weeks of STZ injection,the model rats were used for this study. The rats of apigenin loaded nano-liposomes treatment group were treated with apigenin loaded nano-liposomes via caudal vein administration for 12 weeks( three times a week). And the rats of normal control group and model control group were administrated equivalent volume of 0.9% sodium chloride solution. After treatment for 12 weeks,the experimental animals were sacrificed and their hearts were harvested after 0.9% sodium chloride solution perfusion. The myocardial cell apoptosis index was detected by TUNEL staining and the expressions of Bcl-2 and Bax,which were related to the cell apoptosis,were examined by Western blotting analysis.Results TUNEL staining showed the apoptosis index in the rats of diabetes and blank nano-liposomes groups were significant higher than the normal control group( P<0.05). However,there was significant decrease of apoptosis index in the diabetes rats after the treatment of apigenin loaded nano-lipsomes( P < 0.05). Moreover,Western blotting analysis confirmed that there was significant lower expression of Bcl-2 and higher expression of Bax in the hearts of model control group and blank nano-liposomes treated rats compared with normal control rats( P < 0. 01). Compared with the diabetes and blank nano-liposomes group,the apigenin loaded nano-liposomes treated groups showed higher expression of anti-apoptosis protein Bcl-2( P < 0. 01) and lower expression of pro-apoptosis protein Bax( P<0.01). Conclusion Apigenin loaded nano-liposomes have reduced the apoptosis of myocardial cells in diabetic cardiomyopathy rats via the Bcl-2/Bax pathway.
引文
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[15]HU J,LI Z,XU L T,et al.Protective effect of apigenin on ischemia/reperfusion injury of the isolated rat heart[J].Cardiovas Toxicol,2015,15(3):241-249.
[16]ZHAO Y Z,ZHANG M,TIAN X Q,et al.Using basic fibroblast growth factor nanoliposome combined with ultrasound-introduced technology to early intervene the diabetic cardiomyopathy[J].Int J Nanomedic,2016,11(4):675-686.
[17]XIE Z,KOYAMA T,SUZUKI J,et al.Coronary reperfusion following ischemia:different expression of bcl-2 and bax proteins,and cardiomyocyte apoptosis[J].Japan Heart J,2001,42(6):759-770.
[2]ZIMMET P,ALBERTI K G,SHAW J.Global and societal implications of the diabetes epidemic[J].Nature,2001,414(6865):782-787.
[3]KANNEL W B,MCGEE D L.Diabetes and cardiovascular risk factors:the Framingham study[J].Circulation,1979,59(1):8-13.
[4]CAI L,KANG Y J.Cell death and diabetic cardiomyopathy[J].Cardiovasc Toxicol,2003,3(3):219-228.
[5]刘俊法.芹菜素对糖尿病大鼠降血糖、调节血脂和抗氧化能力的影响[J].中药药理与临床,2014,5(1):44-47.
[6]ZHANG J,LIU D,HUANG Y,et al.Biopharmaceutics classification and intestinal absorption study of apigenin[J].Int J Pharmac,2012,436(1/2):311-317.
[7]CASTOLDI A,HERR C,NIEDERSTRASSER J,et al.Calcifediol-loaded liposomes for local treatment of pulmonary bacterial infections[J].Eur J Pharmac Biopharmaceutics,2017,118(1):62-67.
[8]JORAHOLMEN M W,BASNET P,ACHARYA G,et al.PEGylated liposomes for topical vaginal therapy improve delivery of interferon alpha[J].Eur J Pharmac Biopharmaceutics,2017,113(1):132-139.
[9]JIN X,YANG Q,ZHANG Y.Synergistic apoptotic effects of apigenin TPGS liposomes and tyroservatide:implications for effective treatment of lung cancer[J].Int J Nanomedicine,2017,12(5):109-118.
[10]ZHAO Y Z,ZHANG M,WONG H L,et al.Prevent diabetic cardiomyopathy in diabetic rats by combined therapy of a FGF-loaded nanoparticles and ultrasound-targeted microbubble destruction technique[J].J Control Rel Soc,2016,223(1):11-21.
[11]CAI L,LI W,WANG G,et al.Hyperglycemia-induced apoptosis in mouse myocardium:mitochondrial cytochrome C-mediated caspase-3 activation pathway[J].Diabetes,2002,51(6):1938-1948.
[12]ZOU M H,XIE Z.Regulation of interplay between autophagy and apoptosis in the diabetic heart:new role of AMPK[J].Autophagy,2013,9(4):624-625.
[13]LEFORT E C,BLAY J.Apigenin and its impact on gastrointestinal cancers[J].Molecular Nutrit Food Res,2013,57(1):126-144.
[14]JUNG U J,CHO Y Y,CHOI M S.Apigenin ameliorates dyslipidemia,hepatic steatosis and insulin resistance by modulating metabolic and transcriptional profiles in the liver of high-fat diet-induced obese mice[J].Nutrients,2016,8(5):E305.
[15]HU J,LI Z,XU L T,et al.Protective effect of apigenin on ischemia/reperfusion injury of the isolated rat heart[J].Cardiovas Toxicol,2015,15(3):241-249.
[16]ZHAO Y Z,ZHANG M,TIAN X Q,et al.Using basic fibroblast growth factor nanoliposome combined with ultrasound-introduced technology to early intervene the diabetic cardiomyopathy[J].Int J Nanomedic,2016,11(4):675-686.
[17]XIE Z,KOYAMA T,SUZUKI J,et al.Coronary reperfusion following ischemia:different expression of bcl-2 and bax proteins,and cardiomyocyte apoptosis[J].Japan Heart J,2001,42(6):759-770.