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首页|期刊导航|北京中医药大学学报|葛花解酒涤脂汤对酒精性肝病小鼠肝脏组织能量代谢的影响

葛花解酒涤脂汤对酒精性肝病小鼠肝脏组织能量代谢的影响OA北大核心CSTPCD

Effect of Gehua Jiejiu Dizhi Decoction on energy metabolism in liver tissue of alcoholic liver disease mice

中文摘要英文摘要

目的 检测酒精性肝病(ALD)小鼠肝脏组织能量代谢相关指标水平,并探讨葛花解酒涤脂汤的干预作用.方法 40 只雄性C57BL/6J小鼠任意分为正常对照组、模型组、葛花解酒涤脂汤高剂量组(4.94 g/kg)、葛花解酒涤脂汤低剂量组(2.47 g/kg)和白藜芦醇组(0.40 g/kg),每组 8 只.除正常对照组外,其他各组小鼠先予Lieber-DeCarli对照液体饲料喂养5 d,随后予Lieber-DeCarli酒精液体饲料喂养10 d,第16 天灌胃31.5%乙醇溶液1 次,以制备ALD小鼠模型.造模结束后第2 天起,各干预组灌胃相应药物,每日1 次,连续9 d.苏木素-伊红染色和油红O染色观察肝脏组织脂肪变性情况;分光光度法检测肝脏组织Na+K+-腺苷三磷酸(ATP)酶、Ca2+Mg2+-ATP 酶活性,琥珀酸脱氢酶(SDH)、肝糖原含量;反相高效液相色谱法检测肝脏组织ATP、腺苷二磷酸(ADP)和腺苷一磷酸(AMP)含量,以及AMP/ATP值、总腺苷酸池(TAN)含量、能荷值(EC);实时荧光PCR法检测肝脏组织沉默信息调节因子1(SIRT1)、腺苷酸活化蛋白激酶(AMPK)α2 mRNA表达;蛋白质印迹法检测肝脏组织SIRT1、AMPKα2、AMPKβ1 蛋白表达.结果 与正常对照组比较,模型组小鼠肝脏组织脂肪变性病变明显,肝脏组织Ca2+Mg2+-ATP 酶活性、SDH含量降低,肝糖原含量升高,EC、AMP/ATP 值降低,ATP、ADP、AMP和TAN含量升高,SIRT1、AMPKα2 mRNA表达降低,AMPKβ1 蛋白表达升高(均P<0.05).与模型组比较,葛花解酒涤脂汤高、低剂量组肝脏组织脂肪变性明显减轻,肝脏组织Na+K+-ATP酶活性降低,EC及SIRT1、AMPKα2 mRNA表达升高(均P<0.05);葛花解酒涤脂汤低剂量组Ca2+Mg2+-ATP酶活性、SDH及ATP含量升高(均P<0.05);葛花解酒涤脂汤高剂量组AMP/ATP值升高(P<0.05);葛花解酒涤脂汤高、低剂量组和白藜芦醇组肝脏组织SIRT1 蛋白表达升高(均P<0.05);葛花解酒涤脂汤低剂量组和白藜芦醇组AMPKα2 蛋白表达升高(均P<0.05).与葛花解酒涤脂汤高剂量组比较,葛花解酒涤脂汤低剂量组、白藜芦醇组肝脏组织ATP、TAN含量和EC升高,AMP/ATP值降低(均P<0.05);葛花解酒涤脂汤低剂量组AMPKα2 mRNA表达降低(P<0.05);白藜芦醇组SIRT1、AMPKα2 蛋白表达升高(均P<0.05).与葛花解酒涤脂汤低剂量组比较,白藜芦醇组AMPKβ1 蛋白表达降低(P<0.05).结论 慢性酒精摄入引起的能量代谢改变可能与小鼠ALD发生有关,葛花解酒涤脂汤干预可以改善ALD小鼠肝脏能量代谢异常.

Objective To detect the level of related indexes of energy metabolism in liver tissue of alcoholic liver disease(ALD)mice,and to explore the intervention effect of Gehua Jiejiu Dizhi Decoction.Methods Forty male C57BL/6J mice were randomly divided into the normal control group,the model group,the Gehua Jiejiu Dizhi Decoction high-dose group(4.94 g/kg),the Gehua Jiejiu Dizhi Decoction low-dose group(2.47 g/kg),and the resveratrol group(0.40 g/kg),with 8 mice in each group.Except the normal control group,the mice in other groups were fed with Lieber-DeCarli control liquid diet for five days,followed by Lieber-DeCarli alcohol liquid diet for ten days,and on the 16th day,the mice were given 31.5%alcohol solution through gavage to establish the ALD model.From the second day after modeling,the rats in the intervention groups were given the corresponding drugs by gavage once a day for nine consecutive days.Hematoxylin and eosin staining and oil red O staining were used to observe the liver steatosis in liver tissue.The activities of Na+K+-ATPase and Ca2+Mg2+-ATPase,and the contents of succinate dehydrogenase(SDH)and hepatic glycogen in liver tissue were detected using spectrophotometry.The contents of ATP,ADP,AMP,the AMP/ATP value,total adenosine pool(TAN)content,and energy charge(EC)in liver tissue were detected by reversed phase high performance liquid chromatography method.The mRNA expressions of NAD dependent deacetylase Sirtuin-1(SIRT1)and AMP-activated protein kinase(AMPK)α2 in liver tissue were detected by real-time PCR.The protein expressions of SIRT1,AMPKα2,and AMPKβ1 in liver tissue were detected by Western blotting.Results Compared with the normal control group,the model group mice showed significant hepatic steatosis,significantly decreased Ca2+Mg2+-ATPase activity and SDH content in liver tissue,significantly increased hepatic glycogen content,significantly decreased EC and AMP/ATP value,significantly increased ATP,ADP,AMP,and TAN content,significantly decreased mRNA expressions of SIRT1 and AMPKα2,significantly increased protein expression of AMPKβ1(P<0.05).Compared with the model group,the Gehua Jiejiu Dizhi Decoction high-and low-dose groups significantly reduced liver tissue steatosis,and the activity of Na+K+-ATPase in liver tissue was significantly reduced,the EC and the mRNA expressions of SIRT1 and AMPKα2 were increased(P<0.05);the activity of Ca2+Mg2+-ATPase,SDH and ATP contents were increased in the Gehua Jiejiu Dizhi Decoction low-dose group(P<0.05);the AMP/ATP value was increased in the Gehua Jiejiu Dizhi Decoction high-dose group(P<0.05);and the protein expression of SIRT1 was increased in the the Gehua Jiejiu Dizhi Decoction high-and low-dose groups and the resveratrol group(P<0.05);the protein expression of AMPKα2 in the Gehua Jiejiu Dizhi Decoction low-dose group and the resveratrol group was increased(P<0.05).Compared with the Gehua Jiejiu Dizhi Decoction high-dose group,the Gehua Jiejiu Dizhi Decoction low-dose group and the resveratrol group showed a significant increase in ATP,TAN contents,and EC in liver tissue,while the AMP/ATP value decreased(P<0.05);mRNA expressionin of AMPKα2 in the Gehua Jiejiu Dizhi Decoction low-dose group was decreased(P<0.05);and the protein expressions of SIRT1 and AMPKα2 in the resveratrol group were increased(P<0.05).Compared with the Gehua Jiejiu Dizhi Decoction low-dose group,the protein expression of AMPKβ1 was decreased in the resveratrol group(P<0.05).Conclusion The changes of energy metabolism caused by chronic alcohol intake may be related to the occurrence of ALD,and the intervention of Gehua Jiejiu Dizhi Decoction can improve the abnormal energy metabolism in the liver of ALD mice.

刘蕊;易旭;吴雪莉;王硕石;何典城;游绍伟

贵州中医药大学第二附属医院临床医学实验室 贵阳 550002贵州中医药大学第二附属医院消化内科

中医学

酒精性肝病葛花解酒涤脂汤能量代谢沉默信息调节因子1腺苷酸活化蛋白激酶小鼠

alcoholic liver diseaseGehua Jiejiu Dizhi Decoctionenergy metabolismSIRT1AMPKmice

《北京中医药大学学报》 2024 (005)

662-671 / 10

国家自然科学基金地区基金项目(No.81660752);贵州省基础研究计划项目(黔科合基础-ZK[2023]一般410) National Natural Science Foundation of China(No.81660752)

10.3969/j.issn.1006-2157.2024.05.010

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