中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2009年
12期
925-929
,共5页
乔颖进%谌贻璞%芮宏亮%董鸿瑞%刘章锁
喬穎進%諶貽璞%芮宏亮%董鴻瑞%劉章鎖
교영진%심이박%예굉량%동홍서%류장쇄
马兜铃酸%贫血%红细胞生成素%肾小管周围毛细血管%低氧诱导因子
馬兜鈴痠%貧血%紅細胞生成素%腎小管週圍毛細血管%低氧誘導因子
마두령산%빈혈%홍세포생성소%신소관주위모세혈관%저양유도인자
Aristolochic acid%Anemia%Erythropoietin%Peritubular capillary%Hypoxia inducible factors
目的 探讨慢性马兜铃酸肾病(CAAN) 贫血的发牛机制.方法 用62只SD大鼠筛查血红蛋白(Hb),求正常值.随机选择24只Hb正常的大鼠,分为对照组及模型组.各12只,后者予关木通浸膏水溶液fRJ断灌胃制作CAAN模型.在给药前及8周末,分别检测两组各6只大鼠体质量、Hb、尿蛋白量(24 h)及肌酐清除率(Ccr),然后处死大鼠,留取肾组织做 Masson染色观察肾间质纤维化程度;实时定量PCR法检测肾组织中红细胞生成素(EPO) mRNA表达;免疫组化染色观察肾组织中I型胶原(Col I)、氨基肽酶P(APP)、低氧诱导因子(HIF)lα及2α的蛋白表达.结果 大鼠的正常Hb值为(155.9±16.5)g/L,低于123.6 g/L为贫血.给药前,对照组与模型组间各指标的差异均无统计学意义.8周末时,与对照组比较,模型组大鼠Hb和Ccr显著下降(121.66±15.68)g/L比(169.00±12.89)g/L,(0.63±0.13)ml/min 比(1.27±0.18)ml/minl;尿蛋白量(24 h)显著增多l(27.04±9.40)mg/d比(6.11±0.84)mg/d];肾间质纤维化相对面积及Coll相对面积显著增加[(12.89±2.33)%比(0.55±0.10)%,(13.92± 2.92)%比(1.32±0.84)%];肾组织APP蛋白表达和EPO mRNA表达显著下调[(0.55±0.23)% 比(3.77±1.06)%.0.005±0.001比0.032±0.013];HIF-1α和 HIF-2α蛋白表达显著上调(2.55±0.16比1.12±0.46,2.33±0.33比1.15±0.27)(均P<0.01).结论 CAAN的贫血发生可能与肾小管周毛细血管毁坏所导致的EPO产生减少有关,虽然HIF表达已代偿性增强,但仍未能 阻止贫血发生.
目的 探討慢性馬兜鈴痠腎病(CAAN) 貧血的髮牛機製.方法 用62隻SD大鼠篩查血紅蛋白(Hb),求正常值.隨機選擇24隻Hb正常的大鼠,分為對照組及模型組.各12隻,後者予關木通浸膏水溶液fRJ斷灌胃製作CAAN模型.在給藥前及8週末,分彆檢測兩組各6隻大鼠體質量、Hb、尿蛋白量(24 h)及肌酐清除率(Ccr),然後處死大鼠,留取腎組織做 Masson染色觀察腎間質纖維化程度;實時定量PCR法檢測腎組織中紅細胞生成素(EPO) mRNA錶達;免疫組化染色觀察腎組織中I型膠原(Col I)、氨基肽酶P(APP)、低氧誘導因子(HIF)lα及2α的蛋白錶達.結果 大鼠的正常Hb值為(155.9±16.5)g/L,低于123.6 g/L為貧血.給藥前,對照組與模型組間各指標的差異均無統計學意義.8週末時,與對照組比較,模型組大鼠Hb和Ccr顯著下降(121.66±15.68)g/L比(169.00±12.89)g/L,(0.63±0.13)ml/min 比(1.27±0.18)ml/minl;尿蛋白量(24 h)顯著增多l(27.04±9.40)mg/d比(6.11±0.84)mg/d];腎間質纖維化相對麵積及Coll相對麵積顯著增加[(12.89±2.33)%比(0.55±0.10)%,(13.92± 2.92)%比(1.32±0.84)%];腎組織APP蛋白錶達和EPO mRNA錶達顯著下調[(0.55±0.23)% 比(3.77±1.06)%.0.005±0.001比0.032±0.013];HIF-1α和 HIF-2α蛋白錶達顯著上調(2.55±0.16比1.12±0.46,2.33±0.33比1.15±0.27)(均P<0.01).結論 CAAN的貧血髮生可能與腎小管週毛細血管燬壞所導緻的EPO產生減少有關,雖然HIF錶達已代償性增彊,但仍未能 阻止貧血髮生.
목적 탐토만성마두령산신병(CAAN) 빈혈적발우궤제.방법 용62지SD대서사사혈홍단백(Hb),구정상치.수궤선택24지Hb정상적대서,분위대조조급모형조.각12지,후자여관목통침고수용액fRJ단관위제작CAAN모형.재급약전급8주말,분별검측량조각6지대서체질량、Hb、뇨단백량(24 h)급기항청제솔(Ccr),연후처사대서,류취신조직주 Masson염색관찰신간질섬유화정도;실시정량PCR법검측신조직중홍세포생성소(EPO) mRNA표체;면역조화염색관찰신조직중I형효원(Col I)、안기태매P(APP)、저양유도인자(HIF)lα급2α적단백표체.결과 대서적정상Hb치위(155.9±16.5)g/L,저우123.6 g/L위빈혈.급약전,대조조여모형조간각지표적차이균무통계학의의.8주말시,여대조조비교,모형조대서Hb화Ccr현저하강(121.66±15.68)g/L비(169.00±12.89)g/L,(0.63±0.13)ml/min 비(1.27±0.18)ml/minl;뇨단백량(24 h)현저증다l(27.04±9.40)mg/d비(6.11±0.84)mg/d];신간질섬유화상대면적급Coll상대면적현저증가[(12.89±2.33)%비(0.55±0.10)%,(13.92± 2.92)%비(1.32±0.84)%];신조직APP단백표체화EPO mRNA표체현저하조[(0.55±0.23)% 비(3.77±1.06)%.0.005±0.001비0.032±0.013];HIF-1α화 HIF-2α단백표체현저상조(2.55±0.16비1.12±0.46,2.33±0.33비1.15±0.27)(균P<0.01).결론 CAAN적빈혈발생가능여신소관주모세혈관훼배소도치적EPO산생감소유관,수연HIF표체이대상성증강,단잉미능 조지빈혈발생.
Objective To study the pathogenesis of anemia in chronic aristolochic acid nephmpathy(CAAN) rats. Methods The hemoglobin(Hb)values of sixty-two male SD rats were assayed to determine its normal range.Among them,24 rats with normal Hb value were randomly divided into 2 groups:model group (MG)in which rats received the extract of Aristololochia manshuriensis Kom (AmK) by gavage,and control group (CG) received tap water only by gavage.Body weisht(BW),Hb,24 h urinary protein excretion(UP)and creatinine clearance (Ccr)of 6 rats in each group were measured before administration and at the end of the 8th week, respeetively.then these rats were sacrificed.The relative area of renal interstitial fibrosis was measured by microscopy.The mRNA expression of erythropoietin (EPO)in kidney tissue Was determined by real-time RT-PCR;protein expression of type I collagen(Coll),aminopeptidase P (APP),hypoxia indHeible factor let and 2α(HIF-1α and HIF-2α)in kidney tissue Was examined by immunohistochemistry staining. Results Hb values of normal rats presented normal distribution. The normal Hb was (155.9±16.5) g/L. Rat anemia was diagnosed when Hb was below 123.6 g/L. There was no difference in all the examination results between CG and MG before administration (P>0.05). Compared with CG, the Hb and Cer in MG were significantly decreased [(121.66±15.68) g/L vs (169.00±12.89) g/L, (0.63±0.13) ml/min vs (1.27±0.18) ml/min, P< 0.01], and the UP in MG was significantly increased at the end of the 8th week [(27.04±9.40) mg/d vs (6.11±0.84) mg/d, P<0.01]; the relative areas of fibrosis and Col l in renal interstitium of MG were significantly enlarged [(12.89±2.33)% vs (0.55±0.10)%, (13.92±2.92)% vs (1.32±0.84)%, P<0.01]; the protein expression of APP and the mRNA expression of EPO in the kidney tissue of MG were significantly down-regulated [(0.55±0.23)% vs (3.77±1.06)%, 0.005±0.001 vs 0.032±0.013, P<0.01]; the protein expression of HIF-lα and HIF-2α in the kidney tissue of MG was significantly up-regulated (2.55±0.16 vs 1.12±0.46, 2.33±0.33 vs 1.15±0.27, P<0.01), at the end of the 8th week. Conclusions The pathogenesis of anemia in CAAN may be due to the decreased production of EPO caused by the destruction of peritubular capillary. The compensatory up-regulation of HIF-lα and HIF-2α expression can not prevent the anemia development.
