Objective To investigate the effect of the liver-regulating and spleen-strengthening method on lipid metabolism and expression of microtubule-associated protein light chain 3 B (LC3B) in a rat model of nonalcoholic fatty liver disease (NAFLD) , as well as the mechanism of this method in the treatment of NAFLD.Methods A total of 30 clean male Sprague-Dawley rats were randomly divided into control group, model group, and liver-regulating and spleen-strengthening group, with 10 rats in each group.The rats in the model group and the liver-regulating and spleen-strengthening group were given high-fat diet for 12 weeks.The rats in the liver-regulating and spleen-strengthening group were given liver-regulating and spleen-strengthening prescription 2 ml·100 g-1·d (five times the dose for adults) by gavage, and those in the control group were given normal diet and an equal volume of normal saline by gavage.The rats were sacrificed at the end of week 12, and blood and liver samples were collected to measure the serum levels of alanine aminotransferase (ALT) , aspartate aminotransferase (AST) , triglyceride (TG) , total cholesterol (TC) , high-density lipoprotein cholesterol (HDL-C) , and low-density lipoprotein cholesterol (LDL-C) .HE staining was performed to observe hepatic steatosis, and immunohistochemistry was used to measure the change in the expression of LC3B in liver tissue.A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups; the chi-square test was used for comparison of categorical data between groups; the Kruskal-Wallis H test was used for comparison of ranked data between multiple groups, and the Mann-Whitney U test was used for further comparison between two groups.Results HE staining showed that there were significant differences in the degree of hepatocyte fatty degeneration and score of inflammatory activity between the model group and the control group (both P < 0.001) ; compared with the model group, the liver-regulating and spleen-strengthening group had significant reductions in the degree of hepatocyte fatty degeneration and score of inflammatory activity (both P < 0.01) .Compared with the control group, the model group had significant increases in the serum levels of ALT, AST, TG, TC, HDL-C, and LDL-C (all P < 0.001) ; compared with the model group, the liver-regulating and spleen-strengthening group had significant reductions in the serum levels of ALT, AST, TG, TC, and LDL-C (all P < 0.05) .Compared with the control group, the model group had a slight increase in the expression of LC3B in liver tissue (χ2= 1.250, P > 0.05) ; compared with the model group, the liver-regulating and spleen-strengthening group had a significant increase in the expression of LC3B in liver tissue (χ2= 5.051, P < 0.05) .Conclusion The liver-regulating and spleen-strengthening method can improve liver function and blood lipids in experimental male Sprague-Dawley rats with NAFLD, possibly by increasing the level of autophagy in liver tissue.
[1]LEUNG TM, NIETO N.CYP2E1 and oxidant stress in alcoholic and non-alcoholic fatty liver disease[J].J Hepatol, 2013, 58 (2) :395-398.
|
[2]LEAMY AK, EGNATCHIK RA, YOUNG JD.Molecular mechanisms and the role of saturated fatty acids in the progression of nonalcoholic fatty liver disease[J].Prog Lipid Res, 2013, 52 (1) :165-174.
|
[3]ZHOU T, ZHANG SS, YU Q.Clinical cohort study on non-alcoholic fatty liver disease by regulating liver and spleen[J].Beijing J Tradit Chin Med, 2013, 32 (6) :403-405. (in Chinese) 周滔, 张声生, 郁强.调肝理脾法治疗非酒精性脂肪肝的临床队列研究[J].北京中医药, 2013, 32 (6) :403-405.
|
[4]ZHOU T, ZHANG SS, CHEN J.Analysis of 120 cases of nonalcoholic fatty liver unit syndrome and the clinical effect of Jianpi Qinggan recipe[J].Chin J Integr Tradit West Med Liver Dis, 2009, 19 (4) :209-210. (in Chinese) 周滔, 张声生, 陈誩.120例非酒精性脂肪肝单元证特点分析与健脾清肝方临床疗效[J].中西医结合肝病杂志, 2009, 19 (4) :209-210.
|
[5]SINGH R, KAUSHIK S, WANG Y, et al.Autophagy regulates lipid m etabolism[J].Nature, 2009, 458 (7242) :1131-1135.
|
[6]WU X, ZHANG L, GURLEY E, et al.Prevention of free fatty acid-induced hepatic lipotoxicity by 18beta-glycytthetinic acid through lysosomal and mitochondrial pathways[J].Hepatology, 2008, 47 (6) :1905-1915.
|
[7]DIEHL A, GOODMAN Z, ISHAK K.Alcohollike liver disease in nonalcoholics.A clinical and histologic comparison with alcoholinduced liver injury[J].Gastroenterology, 1998, 95 (4) :1056-1062.
|
[8]WANG TL, LIU X, ZHOU YP, et al.A semiquantitative scoring system for assessmeng of hepatic inflammation and fibrosis in chronic viral hepatitis[J].Chin J Hepatol, 1998, 6 (4) :195-197. (in Chinese) 王泰龄, 刘霞, 周元平, 等.慢性肝炎炎症活动度及纤维化程度计分方案[J].中华肝脏病杂志, 1998, 6 (4) :195-197.
|
[9]CHOI J, JUNG W, KOO JS.Expression of autophagy-related markers Beclin1, light chain 3A, light chain 3B and P62 according to the molecular subtype of breast cancer[J].Histopsthology, 2013, 62 (2) :275-286.
|
[10]KIM YC, GUAN KL.m TOR:a pharmacologic target for autophagy regulation[J].J Clin Invest, 2015, 125 (1) :25-32.
|
[11]WANG EY, BIALA AK, GORDON JW, et al.Autophagy in the heart:too much of a good thing?[J].J Cardiovasc Pharmacol, 2012, 60 (2) :110-117.
|
[12]MIZUSHIMA N, KOMATSU M.Autophagy:renovation of cells and tissues[J].Cell, 2011, 147 (4) :728-741.
|
[13]ROBINET P, RITCHEY B, SMITH JD.Physiological difference in autophagic flux in macrophages from 2 mouse strains regulates cholesterol ester metabolism[J].Arterioscler Thromb Vasc Biol, 2013, 33 (5) :903-910.
|
[14]LIU K, CZAJA MJ.Regulation of lipid stores and metabolism by lipophagy[J].Cell Death Differ, 2013, 20 (1) :3-11.
|
[15]ZHANG J, PENG XM, LIU ZQ.Pathogenic mechanism of endoplasmic reticulum stress in nonalcoholic fatty liver disease[J].Chin J Immunol, 2017, 33 (7) :1112-1114. (in Chinese) 张娟, 彭晓蔓, 刘朝奇.内质网应激在非酒精性脂肪性肝炎中的致病机制[J].中国免疫学杂志, 2017, 33 (7) :1112-1114.
|
[16]YANG L, LI P, FU S, et al.Defective hepatic antophagy in obesity promotes ER stress and causes insulin resistance[J].Cell Metab, 2010, 11 (6) :467-478.
|
[17]PAPACKOVA Z, DANKOVA H, PALENICKOVA E, et al.Effect of short-and long-term high-fat feeding on autophagy flux and lysosomal activity in rat liver[J].Physiol Res, 2012, 61 (Suppl2) :s67-s76.
|
[18]MARTINEZ-VICENTE M, TALLOCZY Z, WONG E, et al.Cargo recognition failure is responsible for inefficient autophagy in Huntington’s disease[J].Nat Neurosci, 2010, 13 (5) :567-576.
|
[19]CZAJA MJ.Autophagy in health and disease.2.Regulation of lipid metabolism and storage by autophagy:pathophysiological implications[J].Am J Physiol Cell Physiol, 2010, 298 (5) :c973-c978.
|
[20]WANG KY, LI XL, CAI YQ, et al.Research progress of the regulation function of autophagy in drug-induced hepatotoxicity[J].Chin J Clin Pharmacol Ther, 2017, 22 (1) :100-103. (in Chinese) 王楷扬, 李小丽, 蔡永青.自噬对药物肝毒性调控作用的研究进展[J].中国临床药理学与治疗学, 2017, 22 (1) :100-103.
|
[21]YAN R, NIU CY, YU L, et al.Interaction between autophagy and lipid metabolism in cell models of nonalcoholic fatty liver disease[J].J Clin Hepatol, 2017, 33 (10) :1981-1986. (in Chinese) .闫蓉, 牛春燕, 于璐, 等.非酒精性脂肪性肝病细胞模型中自噬与脂质代谢的相互调节[J].临床肝胆病杂志, 2017, 33 (10) :1981-1986.
|
[22]SHIBATA M, YOSHIMURA K, FURUYA N, et al.The MAP1-LC3conjugation system is involved in lipid droplet formation[J].Biochem Biophys Res Commun, 2009, 382 (2) :419-423.
|
[23]MARTINET W, SCHRIJVERS DM, TIMMERMANS JP, et al.Immunohisto-chemical analysis of macroautophagy:recommendations and limitations[J].Autophagy, 2013, 9 (3) :386-402.
|
[24]LIANG C.Negative regulation of autophagy[J].Cell Death Differ, 2010, 17 (12) :1807-1815.
|
[25]ZHU D, SUN TT, CHEN LY.Treatment of nonalcoholic fatty liver disease in Jingui Yaolue[J].J Changchun Univ Chin Med, 2016, 32 (2) :423-429. (in Chinese) 朱丹, 孙婷婷, 陈兰羽.《金匮要略》诸方治疗非酒精性脂肪性肝病进展[J].长春中医药大学学报, 2016, 32 (2) :423-429.
|
[26]HUANG LQ, HUANG GH, JI YX, et al.The correlation between autophagy and the"Yin and Yang"and"Qi"of traditional Chinese Medicine[J].Liaoning J Tradit Chin Med, 2014, 41 (6) :1147-1149. (in Chinese) 黄丽琼, 黄贵华, 纪云西, 等.细胞自噬与中医“阴阳”及“气”理论相关性探讨[J].辽宁中医杂志, 2014, 41 (6) :1147-1149.
|
[27]LI YX, AO HQ.Explore the relevance of autophagy and liver dispersion function from the microscopic point of view[J].Liaoning J Tradit Chin Med, 2016, 18 (2) :45-47. (in Chinese) 黎颖贤, 敖海清.从微观角度探究细胞自噬与肝主疏泄功能相关性[J].辽宁中医药大学学报, 2016, 18 (2) :45-47.
|
[28]JIANG WX, WANG YC, CHANG C.Discussion on Chinese medicine in regulation of autophagy[J].J Tradit Chin Med, 2017, 58 (18) :1566-1568. (in Chinese) 姜文秀, 王英超, 常诚.中医药调节细胞自噬刍议[J].中医杂志, 2017, 58 (18) :1566-1568.
|