肾小管性酸中毒血清抗V-H~+-ATP酶B1/B2亚基抗体的检测
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摘要
研究目的
免疫因素在肾小管性酸中毒发病中的作用日渐受到关注。已有研究报道肾小管性酸中毒患者血清中存在抗肾小管上皮细胞自身抗体,但这些抗体的具体种类尚未明确。
本实验旨在检测肾小管性酸中毒患者血清中是否存在抗v型氢离子腺苷三磷酸酶(v-H~+-ATP酶)B1亚基及B2亚基自身抗体。
研究方法
研究对象:肾小管性酸中毒患者11例,其中10例合并干燥综合征:单纯干燥综合征患者8例;健康对照8例。所有患者均为未治疗的新发或复发病例。取各研究对象全血利用流式细胞技术进行免疫学检查,部分血清进行生化等指标检测,部分血清用于间接免疫荧光检测自身抗体。
先确定患者血清中是否存在抗肾小管上皮细胞抗体,以及观察v-H~+-ATP酶B1亚基和B2亚基在正常人肾脏组织的表达特点。在正常人肾脏组织石蜡切片上,加肾小管性酸中毒患者血清原液、单纯干燥综合征患者血清原液、健康对照血清原液、抗v-H~+-ATP酶B1亚基抗体或抗v-H~+-ATP酶B2亚基抗体作为一抗孵育4℃过夜,以异硫氰酸荧光素(Fluoresceinisothiocynate,FITC,绿色荧光)-山羊抗人免疫球蛋白(Immunoglobulin,IgG)孵育,标记血清中的抗肾小管上皮细胞自身抗体,或四甲基异硫氰酸罗丹明(tetramethylrhodamineisothiocyanate,TRITC,红色荧光)-兔抗山羊IgG孵育,标记v-H~+-ATP酶B1亚基或B2亚基,4,6-联脒-2-苯基吲哚(4,6-diamidino-2-phenyl-indolediacetare,DAPI,蓝色荧光)标记细胞核并封片。结果阳性者,进一步稀释相应的一抗,重复上述实验,直至结果为阴性。并为以下间接免疫荧光双标记实验选取适当的稀释比例。
为检测肾小管性酸中毒患者血清中是否存在抗v-H~+-ATP酶B1亚基及B2亚基自身抗体,先用肾小管性酸中毒患者血清原液、单纯干燥综合征患者血清原液或健康对照血清原液与正常人肾脏组织石蜡切片孵育4℃过夜,然后加不同稀释度的山羊抗人v-H~+-ATP酶B1亚基抗体孵育4℃过夜,再先后加TRITC-兔抗山羊IgG及FITC-山羊抗人IgG荧光二抗避光孵育,DAPI标记细胞核并封片。结果阳性者,将血清按不同比例稀释,重复上述实验,直至结果阴性。血清中抗v-H~+-ATP酶B2亚基自身抗体的检测同抗v-H~+-ATP酶B1亚基自身抗体的检测。
每次实验均设阴性对照。
结果
一般临床指标检查结果:与单纯干燥综合征患者和健康对照相比较,肾小管性酸中毒患者的血清氯水平明显升高,血清二氧化碳明显下降,尿pH明显升高,流式细胞学检查、免疫球蛋白及补体检查结果均提示免疫学变化。
血清抗肾小管上皮细胞自身抗体检测结果:11例肾小管性酸中毒患者中,有6例患者肾小管上皮细胞内呈明亮的绿色荧光,提示抗肾小管上皮细胞抗体阳性;当把自身抗体阳性的血清逐渐稀释时,其绿色荧光的强度呈不同程度地逐渐减弱,提示随着血清的稀释,自身抗体的浓度逐渐下降;而8例单纯干燥综合征患者及8例健康对照的肾小管上皮细胞内无明显绿色荧光,提示其血清中抗肾小管上皮细胞自身抗体阴性。
v-H~+-ATP酶B1亚基及B2亚基在正常肾脏组织的表达结果:标记v-H~+-ATP酶B1亚基的红色荧光在肾脏中呈闰细胞特异性散在分布,标记v-H~+-ATP酶B2亚基的红色荧光在肾小管上皮细胞内沿管腔面呈连续的线状分布。
抗v-H~+-ATP酶B1亚基及B2亚基自身抗体检测结果:11例肾小管性酸中毒患者中,有6例患者标记v-H~+-ATP酶B1亚基的红色荧光强度明显减弱,提示其血清对于抗v-H~+-ATP酶B1亚基抗体有明显的阻断作用;当把自身抗体阳性患者的血清逐渐稀释,标记v-H~+-ATP酶B1亚基的红色荧光强度逐渐增强,提示随着血清的稀释,其对于抗v-H~+-ATP酶B1亚基抗体的阻断作用呈不同程度的减弱,提示不同稀释度血清的抗体效价强度不同;而8例单纯干燥综合征患者和8例健康对照血清则无上述阻断作用。抗v-H~+-ATP酶B2亚基自身抗体的检测结果同抗v-H~+-ATP酶B1亚基自身抗体检测结果。以上结果提示肾小管性酸中毒血清中存在抗v-H~+-ATP酶B1亚基及B2亚基自身抗体。
结论
肾小管酸中毒患者血清中存在抗肾小管上皮细胞自身抗体,阳性率为6/11;肾小管性酸中毒患者血清中存在抗v-H~+-ATP酶B1亚基及B2亚基自身抗体,阳性率均为6/11:肾小管性酸中毒患者血清中的抗v-H~+-ATP酶B1亚基及B2亚基自身抗体是抗肾小管上皮细胞自身抗体中的两种。
Objective
Autoimmunity plays an important role in renal tubular acidosis(RTA).The presence of autoantibodies against renal tubular epithelial cells has been found in patients with RTA,but the detailed description of these autoantibodies is current lacking.
Our aims of this study was to detect a presence or absence of autoantibodies against B1 and/or B2 subunits of vacuolar H~+-adenosine triphosphatease(v-H~+-ATPase) in sera of patients diagnosed with RTA.
Methods
Eleven untreated patients diagnosed with RTA(10 of these cases were associated with Sjogren syndrome),eight untreated patients with simple Sjogren syndrome(sSS)and eight healthy controls(HC)were enrolled in the study.Part of the fasting whole blood was taken out for measuring immuno-function by flow cytometry, and other serum was for detecting serum biochemistry,and autoantibodies presence or absence by indirect immunofluorescence assay.
Firstly,to detect a presence or absence of serum autoantibodies against renal tubular epithelial cells and observe the expression of B 1 subunit and B2 subunits of v-H~+-ATPase in normal renal tissue of human,paraffin sections of normal human renal tissue were incubated with sera of RTA,sSS,HC or anti- B1 or B2 subunits of v-H~+-ATPase antibodies overnight at 4℃,then autoantibodies against renal tubular epithelial cells in sera were conjugated with fluorescein isothiocyanate(FITC,green color)-labeled goat-anti-human immunoglobulin G(IgG),while B1 or B2 subunits of V-H~+-ATPase were conjugated with tetramethylrhodamine isothiocyanate(TRITC, red color)-labeled rabbit-anti-goat IgG.Nuclei were stained with 4,6-diamino-2-phenyl indole(DAPI,blue color).For each positive sample,the sera or anti-B1 or B2 subunits of V-H~+-ATPase antibodies were diluted to repeat the trial until negative results were obtained.
To confirm a presence or absence of autoantibodies against B1 and B2 subunits of v-H~+-ATPase in sera of RTA,paraffin sections of normal human renal tissue were firstly incubated with sera of RTA,sSS or HC overnight at 4℃,and with goat-anti-v-H~+-ATPase B1 subunit antibodies followed later overnight at 4℃.Then,the sections were incubated with TRITC-labeled rabbit-anti-goat secondary antibodies for B subunit of v-H~+- ATPase assay,and with FITC-labeled goat-anti-human secondary antibodies for autoantibodies against renal tubular epithelial cells.Finally,nuclei were stained with DAPI.For each positive sample,we diluted the sera and repeated the trial until negative.Similarly,detection of autoantibodies against B2 subunit of v-H~+-ATPase was performed by the same protocol as that against B1 subunit of v-H~+-ATPase assay described previously.
Results
Results of general indicators:Compared with patients with sSS and HC,results in sera of patients with RTA showed that serum chloride increased,CO2 decreased and urine pH increased.And results of flow cytometry,IG and complements implied immune disorders.
Results of autoantibodies against renal tubular epithelial cells:The sera of 6 cases of all RTA patients showed green fluorescence staining in renal tubular epithelial cells,additional experiments showed that the staining intensity was also reduced following the sera of patients with RTA were diluted.Moreover,none of those sera from patients with either sSS or HC showed green fluorescence in renal tubular epithelial cells,which indicated no autoantibodies against renal tubular epithelial cells in sera of patients with sSS and HC.The results indicated the presence of autoantibodies against renal tubular epithelial cells in sera of patients with RTA.
Results of distribution characteristics of B1 and B2 subunits of v-H~+-ATPase in normal human renal tissue:Red fluorescence representing B1 subunit of v-H~+-ATPase distributed specifically in intercalated cells,while B2 subunit distributed continuously in the cavosurface of renal tubular epithelial cells.
Assay results of autoantibodies against B1 and B2 subunits of v-H~+-ATPase: when sera were incubated prior to anti-B1 subunit antibodies,the red fluorescence intensity showed a decrease in 6 cases of the 11 patients with RTA,which implied that blocking effect of the sera of patients with RTA to anti-B1 subunit of v-H~+-ATPase antibodies was occurred.The sera-induced blocking effects were reduced following the sera of patients with RTA diluted.While none of sera from these patients with either sSS or HC showed the blocking effect like that.The assay results of autoantibodies against B2 subunit of v-H~+-ATPase were as the same as that of B1 subunit.
Conclusion
There are autoantibodies against renal tubular epithelial cells in sera of patients with RTA,and the positive ratio is 6/11.There are autoantibodies against B1 and B2 subunits of v-H~+-ATPase in sera of patients with RTA,and the positive ratio are both 6/11.Autoantibodies against B1 and B2 subunits of v-H~+-ATPase are two kinds of autoantibodies against renal tubular epithelial cells in sera of patients with RTA.
免疫因素在肾小管性酸中毒发病中的作用日渐受到关注。已有研究报道肾小管性酸中毒患者血清中存在抗肾小管上皮细胞自身抗体,但这些抗体的具体种类尚未明确。
本实验旨在检测肾小管性酸中毒患者血清中是否存在抗v型氢离子腺苷三磷酸酶(v-H~+-ATP酶)B1亚基及B2亚基自身抗体。
研究方法
研究对象:肾小管性酸中毒患者11例,其中10例合并干燥综合征:单纯干燥综合征患者8例;健康对照8例。所有患者均为未治疗的新发或复发病例。取各研究对象全血利用流式细胞技术进行免疫学检查,部分血清进行生化等指标检测,部分血清用于间接免疫荧光检测自身抗体。
先确定患者血清中是否存在抗肾小管上皮细胞抗体,以及观察v-H~+-ATP酶B1亚基和B2亚基在正常人肾脏组织的表达特点。在正常人肾脏组织石蜡切片上,加肾小管性酸中毒患者血清原液、单纯干燥综合征患者血清原液、健康对照血清原液、抗v-H~+-ATP酶B1亚基抗体或抗v-H~+-ATP酶B2亚基抗体作为一抗孵育4℃过夜,以异硫氰酸荧光素(Fluoresceinisothiocynate,FITC,绿色荧光)-山羊抗人免疫球蛋白(Immunoglobulin,IgG)孵育,标记血清中的抗肾小管上皮细胞自身抗体,或四甲基异硫氰酸罗丹明(tetramethylrhodamineisothiocyanate,TRITC,红色荧光)-兔抗山羊IgG孵育,标记v-H~+-ATP酶B1亚基或B2亚基,4,6-联脒-2-苯基吲哚(4,6-diamidino-2-phenyl-indolediacetare,DAPI,蓝色荧光)标记细胞核并封片。结果阳性者,进一步稀释相应的一抗,重复上述实验,直至结果为阴性。并为以下间接免疫荧光双标记实验选取适当的稀释比例。
为检测肾小管性酸中毒患者血清中是否存在抗v-H~+-ATP酶B1亚基及B2亚基自身抗体,先用肾小管性酸中毒患者血清原液、单纯干燥综合征患者血清原液或健康对照血清原液与正常人肾脏组织石蜡切片孵育4℃过夜,然后加不同稀释度的山羊抗人v-H~+-ATP酶B1亚基抗体孵育4℃过夜,再先后加TRITC-兔抗山羊IgG及FITC-山羊抗人IgG荧光二抗避光孵育,DAPI标记细胞核并封片。结果阳性者,将血清按不同比例稀释,重复上述实验,直至结果阴性。血清中抗v-H~+-ATP酶B2亚基自身抗体的检测同抗v-H~+-ATP酶B1亚基自身抗体的检测。
每次实验均设阴性对照。
结果
一般临床指标检查结果:与单纯干燥综合征患者和健康对照相比较,肾小管性酸中毒患者的血清氯水平明显升高,血清二氧化碳明显下降,尿pH明显升高,流式细胞学检查、免疫球蛋白及补体检查结果均提示免疫学变化。
血清抗肾小管上皮细胞自身抗体检测结果:11例肾小管性酸中毒患者中,有6例患者肾小管上皮细胞内呈明亮的绿色荧光,提示抗肾小管上皮细胞抗体阳性;当把自身抗体阳性的血清逐渐稀释时,其绿色荧光的强度呈不同程度地逐渐减弱,提示随着血清的稀释,自身抗体的浓度逐渐下降;而8例单纯干燥综合征患者及8例健康对照的肾小管上皮细胞内无明显绿色荧光,提示其血清中抗肾小管上皮细胞自身抗体阴性。
v-H~+-ATP酶B1亚基及B2亚基在正常肾脏组织的表达结果:标记v-H~+-ATP酶B1亚基的红色荧光在肾脏中呈闰细胞特异性散在分布,标记v-H~+-ATP酶B2亚基的红色荧光在肾小管上皮细胞内沿管腔面呈连续的线状分布。
抗v-H~+-ATP酶B1亚基及B2亚基自身抗体检测结果:11例肾小管性酸中毒患者中,有6例患者标记v-H~+-ATP酶B1亚基的红色荧光强度明显减弱,提示其血清对于抗v-H~+-ATP酶B1亚基抗体有明显的阻断作用;当把自身抗体阳性患者的血清逐渐稀释,标记v-H~+-ATP酶B1亚基的红色荧光强度逐渐增强,提示随着血清的稀释,其对于抗v-H~+-ATP酶B1亚基抗体的阻断作用呈不同程度的减弱,提示不同稀释度血清的抗体效价强度不同;而8例单纯干燥综合征患者和8例健康对照血清则无上述阻断作用。抗v-H~+-ATP酶B2亚基自身抗体的检测结果同抗v-H~+-ATP酶B1亚基自身抗体检测结果。以上结果提示肾小管性酸中毒血清中存在抗v-H~+-ATP酶B1亚基及B2亚基自身抗体。
结论
肾小管酸中毒患者血清中存在抗肾小管上皮细胞自身抗体,阳性率为6/11;肾小管性酸中毒患者血清中存在抗v-H~+-ATP酶B1亚基及B2亚基自身抗体,阳性率均为6/11:肾小管性酸中毒患者血清中的抗v-H~+-ATP酶B1亚基及B2亚基自身抗体是抗肾小管上皮细胞自身抗体中的两种。
Objective
Autoimmunity plays an important role in renal tubular acidosis(RTA).The presence of autoantibodies against renal tubular epithelial cells has been found in patients with RTA,but the detailed description of these autoantibodies is current lacking.
Our aims of this study was to detect a presence or absence of autoantibodies against B1 and/or B2 subunits of vacuolar H~+-adenosine triphosphatease(v-H~+-ATPase) in sera of patients diagnosed with RTA.
Methods
Eleven untreated patients diagnosed with RTA(10 of these cases were associated with Sjogren syndrome),eight untreated patients with simple Sjogren syndrome(sSS)and eight healthy controls(HC)were enrolled in the study.Part of the fasting whole blood was taken out for measuring immuno-function by flow cytometry, and other serum was for detecting serum biochemistry,and autoantibodies presence or absence by indirect immunofluorescence assay.
Firstly,to detect a presence or absence of serum autoantibodies against renal tubular epithelial cells and observe the expression of B 1 subunit and B2 subunits of v-H~+-ATPase in normal renal tissue of human,paraffin sections of normal human renal tissue were incubated with sera of RTA,sSS,HC or anti- B1 or B2 subunits of v-H~+-ATPase antibodies overnight at 4℃,then autoantibodies against renal tubular epithelial cells in sera were conjugated with fluorescein isothiocyanate(FITC,green color)-labeled goat-anti-human immunoglobulin G(IgG),while B1 or B2 subunits of V-H~+-ATPase were conjugated with tetramethylrhodamine isothiocyanate(TRITC, red color)-labeled rabbit-anti-goat IgG.Nuclei were stained with 4,6-diamino-2-phenyl indole(DAPI,blue color).For each positive sample,the sera or anti-B1 or B2 subunits of V-H~+-ATPase antibodies were diluted to repeat the trial until negative results were obtained.
To confirm a presence or absence of autoantibodies against B1 and B2 subunits of v-H~+-ATPase in sera of RTA,paraffin sections of normal human renal tissue were firstly incubated with sera of RTA,sSS or HC overnight at 4℃,and with goat-anti-v-H~+-ATPase B1 subunit antibodies followed later overnight at 4℃.Then,the sections were incubated with TRITC-labeled rabbit-anti-goat secondary antibodies for B subunit of v-H~+- ATPase assay,and with FITC-labeled goat-anti-human secondary antibodies for autoantibodies against renal tubular epithelial cells.Finally,nuclei were stained with DAPI.For each positive sample,we diluted the sera and repeated the trial until negative.Similarly,detection of autoantibodies against B2 subunit of v-H~+-ATPase was performed by the same protocol as that against B1 subunit of v-H~+-ATPase assay described previously.
Results
Results of general indicators:Compared with patients with sSS and HC,results in sera of patients with RTA showed that serum chloride increased,CO2 decreased and urine pH increased.And results of flow cytometry,IG and complements implied immune disorders.
Results of autoantibodies against renal tubular epithelial cells:The sera of 6 cases of all RTA patients showed green fluorescence staining in renal tubular epithelial cells,additional experiments showed that the staining intensity was also reduced following the sera of patients with RTA were diluted.Moreover,none of those sera from patients with either sSS or HC showed green fluorescence in renal tubular epithelial cells,which indicated no autoantibodies against renal tubular epithelial cells in sera of patients with sSS and HC.The results indicated the presence of autoantibodies against renal tubular epithelial cells in sera of patients with RTA.
Results of distribution characteristics of B1 and B2 subunits of v-H~+-ATPase in normal human renal tissue:Red fluorescence representing B1 subunit of v-H~+-ATPase distributed specifically in intercalated cells,while B2 subunit distributed continuously in the cavosurface of renal tubular epithelial cells.
Assay results of autoantibodies against B1 and B2 subunits of v-H~+-ATPase: when sera were incubated prior to anti-B1 subunit antibodies,the red fluorescence intensity showed a decrease in 6 cases of the 11 patients with RTA,which implied that blocking effect of the sera of patients with RTA to anti-B1 subunit of v-H~+-ATPase antibodies was occurred.The sera-induced blocking effects were reduced following the sera of patients with RTA diluted.While none of sera from these patients with either sSS or HC showed the blocking effect like that.The assay results of autoantibodies against B2 subunit of v-H~+-ATPase were as the same as that of B1 subunit.
Conclusion
There are autoantibodies against renal tubular epithelial cells in sera of patients with RTA,and the positive ratio is 6/11.There are autoantibodies against B1 and B2 subunits of v-H~+-ATPase in sera of patients with RTA,and the positive ratio are both 6/11.Autoantibodies against B1 and B2 subunits of v-H~+-ATPase are two kinds of autoantibodies against renal tubular epithelial cells in sera of patients with RTA.
引文
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12.PAUL A.STEHBERGER,NICOLE SCHULZ,KARIN E.FINBERG,et al.Localization and Regulation of the ATP6VOA4(a4)Vacuolar H~+-ATPase Subunit Defective in an Inherited Form of Distal Renal Tubular Acidosis[J].J Am Soc Nephrol,2003,14:3027-3038
13.郑法雷.肾小管酸中毒的诊断与治疗[J].医师进修杂志,2003,26(5):1-3
14.Eriksson P,Denneberg T,Enestrom S,et al.Urolithoasis and distal renal tubular acidosis preceding primary Sjogren syndrome:a retrospective study 5-53 years after the presentation of urolithiasis[J].J Intern Med,1996,239:438-488
15.Lai LW,Chan DM,Erichson RP,et al.Correction of renal tubular acidosis in carbonic anyhdras Ⅱ-deficiant mice with gene therapy[J].J Clin Invest,1998,101:1305-1325
16.杨继红.肾小管酸中毒的临床表现与治疗[J].新医学,2007,38(9):570-572
17.张乃峥主编.临床风湿病学[M].上海:上海科学技术出版社,1999,287-298
18.Aasarqd K,Haga HJ,Berg KJ,et al.Renal involvement in primary Sjogren's syndrome[J].Q J Med,2000,93:297-304
19.董怡,张乃峥.干燥综合征的肾脏损害[J].中华内科杂志,1988,27:162-164
20.董德长主编.实用肾脏病学[M].上海:上海科学技术出版社,1999,1036-1045
21.Pertovaara M,Korpela M,Kouri T,et al.The occurrence of renal involvement in primary Sjogren's syndrome:a study of 78 patients[J].Rheumatology,1999,38:1113- 1120
22.Howard E.Corey·Alfredo Vallo'Juan Rodr_guez-Soriano.An analysis of renal tubular acidosis by the Stewart method[J].Pediatr Nephrol,2006,21:206-211
23.廖二元,莫朝晖主编.内分泌学..第2版[M].北京:人民卫生出版社,2007,601-604
24.杨华彬.Graves病并远端肾小管性酸中毒与周期性瘫痪[J].临床误诊误治,2004,17(12):905-906
25.钱桐荪.远端肾小管性酸中毒的病因和临床表现[J].新医学,2007,38(9):568-570
26.Fiona E.Karet,Karin E.Finberg,Raoul D.Nelson,et al.Mutations in the gene encoding B1 subunit of H~+-ATPase cause renal tubular acidosis with sensorineural deafness[J].Nature Genetics,1999,21(1):84-9027.Ya Su,Aiwu Zhou,Rafia S.Al-Lamki,et al.The a-subunit of the v-type H~+-AYPase interact with phosphofructokinase-1 in humans[J].The journal of biological chemistry,2003,278(22):20013-20018
28.Cremers C.W.,Di Pietro A.,et al.Mutations in the gene encoding B1 subunit of H~+-ATPase cause RTA with sensorineural deafness[J].Nature genetics,1999,21:84-90
29.Carsten A.Wagner,Karin E.Finberg,Sylvie Breton,et al.Renal vacuolar H~+-ATPase[J].Physiol rev.2004,84(4):1263-314
30.Karin E.Finberga,Carsten A.Wagnerb,Paul A.Stehbergerb,et al.Molecular cloning and characterization of Atp6v1b1,the murine vacuolar H~+-ATPase B1-subunit[J].Gene,2003,318:25-34
31.Karin E.Finberg,Carsten A.Wagner,Matthew A.Bailey,et al.The B1-subunit of the H~+- AYPase is required for maximal urinary acidification[J].PNAS,2005,102(38)13616-13621
32.Florina Rothenberger,Ana Velic,Paul A.Stehberger,et al.Angiotensin Ⅱstimulates vacuolar H+ -ATPase activity in renal acid-secretory intercalated cells from the outer medullary collecting duct[J].J Am Soc Nephrol,2007,18:2085-2093
33.G Li,Q Yang,S Krishnan,et al.A novel cellular survival factor - the B2 subunit of vacuolar H~+-ATPase inhibits apoptosis[J].Cell death and differentiation,2006,13:2109-2117
34.陈慰峰主编.医学免疫学..第三版[M].北京:人民卫生出版社,2002,209-210
35.Fumi Takemoto,Junichi Hoshino,Naoki Sawa,et al.Autoantibodies against carbonic anhydrase Ⅱ are increased in renal tubular acidosis associated with Sjogren syndrome[J].The American Journal of Medicine,2005,118:181-184
36.Kohriyama K,Katayama Y,Tsurusako Y.Relationship between primary Sjogren's syndrome and autoimmune thyroid disease[L].Nippon Rinsho,1999,57:1878-1881
37.任红,陈楠,陈晓农,等.干燥综合征合并肾脏损害147例临床病理及随访情况[J].中华风湿病学杂志,2005,9(6):351-353
38.周叶青,胡曼东.舍格伦综合症-合并肾小管酸中毒的免疫学研究[J].口腔医学,2001,21(4):197-198
39.江永娣,陈楠,王朝晖,等.肾小管性酸中毒115例临床分析[J].中华内分泌代谢杂志,1998,14:155-158
40.K.W JOO,U,S JEON J S HAN,S KIM.JS.LEE,et al.Absence of H~+-ATPase in the intercalated cells of renal tissues in classic distal renal tubular acidosis[J].Clinical nephrology,1998,149(4):226-231
41.JIN SUK HAN,GHEUN-HO KIM,JIN KIM,et al.Secretory-Defect Distal Renal Tubular Acidosis Is Associatedwith Transporter Defect in H~+-AYPase and AnionExchanger-1[J].J Am Soc Nephrol,2002,13:1425-1432
2. JUAN RODRI'GUEZ SORIANO. Renal Tubular Acidosis: The Clinical Entity[J]. J Am Soc Nephrol,2002,13: 2160-2170
3. Igarashi T, Inatomi J, Sekine T,et al. Mutations in SLC4A4 cause permanent isolated procimal renal tubular acidosis with ocular abnormalities[J]. Nat Genet, 1999,23:264-266
4. Igarashi T, Inatomi J, Sekine T,et al.Novel non sense mutation in the Na+/HCO3-cotransporter gene(SLC4A4) in a patient with permanent isolated proximal renal tubular acidosis and bilateral glaucoma[J]. J Am Soc Nephrol,2001,12:713-718
5. Dinour D, Chang MH, Satoh J, et al. A novel missense mutation in the sodium bicarbonate cotransporter(NBCel/SLC4A4) causes proximal tubular acidosis and glaucoma through ion transport defects[J]. J Biol Chem,2004,279:52238-52246
6. Masashi Suzuki & Maria Helena Vaisbich & Hideomi Yamada, et al. Functional analysis of a novel missense NBC1 mutation and of other mutations causing proximal renal tubular acidosis[J]. Pflugers Arch - Eur J Physiol, 2008, 455:583-593
7. Donckerwolcke RA, Van Stekelenburg GJ, Tiddens HA. A case of bicarbonate-losing renal tubular acidosis with defective carboanhydrase activity [J]. Arch Dis Child, 1970,45:769-773
8. Sly WS, Hewett-Emmentt D, Whyte MP, Yu YS, et al. Carbonic anhydrase Ⅱ deficiency identified as the primary defect in the autosomal recessive syndrome of osteopetrosis with renal tubular acidosis and cerebral calcification[J]. Proc Natl Acad Sci USA, 1983,80:2752-2756
9. Venta PJ, Welty RJ, Johnson TM, et al. Carbonic anhydrase Ⅱ deficiency syndrome in a Belgian family is caused by a point mutation at an invariant histidine residue(107His*Try):complete structure of the normal human CAⅡgene[J].Am J Hum Gent,1991,49:1082-1090
10.Roth DE,Venta PJ,Tashian RE,et al.Molecular basis of human carbonic anhydrase Ⅱ deficiency[J].Proc Natl Acad Sci USA,1992,89:1804-1808
11.Karin E.Finberg,Carsten A.Wagner,Matthew A.Bailey,et al.The Bl-subunit of the H~+-ATPase is required for maximal urinary acidification[J].PNAS,2005,102(38):13616-1362
12.PAUL A.STEHBERGER,NICOLE SCHULZ,KARIN E.FINBERG,et al.Localization and Regulation of the ATP6VOA4(a4)Vacuolar H~+-ATPase Subunit Defective in an Inherited Form of Distal Renal Tubular Acidosis[J].J Am Soc Nephrol,2003,14:3027-3038
13.郑法雷.肾小管酸中毒的诊断与治疗[J].医师进修杂志,2003,26(5):1-3
14.Eriksson P,Denneberg T,Enestrom S,et al.Urolithoasis and distal renal tubular acidosis preceding primary Sjogren syndrome:a retrospective study 5-53 years after the presentation of urolithiasis[J].J Intern Med,1996,239:438-488
15.Lai LW,Chan DM,Erichson RP,et al.Correction of renal tubular acidosis in carbonic anyhdras Ⅱ-deficiant mice with gene therapy[J].J Clin Invest,1998,101:1305-1325
16.杨继红.肾小管酸中毒的临床表现与治疗[J].新医学,2007,38(9):570-572
17.张乃峥主编.临床风湿病学[M].上海:上海科学技术出版社,1999,287-298
18.Aasarqd K,Haga HJ,Berg KJ,et al.Renal involvement in primary Sjogren's syndrome[J].Q J Med,2000,93:297-304
19.董怡,张乃峥.干燥综合征的肾脏损害[J].中华内科杂志,1988,27:162-164
20.董德长主编.实用肾脏病学[M].上海:上海科学技术出版社,1999,1036-1045
21.Pertovaara M,Korpela M,Kouri T,et al.The occurrence of renal involvement in primary Sjogren's syndrome:a study of 78 patients[J].Rheumatology,1999,38:1113- 1120
22.Howard E.Corey·Alfredo Vallo'Juan Rodr_guez-Soriano.An analysis of renal tubular acidosis by the Stewart method[J].Pediatr Nephrol,2006,21:206-211
23.廖二元,莫朝晖主编.内分泌学..第2版[M].北京:人民卫生出版社,2007,601-604
24.杨华彬.Graves病并远端肾小管性酸中毒与周期性瘫痪[J].临床误诊误治,2004,17(12):905-906
25.钱桐荪.远端肾小管性酸中毒的病因和临床表现[J].新医学,2007,38(9):568-570
26.Fiona E.Karet,Karin E.Finberg,Raoul D.Nelson,et al.Mutations in the gene encoding B1 subunit of H~+-ATPase cause renal tubular acidosis with sensorineural deafness[J].Nature Genetics,1999,21(1):84-9027.Ya Su,Aiwu Zhou,Rafia S.Al-Lamki,et al.The a-subunit of the v-type H~+-AYPase interact with phosphofructokinase-1 in humans[J].The journal of biological chemistry,2003,278(22):20013-20018
28.Cremers C.W.,Di Pietro A.,et al.Mutations in the gene encoding B1 subunit of H~+-ATPase cause RTA with sensorineural deafness[J].Nature genetics,1999,21:84-90
29.Carsten A.Wagner,Karin E.Finberg,Sylvie Breton,et al.Renal vacuolar H~+-ATPase[J].Physiol rev.2004,84(4):1263-314
30.Karin E.Finberga,Carsten A.Wagnerb,Paul A.Stehbergerb,et al.Molecular cloning and characterization of Atp6v1b1,the murine vacuolar H~+-ATPase B1-subunit[J].Gene,2003,318:25-34
31.Karin E.Finberg,Carsten A.Wagner,Matthew A.Bailey,et al.The B1-subunit of the H~+- AYPase is required for maximal urinary acidification[J].PNAS,2005,102(38)13616-13621
32.Florina Rothenberger,Ana Velic,Paul A.Stehberger,et al.Angiotensin Ⅱstimulates vacuolar H+ -ATPase activity in renal acid-secretory intercalated cells from the outer medullary collecting duct[J].J Am Soc Nephrol,2007,18:2085-2093
33.G Li,Q Yang,S Krishnan,et al.A novel cellular survival factor - the B2 subunit of vacuolar H~+-ATPase inhibits apoptosis[J].Cell death and differentiation,2006,13:2109-2117
34.陈慰峰主编.医学免疫学..第三版[M].北京:人民卫生出版社,2002,209-210
35.Fumi Takemoto,Junichi Hoshino,Naoki Sawa,et al.Autoantibodies against carbonic anhydrase Ⅱ are increased in renal tubular acidosis associated with Sjogren syndrome[J].The American Journal of Medicine,2005,118:181-184
36.Kohriyama K,Katayama Y,Tsurusako Y.Relationship between primary Sjogren's syndrome and autoimmune thyroid disease[L].Nippon Rinsho,1999,57:1878-1881
37.任红,陈楠,陈晓农,等.干燥综合征合并肾脏损害147例临床病理及随访情况[J].中华风湿病学杂志,2005,9(6):351-353
38.周叶青,胡曼东.舍格伦综合症-合并肾小管酸中毒的免疫学研究[J].口腔医学,2001,21(4):197-198
39.江永娣,陈楠,王朝晖,等.肾小管性酸中毒115例临床分析[J].中华内分泌代谢杂志,1998,14:155-158
40.K.W JOO,U,S JEON J S HAN,S KIM.JS.LEE,et al.Absence of H~+-ATPase in the intercalated cells of renal tissues in classic distal renal tubular acidosis[J].Clinical nephrology,1998,149(4):226-231
41.JIN SUK HAN,GHEUN-HO KIM,JIN KIM,et al.Secretory-Defect Distal Renal Tubular Acidosis Is Associatedwith Transporter Defect in H~+-AYPase and AnionExchanger-1[J].J Am Soc Nephrol,2002,13:1425-1432