基于MAPK途径探讨丹蛭降糖胶囊改善高脂血症大鼠肝损伤的机制
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摘要
探究丹蛭降糖胶囊对高脂血症大鼠肝脏损伤的保护作用及初步机制。采用高脂饲料喂养12周制备雄性SD大鼠高脂血症模型,模型成功后分为模型对照组、丹蛭降糖胶囊2个剂量组(500,1 000 mg·kg~(-1)·d-1),连续灌胃给药8周。分析血脂和肝代谢指标,HE和油红O染色观察肝脏的病理学变化,RT-PCR法分析细胞外信号调节蛋白激酶1/2(ERK1/2)、c-Jun氨基末端激酶(JNK)及p38丝裂原活化蛋白激酶(p38 MAPK)的表达,免疫组化和Western blot法分析p-ERK,p-JNK,p-p38和MCP-1的表达。结果显示,丹蛭降糖胶囊能明显降低大鼠体质量和血清TC,TG,ALT,AST水平,升高HDL-C水平,逆转高脂饮食诱导的肝损伤及脂质沉积,降低ERK1/2,JNK,p-38 MAPK mRNA水平,降低p-ERK,p-JNK,p-p38,MCP-1蛋白表达水平,且具有一定的剂量效应。因此,丹蛭降糖胶囊可通过抑制MAPK途径和炎症而改善高脂血症大鼠的肝损伤,并有一定的剂量效应。
This study aimed to investigate the protective effect and preliminary mechanism of Danzhi Jiangtang Capsules( DJC) on liver of hyperlipidemic rats. The hyperlipidemia models were successfully made by high-fat diet for 12 weeks in male SD rats,and then divided into model control group and DJC treatment groups( 500 and 1 000 mg·kg~(-1)·d-1) via gavage administration for additional 8 weeks.The levels of serum lipid and liver metabolism indices were detected; HE and oil red O staining were used to observe the pathological changes of liver. Expression levels of extracellular regulated protein kinase 1/2( ERK1/2),c-Jun N-terminal kinase( JNK),and p38 mitogen-activated protein kinase( p38 MAPK) were detected by real-time polymerase chain reaction( RT-PCR). Expression of MCP-1,phosphorylated ERK( p-ERK),phosphorylated JNK( p-JNK),and phosphorylated p38 MAPK( p-p38) were analyzed by immunohistochemistry and Western blot. The results showed that DJC decreased body weight and serum levels of total cholesterol( TC),triglyceride( TG),alanine aminotransferase( ALT),aspartate aminotransferase( AST),increased serum high-density lipoprotein cholesterol( HDL-C) level,ameliorate injury and lipid deposition in the liver induced by the high-fat diet,decreased mRNA expression of ERK1/2,JNK and p-38 MAPK as well as protein expression of p-ERK,p-JNK,p-p38,and MCP-1,somewhat showing a dose-dependent effect. Therefore,DJC has an obvious protective effect on liver of hyperlipidemic rats with certain dose-dependent effect,and the mechanism may be related with inhibiting MAPK pathways and inflammation.
This study aimed to investigate the protective effect and preliminary mechanism of Danzhi Jiangtang Capsules( DJC) on liver of hyperlipidemic rats. The hyperlipidemia models were successfully made by high-fat diet for 12 weeks in male SD rats,and then divided into model control group and DJC treatment groups( 500 and 1 000 mg·kg~(-1)·d-1) via gavage administration for additional 8 weeks.The levels of serum lipid and liver metabolism indices were detected; HE and oil red O staining were used to observe the pathological changes of liver. Expression levels of extracellular regulated protein kinase 1/2( ERK1/2),c-Jun N-terminal kinase( JNK),and p38 mitogen-activated protein kinase( p38 MAPK) were detected by real-time polymerase chain reaction( RT-PCR). Expression of MCP-1,phosphorylated ERK( p-ERK),phosphorylated JNK( p-JNK),and phosphorylated p38 MAPK( p-p38) were analyzed by immunohistochemistry and Western blot. The results showed that DJC decreased body weight and serum levels of total cholesterol( TC),triglyceride( TG),alanine aminotransferase( ALT),aspartate aminotransferase( AST),increased serum high-density lipoprotein cholesterol( HDL-C) level,ameliorate injury and lipid deposition in the liver induced by the high-fat diet,decreased mRNA expression of ERK1/2,JNK and p-38 MAPK as well as protein expression of p-ERK,p-JNK,p-p38,and MCP-1,somewhat showing a dose-dependent effect. Therefore,DJC has an obvious protective effect on liver of hyperlipidemic rats with certain dose-dependent effect,and the mechanism may be related with inhibiting MAPK pathways and inflammation.
引文
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[26]庄星星,杨晓旭,魏良兵,等. UPLC指纹图谱法考察丹蛭降糖胶囊的不同制备工艺[J].中成药,2015,37(1):207.
[27] Lu L,Qin Y,Chen C,et al. Beneficial effects exerted by paeonol in the management of atherosclerosis[J].Oxid Med Cell Longev,2018,2018:1098617.
[28] Peng L,Pan X,Yin G. Natural hirudin increases rat flap viability by anti-inflammation via PARs/p38/NF-κB pathway[J]. Biomed Res Int,2015,2015:597264.
[29]龚杰,丁岩,干仲元,等.泽泻提取物对大鼠非酒精性脂肪肝的治疗作用[J].中国比较医学杂志,2018,28(7):68.
[30]吴晶魁,杨乔.中药水蛭对高脂血症大鼠脂质代谢及肝脏的影响[J].中国中药杂志,2018,43(4):794.
[2] Younossi Z M. Non-alcoholic fatty liver disease-a global public[J]. J Hepatol,2018,doi:10. 1016/j.jhep.2018. 10. 033.
[3] Finck B N. Targeting metabolism,insulin resistance,and diabetes to treat nonalcoholic steatohepatitis[J]. Diabetes,2018,67(12):2485.
[4] Mazhar K. The future of nonalcoholic fatty liver disease treatment[J]. Med Clin North Am,2019,103(1):57.
[5] Polyzos S A,Kountouras J,Mantzoros C S. Obesity and nonalcoholic fatty liver disease:from pathophysiology to therapeutics[J].Metabolism,2019,92:82.
[6]赵进东,丁雷,鲍陶陶,等.丹蛭降糖胶囊对2型糖尿病患者血脂作用有效性的系统评价[J].中华中医药学刊,2015,33(3):570.
[7]李中南,方朝晖,熊园园,等.丹蛭降糖胶囊对糖尿病大鼠肝脏PI3K/AKT信号通路的影响[J].上海中医药杂志,2015,49(9):67.
[8]李中南,马超,程丽华,等.丹蛭降糖胶囊对糖尿病大鼠肝组织HIF-1α的干预及肝脏病理的影响[J].中成药,2017,39(4):831.
[9]陈明卫,夏同佳,方朝晖,等.丹蛭降糖胶囊改善高脂饮食诱导的肥胖大鼠骨骼肌胰岛素抵抗机制的初步研究[J].中华临床医师杂志:电子版,2014,8(9):1697.
[10] Mintziori G,Polyzos S A. Emerging and future therapies for nonalcoholic steatohepatitis in adults[J]. Expert Opin Pharmacother,2016,17:1937.
[11]周珂,刘忠第,谭勇,等.高尿酸血症和非酒精性脂肪肝异病同治规律及分子机制预测[J].中国实验方剂学杂志,2017,23(1):206.
[12] Zhang N,Lu Y,Shen X,et al. Fenofibrate treatment attenuated chronic endoplasmic reticulum stress in the liver of nonalcoholic fatty liver disease mice[J]. Pharmacology,2015,95(3/4):173.
[13] Chen M,Deng D,Fang Z,et al,Fenofibrate increases serum vaspin by upregulating its expression in adipose tissue[J]. Endocrine,2014,45(3):409.
[14]龚杰,丁岩,干仲元,等.泽泻提取物对大鼠非酒精性脂肪肝的治疗作用[J].中国比较医学杂志,2018,28(7):68.
[15]段卫娜,张振凌,孔莹莹,等.地黄不同炮制品组成的增液汤降低糖尿病大鼠血糖血脂作用的对比研究[J].中国实验方剂学杂志,2013,19(6):187.
[16]潘雪,马端鑫,李燕,等.水蛭药理作用的研究进展[J].中国民族民间医药,2015,24(14):24.
[17]吴斌,张一,陈勇,等.丹蛭降糖胶囊治疗肥胖引起的慢性肾病的机制研究[J].中国药理学通报,2018,34(8):1163.
[18] Lu Y,Chen Y,Li R,et al. Protective effects of Danzhi jiangtang capsule on vascular endothelial damages induced by high-fat diet and palmitic acid[J].Biomed Pharmacother,2018,107:1631.
[19] You Z,Shao-Peng L,Jing D,et al. Advancements in MAPK signaling pathways and MAPK-targeted therapies for ameloblastoma:a review[J]. J Oral Pathol Med,2018,doi:10. 1111/jop.12807.
[20] Nandipati K C,Subramanian S,Agrawal D K. Protein kinases:mechanisms and downstream targets in inflammation-mediated obesity and insulin resistance[J]. Mol Cell Biochem,2017,426(1/2):27.
[21]杨晓春,鲍陶陶,储全根.基于p38MAPK通路探讨丹蛭降糖胶囊对2型糖尿病模型大鼠血管病变的影响[J].中国实验方剂学杂志,2016,22(3):116.
[22] Deng Y,Tang K,Chen R,et al. Effects of Shugan-Jianpi recipe on the expression of the p38 MAPK/NF-κB signaling pathway in the hepatocytes of NAFLD rats[J]. Medicines,2018,5(3):E106.
[23] Sun H,Wang X,Chen J,et al. Melatonin improves non-alcoholic fatty liver disease via MAPK-JNK/P38 signaling in high-fatdiet-induced obese mice[J]. Lipids Health Dis,2016,15(1):202.
[24] Lawan A,Bennett A M. Mitogen-activated protein kinase regulation in hepatic metabolism[J]. Trends Endocrinol Metab,2017,28(12):868.
[25] Knebel B,Lehr S,Hartwig S,et al. Phosphorylation of sterol regulatory element-binding protein(SREBP)-1c by p38 kinases,ERK and JNK influences lipid metabolism and the secretome of human liver cell line Hep G2[J]. Arch Physiol Biochem,2014,120(5):216.
[26]庄星星,杨晓旭,魏良兵,等. UPLC指纹图谱法考察丹蛭降糖胶囊的不同制备工艺[J].中成药,2015,37(1):207.
[27] Lu L,Qin Y,Chen C,et al. Beneficial effects exerted by paeonol in the management of atherosclerosis[J].Oxid Med Cell Longev,2018,2018:1098617.
[28] Peng L,Pan X,Yin G. Natural hirudin increases rat flap viability by anti-inflammation via PARs/p38/NF-κB pathway[J]. Biomed Res Int,2015,2015:597264.
[29]龚杰,丁岩,干仲元,等.泽泻提取物对大鼠非酒精性脂肪肝的治疗作用[J].中国比较医学杂志,2018,28(7):68.
[30]吴晶魁,杨乔.中药水蛭对高脂血症大鼠脂质代谢及肝脏的影响[J].中国中药杂志,2018,43(4):794.