针刺通过抑制肝脏氧化应激改善腹型肥胖型非酒精性脂肪肝病大鼠的肝脏脂质代谢
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摘要
目的:探讨针刺对腹型肥胖型非酒精性脂肪肝病(NAFLD)大鼠脂质代谢的影响及其机制。方法:雄性SD大鼠随机分为空白组11只和造模组30只,高脂饮食喂养12周,建立腹型肥胖合并NAFLD模型。空白组和造模组分别随机选取3只验证模型。最终造模成功的16只大鼠随机分为模型组8只和针刺组8只。针刺组予自制鼠袋内固定,针刺双侧"带脉"穴,深度约为4～5mm,接电针,疏密波,2Hz/15Hz,1.5mA,每次20min,隔日1次,干预8周。测量各组大鼠体质量、腹围、内脏脂肪及肝脏湿重,全自动生化仪检测大鼠血清总胆固醇(TC)、血清甘油三酯(TG),谷丙转氨酶(ALT)、谷草转氨酶(AST),CT扫描检测腹部内脏脂肪分布,肝组织匀浆测定丙二醇(MDA)、总超氧化物歧化酶(T-SOD)和谷胱甘肽过氧化物酶(GSH-PX),苏木精-伊红(HE)染色观察肝脏病理形态学改变。结果:针刺前及针刺4、8周时,模型组大鼠体质量、腹围、内脏脂肪质量、肝脏湿重较空白组明显升高(P<0.001,P<0.05)。针刺4、8周末,与模型组比较,针刺组大鼠的体质量、腹围明显下降(P<0.05),针刺组内脏脂肪、肝脏湿重较模型组明显降低(P<0.05)。针刺8周末,与空白组比较,模型组大鼠血清ALT、AST、TC、TG明显升高(P<0.001,P<0.01),与模型组比较,针刺组血清ALT、AST、TC、TG水平均显著下降(P<0.05),CT结果可见腹部内脏脂肪分布减少,肝脏病理形态学可见肝脏脂肪变性程度改善。与空白组比较,模型组大鼠肝组织MDA含量明显升高(P<0.001),T-SOD、GSH-PX活性降低(P<0.001);与模型组比较,针刺组MDA含量下降、T-SOD及GSH-PX活性升高(P<0.01)。结论:针刺可以减轻腹型肥胖型NAFLD大鼠体质量、腹围,减少腹部内脏脂肪堆积,改善肝脏脂质代谢,其机制可能与抑制氧化应激有关。
Objective To investigate the effect of acupuncture of"Daimai"(GB26)on abdominal fat accumulation,lipid metabolism and hepatic oxidative stress in abdominal obese non-alcoholic fatty liver disease(NAFLD)rats.Methods male SD rats were divided into 3 groups:normal diet(normal,n=8),high fat diet control(model)and acupuncture(n=8/group in the latter2 groups).The abdominal obese NAFLD model was established by feeding the rats with high fat diet for 12 weeks.EA(2 Hz/15 Hz,1.5 mA)was applied to bilateral GB26 for 20 min,once every other day for 8 weeks.Rats of the model group were also restrained for 20 min as those in the EA group.The body mass and abdominal circumference were measured weekly,the isolated adipose tissues around the bilateral kidney and epididymis and the fresh liver were weighed.The contents of serum cholesterol(TC),triglyceride(TG),alanine transaminase(ALT),glutamic oxaloacetic aminotransferase(AST)were detected by using an automatic biochemical analyzer.The abdominal visceral fat distribution was acquired by CT scanning.The oxidative stress indexes of the homogenated liver tissues,such as malondialdehyde(MDA)was assayed using sodium thiobarbital(TBA)method,and the activity of total superoxide dismutase(T-SOD)and glutathione peroxidase(GSH-PX)were assayed by using hydroxylamine method and colorimetric method respectively.The histopathological changes of the liver were observed after staining with hematoxylin-eosin(HE).Results Following modeling,the body mass and waist circumference,visceral fat weight of bilateral kidneys and testis(visceral fat weight),liver weight,serum ALT,AST,TG and TC and liver MDA contents,were significantly higher in the model group(P<0.001,P<0.05),while hepatic T-SOD and GSH-PX activity was considerably lower in the model group than those in the normal group(P<0.001).After acupuncture intervention,the levels of all the above-mentioned indexes(modeling induced both increase and decrease)were reversed relevant to the model group(P<0.05,P<0.01).The results of CT scanning showed that the fat accumulation area in the abdomen was 8.67 cm2,18.51 cm2 and 13.75 cm2 in the normal,model and acupuncture groups,respectively,presenting a decrease after acupuncture.H.E.staining displayed that the degree of hepatic steatosis(including vague hepatic lobule boundary,disordered arrangement of hepatic cord,hepatocellular swelling,diffuse fatty degeneration,unequal-sized lipid droplets in the hepatocytes,nucleus excursion and dissolution after modeling)was improved after acupuncture.Conclusion Acupuncture can reduce body weight and abdominal fat accumulation in abdominal obese NAFLD rats,which may be related to its effects in inhibiting oxidative stress(lowering MDA level and increasing the activity of T-SOD and GSH-PX)and improving hepatic lipid metabolism.
Objective To investigate the effect of acupuncture of"Daimai"(GB26)on abdominal fat accumulation,lipid metabolism and hepatic oxidative stress in abdominal obese non-alcoholic fatty liver disease(NAFLD)rats.Methods male SD rats were divided into 3 groups:normal diet(normal,n=8),high fat diet control(model)and acupuncture(n=8/group in the latter2 groups).The abdominal obese NAFLD model was established by feeding the rats with high fat diet for 12 weeks.EA(2 Hz/15 Hz,1.5 mA)was applied to bilateral GB26 for 20 min,once every other day for 8 weeks.Rats of the model group were also restrained for 20 min as those in the EA group.The body mass and abdominal circumference were measured weekly,the isolated adipose tissues around the bilateral kidney and epididymis and the fresh liver were weighed.The contents of serum cholesterol(TC),triglyceride(TG),alanine transaminase(ALT),glutamic oxaloacetic aminotransferase(AST)were detected by using an automatic biochemical analyzer.The abdominal visceral fat distribution was acquired by CT scanning.The oxidative stress indexes of the homogenated liver tissues,such as malondialdehyde(MDA)was assayed using sodium thiobarbital(TBA)method,and the activity of total superoxide dismutase(T-SOD)and glutathione peroxidase(GSH-PX)were assayed by using hydroxylamine method and colorimetric method respectively.The histopathological changes of the liver were observed after staining with hematoxylin-eosin(HE).Results Following modeling,the body mass and waist circumference,visceral fat weight of bilateral kidneys and testis(visceral fat weight),liver weight,serum ALT,AST,TG and TC and liver MDA contents,were significantly higher in the model group(P<0.001,P<0.05),while hepatic T-SOD and GSH-PX activity was considerably lower in the model group than those in the normal group(P<0.001).After acupuncture intervention,the levels of all the above-mentioned indexes(modeling induced both increase and decrease)were reversed relevant to the model group(P<0.05,P<0.01).The results of CT scanning showed that the fat accumulation area in the abdomen was 8.67 cm2,18.51 cm2 and 13.75 cm2 in the normal,model and acupuncture groups,respectively,presenting a decrease after acupuncture.H.E.staining displayed that the degree of hepatic steatosis(including vague hepatic lobule boundary,disordered arrangement of hepatic cord,hepatocellular swelling,diffuse fatty degeneration,unequal-sized lipid droplets in the hepatocytes,nucleus excursion and dissolution after modeling)was improved after acupuncture.Conclusion Acupuncture can reduce body weight and abdominal fat accumulation in abdominal obese NAFLD rats,which may be related to its effects in inhibiting oxidative stress(lowering MDA level and increasing the activity of T-SOD and GSH-PX)and improving hepatic lipid metabolism.
引文
[1]YOUNOSSI Z M,KOENIG A B,ABDELATIF D,et al.Global epidemiology of nonalcoholic fatty liver disease-Meta-analytic assessment of prevalence,incidence,and outcomes[J].Hepatology,2016,64(1):73-84.
[2]MAVROGIANNAKI A N,MIGDALIS I N.Nonalcoholic fatty liver disease,diabetes mellitus and cardiovascular disease:newer data[J].Int J Endocrinol,2013,2013:450639.
[3]ESTES C,ANSTEE Q M,ARIAS-LOSTE M T,et al.Modeling NAFLD disease burden in China,France,Germany,Italy,Japan,Spain,United Kingdom,and United States for the period2016-2030[J].J Hepatol,2018,69(4):896-904.
[4]WILLIAMS C D,STENGEL J,ASIKE M I,et al.Prevalence of nonalcoholic fatty liver disease and nonalcoholic steatohepatitis among a largely middle-aged population utilizing ultrasound and liver biopsy:aprospective study[J].Gastroenterology,2011,140(1):124-131.
[5]BELLENTANI S.The epidemiology of non-alcoholic fatty liver disease[J].Liver Int,2017,37(Suppl 1):81-84.
[6]HSIAO T J,CHEN J C,WANG J D.Insulin resistance and ferritin as major determinants of nonalcoholic fatty liver disease in apparently healthy obese patients[J].Int J Obes Relat Metab Disord,2004,28(1):167-172.
[7]YKI-JARVINEN H.Non-alcoholic fatty liver disease as a cause and a consequence of metabolic syndrome[J].Lancet Diabetes Endocrinol,2014,2(11):901-910.
[8]KOTRONEN A,YKI-JARVINEN H.Fatty liver:a novel component of the metabolic syndrome[J].Arterioscler Thromb Vasc Biol,2008,28(1):27-38.
[9]SMITS M M,IOANNOU G N,BOYKO E J,et al.Non-alcoholic fatty liver disease as an independent manifestation of the metabolic syndrome:results of a US national survey in three ethnic groups[J].J Gastroenterol Hepatol,2013,28(4):664-670.
[10]梁翠梅,胡慧,王朝歆,等.针刺治疗腹型肥胖随机对照临床试验[J].针刺研究,2016,41(2):159-162.
[11]李媛媛,胡慧,刘洋,等.针刺“治未病”防治代谢综合征的实验研究[J].中国中医基础医学杂志,2014,20(4):517-518.
[12]李媛媛,胡慧,梁翠梅,等.针刺“带脉”穴对代谢综合征大鼠体质量、血糖和脂代谢的影响[J].针刺研究,2014,39(3):202-206.
[13]王力,龚标,朱丽丽,等.电针对非酒精性脂肪肝大鼠血清及肝组织白介素-18的影响[J].针刺研究,2013,38(3):208-213.
[14]潘良,梁翠梅,李媛媛,等.以通调带脉为主干预代谢综合征腹型肥胖的思路探讨[J].上海针灸杂志,2017,36(12):1475-1479.
[15]PANG Q,ZHANG J Y,SONG S D,et al.Central obesity and nonalcoholic fatty liver disease risk after adjusting for body mass index[J].World J Gastroenterol,2015,21(5):1650-1662.
[16]DAY C P,JAMES O F.Steatohepatitis:a tale of two“hits”[J].Gastroenterology,1998,114(4):842-845.
[17]SCHMATZ R,PERREIRA L B,STEFANELLO N,et al.Effects of resveratrol on biomarkers of oxidative stress and on the activity of delta aminolevulinic acid dehydratase in liver and kidney of streptozotocin-induced diabetic rats[J].Biochimie,2012,94(2):374-383.
[18]BIRBEN E,SAHINER U M,SACKESEN C,et al.Oxidative stress and antioxidant defense[J].World Allergy Organ J,2012,5(1):9-19.
[19]SATAPATI S,KUCEJOVA B,DUARTE J A,et al.Mitochondrial metabolism mediates oxidative stress and inflammation in fatty liver[J].J Clin Invest,2015,125(12):4447-4462.
[20]VIDELA L A,RODRIGO R,ORELLANA M,et al.Oxidative stress-related parameters in the liver of non-alcoholic fatty liver disease patients[J].Clin Sci(Lond),2004,106(3):261-268.
[21]POLIMENI L,DEL B M,BARATTA F,et al.Oxidative stress:New insights on the association of non-alcoholic fatty liver disease and atherosclerosis[J].World J Hepatol,2015,7(10):1325-1336.
[22]YESILOVA Z,YAMAN H,OKTENLI C,et al.Systemic markers of lipid peroxidation and antioxidants in patients with nonalcoholic fatty liver disease[J].Am J Gastroenterol,2005,100(4):850-855.
[23]ASHRAF N U,SHEIKH T A.Endoplasmic reticulum stress and oxidative stress in the pathogenesis of non-alcoholic fatty liver disease[J].Free Radic Res,2015,49(12):1405-1418.
[24]沈凌宇,梁翠梅,杨文津,等.通调带脉法针刺治疗腹部肥胖型多囊卵巢综合征的随机对照研究[J].针刺研究,2018,43(4):255-259.
[25]李媛媛,胡慧,梁翠梅,等.针刺“带脉”穴对代谢综合征大鼠体质量、血糖和脂代谢的影响[J].针刺研究,2014,39(3):202-206.
[2]MAVROGIANNAKI A N,MIGDALIS I N.Nonalcoholic fatty liver disease,diabetes mellitus and cardiovascular disease:newer data[J].Int J Endocrinol,2013,2013:450639.
[3]ESTES C,ANSTEE Q M,ARIAS-LOSTE M T,et al.Modeling NAFLD disease burden in China,France,Germany,Italy,Japan,Spain,United Kingdom,and United States for the period2016-2030[J].J Hepatol,2018,69(4):896-904.
[4]WILLIAMS C D,STENGEL J,ASIKE M I,et al.Prevalence of nonalcoholic fatty liver disease and nonalcoholic steatohepatitis among a largely middle-aged population utilizing ultrasound and liver biopsy:aprospective study[J].Gastroenterology,2011,140(1):124-131.
[5]BELLENTANI S.The epidemiology of non-alcoholic fatty liver disease[J].Liver Int,2017,37(Suppl 1):81-84.
[6]HSIAO T J,CHEN J C,WANG J D.Insulin resistance and ferritin as major determinants of nonalcoholic fatty liver disease in apparently healthy obese patients[J].Int J Obes Relat Metab Disord,2004,28(1):167-172.
[7]YKI-JARVINEN H.Non-alcoholic fatty liver disease as a cause and a consequence of metabolic syndrome[J].Lancet Diabetes Endocrinol,2014,2(11):901-910.
[8]KOTRONEN A,YKI-JARVINEN H.Fatty liver:a novel component of the metabolic syndrome[J].Arterioscler Thromb Vasc Biol,2008,28(1):27-38.
[9]SMITS M M,IOANNOU G N,BOYKO E J,et al.Non-alcoholic fatty liver disease as an independent manifestation of the metabolic syndrome:results of a US national survey in three ethnic groups[J].J Gastroenterol Hepatol,2013,28(4):664-670.
[10]梁翠梅,胡慧,王朝歆,等.针刺治疗腹型肥胖随机对照临床试验[J].针刺研究,2016,41(2):159-162.
[11]李媛媛,胡慧,刘洋,等.针刺“治未病”防治代谢综合征的实验研究[J].中国中医基础医学杂志,2014,20(4):517-518.
[12]李媛媛,胡慧,梁翠梅,等.针刺“带脉”穴对代谢综合征大鼠体质量、血糖和脂代谢的影响[J].针刺研究,2014,39(3):202-206.
[13]王力,龚标,朱丽丽,等.电针对非酒精性脂肪肝大鼠血清及肝组织白介素-18的影响[J].针刺研究,2013,38(3):208-213.
[14]潘良,梁翠梅,李媛媛,等.以通调带脉为主干预代谢综合征腹型肥胖的思路探讨[J].上海针灸杂志,2017,36(12):1475-1479.
[15]PANG Q,ZHANG J Y,SONG S D,et al.Central obesity and nonalcoholic fatty liver disease risk after adjusting for body mass index[J].World J Gastroenterol,2015,21(5):1650-1662.
[16]DAY C P,JAMES O F.Steatohepatitis:a tale of two“hits”[J].Gastroenterology,1998,114(4):842-845.
[17]SCHMATZ R,PERREIRA L B,STEFANELLO N,et al.Effects of resveratrol on biomarkers of oxidative stress and on the activity of delta aminolevulinic acid dehydratase in liver and kidney of streptozotocin-induced diabetic rats[J].Biochimie,2012,94(2):374-383.
[18]BIRBEN E,SAHINER U M,SACKESEN C,et al.Oxidative stress and antioxidant defense[J].World Allergy Organ J,2012,5(1):9-19.
[19]SATAPATI S,KUCEJOVA B,DUARTE J A,et al.Mitochondrial metabolism mediates oxidative stress and inflammation in fatty liver[J].J Clin Invest,2015,125(12):4447-4462.
[20]VIDELA L A,RODRIGO R,ORELLANA M,et al.Oxidative stress-related parameters in the liver of non-alcoholic fatty liver disease patients[J].Clin Sci(Lond),2004,106(3):261-268.
[21]POLIMENI L,DEL B M,BARATTA F,et al.Oxidative stress:New insights on the association of non-alcoholic fatty liver disease and atherosclerosis[J].World J Hepatol,2015,7(10):1325-1336.
[22]YESILOVA Z,YAMAN H,OKTENLI C,et al.Systemic markers of lipid peroxidation and antioxidants in patients with nonalcoholic fatty liver disease[J].Am J Gastroenterol,2005,100(4):850-855.
[23]ASHRAF N U,SHEIKH T A.Endoplasmic reticulum stress and oxidative stress in the pathogenesis of non-alcoholic fatty liver disease[J].Free Radic Res,2015,49(12):1405-1418.
[24]沈凌宇,梁翠梅,杨文津,等.通调带脉法针刺治疗腹部肥胖型多囊卵巢综合征的随机对照研究[J].针刺研究,2018,43(4):255-259.
[25]李媛媛,胡慧,梁翠梅,等.针刺“带脉”穴对代谢综合征大鼠体质量、血糖和脂代谢的影响[J].针刺研究,2014,39(3):202-206.
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