复方茵丹汤对急性肝内胆汁淤积大鼠肝组织法尼醇受体及多耐药相关蛋白3表达的影响

隋京利; 孙凤霞; 李晓玲; 张秋香

doi:10.3969/j.issn.1001-5256.2018.09.026

复方茵丹汤对急性肝内胆汁淤积大鼠肝组织法尼醇受体及多耐药相关蛋白3表达的影响

DOI: 10.3969/j.issn.1001-5256.2018.09.026

1. 北京中医医院顺义医院感染科
2. 首都医科大学附属北京中医医院感染科

基金项目:

国家自然基金面上项目(81573897)；顺义区科技三项费项目(SYKW201602)；

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中图分类号: R285.5
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出版历程
- 收稿日期: 2018-03-15
- 出版日期: 2018-09-20

Effect of Compound Yindan Decoction on the expression of farnesoid X receptor and multidrug resistance-associated protein 3 in rats with acute intrahepatic cholestasis

Department of Infectious Diseases, Beijing Hospital of Traditional Chinese Medicine Shunyi Branch

Research funding:

摘要

摘要:
目的研究复方茵丹汤对α-萘异硫氰酸酯（ANIT）诱导的大鼠急性肝内胆汁淤积的作用机制。方法将108只雄性SD大鼠随机分为正常对照组,模型组,中药复方茵丹汤高、中、低剂量组和阳性药对照组,每组18只,依据取材时间不同又将各组随机分为24、48、72 h 3个亚组,每组6只。模型组、中药组、阳性药对照组大鼠一次性给予2%ANIT按100 mg·kg-1·d-1灌胃造模,正常对照组予等容积色拉油灌胃。造模2 h后,中药高、中、低剂量组分别给予生药含量为24.48 g·kg-1·d-1、12.24 g·kg-1·d-1和6.12 g·kg-1·d-1的复方茵丹汤中药煎剂l ml/100 g体质量灌胃,模型组和正常对照组给予等容积生理盐水灌胃,阳性药对照组给予67.5 mg·kg-1·d-1熊去氧胆酸（UDCA）水溶液l ml/100 g体质量灌胃,均1次/d。造模后分别于24、...
- 胆汁淤积,肝内 /
- 法尼醇 /
- 多耐药相关蛋白3 /
- 大鼠,Sprague-Dawley
Abstract:
Objective To investigate the mechanism of action of Compound Yindan Decoction on acute intrahepatic cholestasis induced byα-naphthyl isothiocyanate (ANIT) in rats. Methods A total of 108 male Sprague-Dawley rats were randomly divided into normal control group, model group, high-, middle-, and low-dose traditional Chinese medicine (TCM) groups, and positive control group, with 18 rats in each group. According to the time of sampling, each group was divided into 24-hour, 48-hour, and 72-hour subgroups, with 6 rats in each subgroup. The model group, the TCM groups, and the positive control group were given a single dose of 2% ANIT (100 mg/kg/d) by gavage to establish the model, and the normal control group was given an equal volume of salad oil by gavage. At 2 hours after modeling, the high-, middle-, and low-dose TCM groups were given Compound Yindan Decoction 1 ml/100 g body weight once a day by gavage at a dose of 24. 48 g·kg-1·d-1, 12. 24 g·kg-1·d-1, and 6. 12 g·kg-1·d-1; the model group and the normal control group were given an equal volume of normal saline once a day by gavage; the positive control group was given ursodeoxycholic acid (UDCA) solution 1 ml/100 g body weight once a day by gavage at a dose of 67. 5 mg·kg-1·d-1. The rats were sacrificed at 24, 48, or 72 hours after modeling, and RT-q-PCR, Western blot, and immunohistochemistry were used to measure the expression of farnesoid X receptor (FXR) and multidrug resistance-associated protein 3 (MRP3) in liver tissue. A one-way analysis of variance was used for comparison between multiple groups, and LSD-t test was used for further comparison between two groups. Results According to the results of RT-q-PCR, compared with the normal control group at each time phase, the model group had a significant reduction in the mRNA expression of FXR (all P<0. 05) ; compared with the model group at each time phase, all the TCM groups and the UDCA group had a significant increase in the mRNA expression of FXR (all P < 0. 05) ; the high-dose TCM group had significantly higher mRNA expression of FXR than the middle-and low-dose TCM groups at each time phase (all P < 0. 05) . Compared with the normal control group at each time phase, the model group had a significant increase in the mRNA expression of MRP3 (all P < 0. 05) ; compared with the model group at each time phase, the UDCA group and all the TCM groups except the 24-hour low-dose TCM subgroup had a significant increase in the mRNA expression of MRP3 (all P <0. 05) ; the 48-and 72-hour high-dose TCM subgroups had significantly higher mRNA expression of MRP3 than the middle-and low-dose TCM groups (all P < 0. 05) . According to the results of immunohistochemistry, compared with the normal control group at each time phase, the model group had a significant reduction in the protein expression of FXR (all P < 0. 05) ; compared with the model group, the 24-hour high-, middle-, and low-dose TCM subgroups and the UDCA group at each time phase had a significant increase in the protein expression of FXR (all P < 0. 05) ; the high-dose TCM group had significantly higher protein expression of FXR than the middle-and low-dose TCM groups at each time phase (all P < 0. 05) . Compared with the normal control group at each time phase, the model group had a significant increase in the protein expression of MRP3 (all P < 0. 05) ; compared with the model group at each time phase, the UDCA group and all the TCM groups except the 24-hour low-dose TCM subgroup had a significant increase in the protein expression of MRP3 (all P <0. 05) ; the high-dose TCM group had significantly higher protein expression of MRP3 than the low-dose TCM group at each time phase and the 72-hour middle-dose TCM subgroup (all P < 0. 05) . According to the results of Western blot, compared with the normal control group at each time phase, the model group had a significant reduction in the protein expression of FXR (all P < 0. 05) ; compared with the model group at each time phase, the high-, middle-, and low-dose TCM groups and the UDCA group had a significant increase in the protein expression of FXR (all P < 0. 05) ; the high-dose TCM group had significantly higher protein expression of FXR than the middle-and low-dose TCM groups at each time phase (all P < 0. 05) . Compared with the normal control group at each time phase, the model group had a significant increase in the protein expression of MRP3 (all P < 0. 05) ; compared with the model group, the 48-and 72-hour middle-and high-dose TCM subgroups and the UDCA group had a significant increase in the protein expression of MRP3 (all P < 0. 05) ; at 48 and72 hours, the high-dose TCM group had significantly higher protein expression of MRP3 than the middle-and low-dose TCM groups (all P < 0. 05) . Conclusion Compound Yindan Decoction has a therapeutic effect on rats with acute intrahepatic cholestasis induced by ANIT through gavage, possibly by upregulating the mRNA and protein expression of FXR and MRP3.
- cholestasis, intrahepatic /
- farnesol /
- multidrug resistance-associated protein 3 /
- rats, Sprague-Dawley

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参考文献(13)

[1]KHALAF R, PHEN C, KARJOO S, et al.Cholestasis beyond the neonatal and infancy periods[J].Pediatr Gastroenterol Hepatol Nutr, 2016, 19 (1) :1-11.

[2]SUN FX, SUI JL, LI P, et al.Biochemical parameters in diagnosis of cholestatic liver disease[J].J Clin Hepatol, 2016, 32 (8) :1488-1490. (in Chinese) 孙凤霞, 隋京利, 李攀, 等.胆汁淤积性肝病诊断相关生化指标的探讨[J].临床肝胆病杂志, 2016, 32 (8) :1488-1490.

[3]Expert Committee on the 2015 update of expert consensus on the diagnosis and treatment of cholestatic liver disease.Expert consensus on the diagnosis and treatment of cholestatic liver disease:An update in 2015[J].J Clin Hepatol, 2015, 31 (10) :1563-1574. (in Chinese) 胆汁淤积性肝病诊断治疗专家共识2015年更新专家委员会.胆汁淤积性肝病诊断治疗专家共识:2015年更新[J].临床肝胆病杂志, 2015, 31 (10) :1563-1574.

[4]WANG JM, SUN FX.Clinical effect evaluation of compound Yindan Decoction in treating acute cholestatic hepatitis[D].Beijing:Beijing Univ Chin Med, 2013. (in Chinese) 王建美, 孙凤霞.复方茵丹汤治疗急性淤胆型肝炎的临床疗效评价[D].北京:北京中医药大学, 2013.

[5]YUAN ZQ, LI KW.Role of farnesoid X receptor in cholestasis[J].J Dig Dis, 2016, 17 (8) :501-509.

[6]FIORUCCI S, RIZZO G, DONINI A, et al.Targeting famesoid Xreceptor for liver and metabolic disorders[J].Trends Mol Med, 2007, 13 (7) :298-309.

[7]RENGA B, MENCARELLI A, D'AMORE C, et al.Discovery that theonellasterol a marine sponge sterol is a highly selective FXR antagonist that protects against liver injury in cholestasis[J].PLo SOne, 2012, 7 (1) :e30443.

[8]LI G, L GUO G.Farnesoid X receptor, the bile acid sensing nuclear receptor, in liver regeneration[J].Acta Pharm Sin B, 2015, 5 (2) :93-98.

[9]INOKUCHI A, HINOSHITA E, IWAMOTO Y, et al.Enhanced expression of the human multidrug resistance protein 3 by bile salt in human enterocytes.A transcriptional control of a plausible bile acid transporter[J].J Biol Chem, 2001, 276 (50) :46822-46829.

[10]KCK K, FERSLEW BC, NETTERBERG I, et al.Risk factors for development of cholestatic drug-induced liver injury:Inhibition of hepatic basolateral bile acid transporters multidrug resistance-associated proteins 3 and 4[J].Drug Metab Dispos, 2014, 42 (4) :665-674.

[11] FIORUCCI S, RIZZO G, DONINI A, et al.Targeting famesoid Xreceptor for liver and metabolic disorders[J].Trends Mol Med, 2007, 13 (7) :298-309.

[12]TRAUNER M, MEIER PJ, BOYER JL.Molecular pathogenesis of cholestasis[J].N Engl J Med, 1998, 339 (17) :1217-1227.

[13]LI XL, SUN FX, WANG XJ, et al.The effect of compound Yindan decoction in rat model with acute intrahepatic cholestasis[J].Chin J Integr Trad West Med Dig, 2014, 22 (9) :497-500. (in Chinese) 李晓玲, 孙凤霞, 王晓静, 等.复方茵丹汤对大鼠急性肝内胆汁淤积的干预作用[J].中国中西结合消化杂志, 2014, 22 (9) :497-500.

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