AIM： To study the association of three common ABCB11 and ABCC2 polymorphisms （ABCB11： 1331T〉C→V444A; ABCC2： 3563T〉A → V1188E and 4544G 〉A → C1515Y） with intrahepatic cholestasis of pregnancy （ICP） and con...AIM： To study the association of three common ABCB11 and ABCC2 polymorphisms （ABCB11： 1331T〉C→V444A; ABCC2： 3563T〉A → V1188E and 4544G 〉A → C1515Y） with intrahepatic cholestasis of pregnancy （ICP） and contraceptive-induced cholestasis （CIC）. METHODS： ABCB11 and ABCC2 genotyping data were available from four CIC patients and from 42 and 33 ICP patients, respectively. Allele-frequencies of the studied polymorphisms were compared with those in healthy pregnant controls and Caucasian individuals. Furthermore, serum bile acid levels were correlated with the presence or absence of the 1331 C allele. RESULTS： The ABCB11 1331T〉C polymorphism was significantly more frequent in cholestatic patients than in pregnant controls： C allele 76.2% （CI, 58.0-94.4） vs 51.3% （CI 35.8-66.7）, respectively （P = 0.0007）; and CC allele 57.1% （CI 36.0-78.3） vs 20% （CI 7.6-32.4）, respectively （P = 0.0065）. All four CIC patients were homozygous carriers of the C allele. In contrast, none of the studied ABCC2 polymorphism was overrepresented in ICP or CIC patients. Higher serum bile acid levels were found in carriers of the 1331CC genotype compared to carriers of the TT genotype. CONCLUSION： Our data support a role for the ABCB11 1331T〉C polymorphism as a susceptibility factor for the development of estrogen-induced cholestasis, whereas no such association was found for ABCC2. Serum bile acid and 7-glutamyl transferase levels might help to distinguish ABCB4- and ABCB11-related forms of ICP and CIC.展开更多
To investigate the relation of two different mutations to the outcome of partial external biliary diversion(PEBD)in severe bile salt export pump(BSEP)deficiency.METHODSMutations in the gene encoding BSEP leading to se...To investigate the relation of two different mutations to the outcome of partial external biliary diversion(PEBD)in severe bile salt export pump(BSEP)deficiency.METHODSMutations in the gene encoding BSEP leading to severe BSEP deficiency in two unrelated patients were identified by genomic sequencing.Native liver biopsies and transiently transfected human embryonic kidney(HEK)293 cells expressing either wild-type or mutated BSEP were subjected to immunofluorescence analysis to assess BSEP transporter localization.Bile acid profiles of patient and control bile samples were generated by ultra-performance liquid chromatography-tandem mass spectrometry.Wild-type and mutant BSEP transport of[<sup>3</sup>H]-labeled taurocholate(TC)and taurochenodeoxycholate(TCDC)was assessed by vesicular transport assays.RESULTSA girl(at 2 mo)presented with pruritus,jaundice and elevated serum bile salts(BS).PEBD stabilized liver function and prevented liver transplantation.She was heterozygous for the BSEP deletion p.T919del and the nonsense mutation p.R1235X.At the age of 17 years relative amounts of conjugated BS in her bile were normal,while total BS were less than 3%as compared to controls.An unrelated boy(age 1.5 years)presenting with severe pruritus and elevated serum BS was heterozygous for the same nonsense and another missense mutation,p.G1032R.PEBD failed to alleviate pruritus,eventually necessitating liver transplantation.BS concentration in bile was about 5%of controls.BS were mainly unconjugated with an unusual low amount of chenodeoxycholate derivatives(<5%).The patients’native liver biopsies showed canalicular BSEP expression.Both BSEP p.T919del and p.G1032R were localized in the plasma membrane in HEK293 cells.In vitro transport assays showed drastic reduction of transport by both mutations.Using purified recombinant BSEP as quantifiable reference,per-molecule transport rates for TC and TCDC were determined to be 3 and 2 BS molecules per wild-type BSEP transporter per minute,respectively.CONCLUSIONIn summary,our findings suggest that residual function of BSEP as well as substrate specificity influence the therapeutic effectiveness of PEBD in progressive familial intrahepatic cholestasis type 2(PFIC-2).展开更多
Progressive familial intrahepatic cholestasis type 2(PFIC2),also known as bile salt export pump(BSEP)deficiency disease,is a rare autosomal recessive inherited liver disease caused by mutations in the ABCB11 gene(loca...Progressive familial intrahepatic cholestasis type 2(PFIC2),also known as bile salt export pump(BSEP)deficiency disease,is a rare autosomal recessive inherited liver disease caused by mutations in the ABCB11 gene(located on chromosome 2q24-31),leading to impaired bile secretion.1 Over 200 distinct mutations in the ABCB11 gene have been identified,including missense,nonsense,insertion,deletion,and splice site mutations.1 Compared with other types of PFIC,patients with BSEP deficiency are at a higher risk of progressing to cirrhosis and liver failure.Current treatment options for PFIC2 remain limited and frustrating.Liver transplantation,the only effective intervention,remains the sole definitive treatment for PFIC2.Nevertheless,its application is severely limited by the prohibitive cost and the scarcity of suitable liver donors.Addressing the pathogenesis of PFIC2 poses a significant clinical challenge.However,a recent study has shown that sirolimus may partially restore the bile excretion ability of ABCB11 mutants in abcb11 knockout Zebrafish models.2 This case report describes a patient with BSEP deficiency disease who responded favorably to sirolimus treatment.展开更多
BACKGROUND Benign recurrent intrahepatic cholestasis(BRIC)is a rare autosomal recessive liver disease,causing episodic cholestasis with intense pruritus.This case report highlights the effectiveness of early plasmaphe...BACKGROUND Benign recurrent intrahepatic cholestasis(BRIC)is a rare autosomal recessive liver disease,causing episodic cholestasis with intense pruritus.This case report highlights the effectiveness of early plasmapheresis as a therapeutic option for BRIC type 2,offering rapid symptom relief and early termination of cholestatic episodes.It contributes to the limited evidence supporting plasmapheresis as a treatment for BRIC flares resistant to conventional therapies.CASE SUMMARY A 43-year-old male with BRIC type 2 presented with fatigue,jaundice,and severe pruritus,triggered by a recent mild severe acute respiratory syndrome coronavirus 2 infection.Laboratory results confirmed cholestasis with elevated bilirubin and alkaline phosphatase.First-line pharmacological treatments,including cholestyramine and rifampicin,failed.Endoscopic nasobiliary drainage was ineffective,prompting initiation of plasmapheresis.This intervention rapidly relieved pruritus,with complete biochemical normalisation after 11 sessions.Two years later,a similar episode occurred,and early reinitiation of plasmapheresis led to symptom resolution within two sessions and biochemical recovery within two weeks.The patient tolerated the procedure well,with no adverse effects observed.Follow-up showed no signs of cholestasis recurrence.CONCLUSION Plasmapheresis is a safe and effective option for therapy-refractory BRIC type 2,particularly when initiated early in cholestasis.展开更多
目的利福平(Rifampicin,RIF)具有肝毒性,但其机制尚不清楚。本研究在RIF诱导的肝内胆汁淤积小鼠中,探讨RIF对肝细胞胆汁酸转运体胆汁酸输出泵(bile salt exportpump,Bsep)和多药抵抗相关蛋白-2(multidrug resistance-associated protein...目的利福平(Rifampicin,RIF)具有肝毒性,但其机制尚不清楚。本研究在RIF诱导的肝内胆汁淤积小鼠中,探讨RIF对肝细胞胆汁酸转运体胆汁酸输出泵(bile salt exportpump,Bsep)和多药抵抗相关蛋白-2(multidrug resistance-associated protein-2,Mrp2)表达和定位影响。方法 48只♀ICR小鼠随机分为4组,RIF1wk组:经灌胃给予RIF(200mg.kg-1.d-1),连续1周,于末次给药后6h取材;RIF6h组:单次灌胃给予RIF(200mg.kg-1)后6h取材;RIF1周对照组(CON1wk)与RIF6h对照组(CON6h):经灌胃给予等容积生理盐水。所有小鼠均收集血液和肝组织,常规生化检测血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、总胆红素(TB)和结合胆红素(DB),并检测小鼠血清和肝组织总胆汁酸(TBA)水平。HE染色分析肝组织病理改变。RT-PCR测定肝脏肝细胞胆汁酸转运体Bsep和Mrp2mRNA表达。免疫荧光法分析Bsep和Mrp2在肝细胞的位置。结果给予RIF1周后,小鼠血清TB由(1.25±0.69)μmol.L-1上升至(65.73±12.08)μmol.L-1,上升近70倍,DB由(0.77±0.40)μmol.L-1上升至(53.33±12.43)μmol.L-1,上升约80倍,ALP由(110.2±13.8)U.L-1上升至(279.5±80.4)U.L-1,上升约1.5倍,TBA由(3.15±0.89)μmol.L-1上升至(13.54±6.51)μmol.L-1,上升约5倍并伴有血清ALT和AST轻度升高;肝脏组织TBA由(0.15±0.04)μmol.g-1liver上升至(0.30±0.19)μmol.g-1liver,上升约2倍;肝脏组织HE染色显示肝细胞出现脂肪变性、轻度坏死和炎症。单次给予RIF6h后血清TB、DB、ALP、ALT、AST和TBA明显上升,但未观察到小鼠肝脏组织病理发生改变。免疫荧光分析显示,给予小鼠RIF1wk与单次给予RIF6h后肝细胞中Bsep和Mrp2的定位发生了改变。而无论单次给予RIF还是连续给药1周,肝细胞Bsep和Mrp2mRNA表达水平均未发生变化。结论肝细胞胆汁酸转运体Bsep和Mrp2定位改变可能是RIF诱发肝内胆汁淤积的重要机制。展开更多
目的通过考察茵栀黄颗粒(茵陈、栀子、金银花和黄芩)对肝细胞膜转运体多药耐药相关蛋白2(Multidrugresistance-associated protein 2,Mrp2/ABCC2)、Na+-牛黄胆酸共转运多肽(Na+/taurocholate cotransporter,Ntcp/SLC10A1)及胆盐输出泵(B...目的通过考察茵栀黄颗粒(茵陈、栀子、金银花和黄芩)对肝细胞膜转运体多药耐药相关蛋白2(Multidrugresistance-associated protein 2,Mrp2/ABCC2)、Na+-牛黄胆酸共转运多肽(Na+/taurocholate cotransporter,Ntcp/SLC10A1)及胆盐输出泵(Bile salt export pump,Bsep/ABCB11)表达的影响。方法胆管结扎术制备胆汁淤积大鼠模型;生化检测和HE染色,观察茵栀黄颗粒的干预效果;流式细胞仪测定肝脏转运体Ntcp、Bsep和Mrp2的表达。结果与模型组相比,茵栀黄组肝细胞的脂肪病变和水肿较模型组显著减轻(P<0.05),Mrp2的表达显著升高(P<0.01);但Ntcp、Bsep的表达无明显变化。结论肝细胞膜转运体Mrp2的表达升高可能是茵栀黄颗粒的退黄利胆分子机制之一。展开更多
基金Supported by Grants from the Gebert Rüf Foundation, the Forschungskredit of the University Zurichthe Swiss National Science Foundation, Grants PP00B-108511/1 and 31-64140.00
文摘AIM： To study the association of three common ABCB11 and ABCC2 polymorphisms （ABCB11： 1331T〉C→V444A; ABCC2： 3563T〉A → V1188E and 4544G 〉A → C1515Y） with intrahepatic cholestasis of pregnancy （ICP） and contraceptive-induced cholestasis （CIC）. METHODS： ABCB11 and ABCC2 genotyping data were available from four CIC patients and from 42 and 33 ICP patients, respectively. Allele-frequencies of the studied polymorphisms were compared with those in healthy pregnant controls and Caucasian individuals. Furthermore, serum bile acid levels were correlated with the presence or absence of the 1331 C allele. RESULTS： The ABCB11 1331T〉C polymorphism was significantly more frequent in cholestatic patients than in pregnant controls： C allele 76.2% （CI, 58.0-94.4） vs 51.3% （CI 35.8-66.7）, respectively （P = 0.0007）; and CC allele 57.1% （CI 36.0-78.3） vs 20% （CI 7.6-32.4）, respectively （P = 0.0065）. All four CIC patients were homozygous carriers of the C allele. In contrast, none of the studied ABCC2 polymorphism was overrepresented in ICP or CIC patients. Higher serum bile acid levels were found in carriers of the 1331CC genotype compared to carriers of the TT genotype. CONCLUSION： Our data support a role for the ABCB11 1331T〉C polymorphism as a susceptibility factor for the development of estrogen-induced cholestasis, whereas no such association was found for ABCC2. Serum bile acid and 7-glutamyl transferase levels might help to distinguish ABCB4- and ABCB11-related forms of ICP and CIC.
基金Supported by the German-Research Foun-dation-through the Clin-ical Research Group KFO217“Hepatobiliary tran-sport an-d liver diseases”the Collaborative Research Cen-tre 974“Commun-ication-an-d Systemic Relevan-ce in-Liver Damage an-d Regen-eration-”
文摘To investigate the relation of two different mutations to the outcome of partial external biliary diversion(PEBD)in severe bile salt export pump(BSEP)deficiency.METHODSMutations in the gene encoding BSEP leading to severe BSEP deficiency in two unrelated patients were identified by genomic sequencing.Native liver biopsies and transiently transfected human embryonic kidney(HEK)293 cells expressing either wild-type or mutated BSEP were subjected to immunofluorescence analysis to assess BSEP transporter localization.Bile acid profiles of patient and control bile samples were generated by ultra-performance liquid chromatography-tandem mass spectrometry.Wild-type and mutant BSEP transport of[<sup>3</sup>H]-labeled taurocholate(TC)and taurochenodeoxycholate(TCDC)was assessed by vesicular transport assays.RESULTSA girl(at 2 mo)presented with pruritus,jaundice and elevated serum bile salts(BS).PEBD stabilized liver function and prevented liver transplantation.She was heterozygous for the BSEP deletion p.T919del and the nonsense mutation p.R1235X.At the age of 17 years relative amounts of conjugated BS in her bile were normal,while total BS were less than 3%as compared to controls.An unrelated boy(age 1.5 years)presenting with severe pruritus and elevated serum BS was heterozygous for the same nonsense and another missense mutation,p.G1032R.PEBD failed to alleviate pruritus,eventually necessitating liver transplantation.BS concentration in bile was about 5%of controls.BS were mainly unconjugated with an unusual low amount of chenodeoxycholate derivatives(<5%).The patients’native liver biopsies showed canalicular BSEP expression.Both BSEP p.T919del and p.G1032R were localized in the plasma membrane in HEK293 cells.In vitro transport assays showed drastic reduction of transport by both mutations.Using purified recombinant BSEP as quantifiable reference,per-molecule transport rates for TC and TCDC were determined to be 3 and 2 BS molecules per wild-type BSEP transporter per minute,respectively.CONCLUSIONIn summary,our findings suggest that residual function of BSEP as well as substrate specificity influence the therapeutic effectiveness of PEBD in progressive familial intrahepatic cholestasis type 2(PFIC-2).
基金supported by the 5010 Cultivation Program of Clinical Research of Sun Yat-Sen University(No.2018024)Guangzhou Science and Technology Plan Project(2023A03J0807).
文摘Progressive familial intrahepatic cholestasis type 2(PFIC2),also known as bile salt export pump(BSEP)deficiency disease,is a rare autosomal recessive inherited liver disease caused by mutations in the ABCB11 gene(located on chromosome 2q24-31),leading to impaired bile secretion.1 Over 200 distinct mutations in the ABCB11 gene have been identified,including missense,nonsense,insertion,deletion,and splice site mutations.1 Compared with other types of PFIC,patients with BSEP deficiency are at a higher risk of progressing to cirrhosis and liver failure.Current treatment options for PFIC2 remain limited and frustrating.Liver transplantation,the only effective intervention,remains the sole definitive treatment for PFIC2.Nevertheless,its application is severely limited by the prohibitive cost and the scarcity of suitable liver donors.Addressing the pathogenesis of PFIC2 poses a significant clinical challenge.However,a recent study has shown that sirolimus may partially restore the bile excretion ability of ABCB11 mutants in abcb11 knockout Zebrafish models.2 This case report describes a patient with BSEP deficiency disease who responded favorably to sirolimus treatment.
文摘BACKGROUND Benign recurrent intrahepatic cholestasis(BRIC)is a rare autosomal recessive liver disease,causing episodic cholestasis with intense pruritus.This case report highlights the effectiveness of early plasmapheresis as a therapeutic option for BRIC type 2,offering rapid symptom relief and early termination of cholestatic episodes.It contributes to the limited evidence supporting plasmapheresis as a treatment for BRIC flares resistant to conventional therapies.CASE SUMMARY A 43-year-old male with BRIC type 2 presented with fatigue,jaundice,and severe pruritus,triggered by a recent mild severe acute respiratory syndrome coronavirus 2 infection.Laboratory results confirmed cholestasis with elevated bilirubin and alkaline phosphatase.First-line pharmacological treatments,including cholestyramine and rifampicin,failed.Endoscopic nasobiliary drainage was ineffective,prompting initiation of plasmapheresis.This intervention rapidly relieved pruritus,with complete biochemical normalisation after 11 sessions.Two years later,a similar episode occurred,and early reinitiation of plasmapheresis led to symptom resolution within two sessions and biochemical recovery within two weeks.The patient tolerated the procedure well,with no adverse effects observed.Follow-up showed no signs of cholestasis recurrence.CONCLUSION Plasmapheresis is a safe and effective option for therapy-refractory BRIC type 2,particularly when initiated early in cholestasis.
文摘目的通过考察茵栀黄颗粒(茵陈、栀子、金银花和黄芩)对肝细胞膜转运体多药耐药相关蛋白2(Multidrugresistance-associated protein 2,Mrp2/ABCC2)、Na+-牛黄胆酸共转运多肽(Na+/taurocholate cotransporter,Ntcp/SLC10A1)及胆盐输出泵(Bile salt export pump,Bsep/ABCB11)表达的影响。方法胆管结扎术制备胆汁淤积大鼠模型;生化检测和HE染色,观察茵栀黄颗粒的干预效果;流式细胞仪测定肝脏转运体Ntcp、Bsep和Mrp2的表达。结果与模型组相比,茵栀黄组肝细胞的脂肪病变和水肿较模型组显著减轻(P<0.05),Mrp2的表达显著升高(P<0.01);但Ntcp、Bsep的表达无明显变化。结论肝细胞膜转运体Mrp2的表达升高可能是茵栀黄颗粒的退黄利胆分子机制之一。
