中华妇产科杂志
中華婦產科雜誌
중화부산과잡지
CHINESE JOUNAL OF OBSTETRICS AND GYNECOLOGY
2010年
6期
440-444
,共5页
时青云%林宇庚%周新%林颖奇%闫时
時青雲%林宇庚%週新%林穎奇%閆時
시청운%림우경%주신%림영기%염시
胆汁淤积,肝内%妊娠并发症%PPARα%甾类羟化酶类%雌激素受体α%雌激素受体β
膽汁淤積,肝內%妊娠併髮癥%PPARα%甾類羥化酶類%雌激素受體α%雌激素受體β
담즙어적,간내%임신병발증%PPARα%치류간화매류%자격소수체α%자격소수체β
Cholestasis,intrahepatic%Pregnancy complications%PPAR alpha%Steroid hydroxylases%Estrogen receptor alpha%Estrogen receptor beta
目的 探讨过氧化物酶增殖体激活受体α(PPARα)、氧固醇7α羟化酶(CYP7B1)、雌激素受体(ER)α及ERβ之间的调控关系与孕鼠肝内胆汁淤积发生的相关性.方法 选择清洁级SD孕鼠80只,随机分为4组,每组20只,自孕第13天起:对照组孕鼠皮下注射精制植物油2.0ml·kg-1·d-1;低剂量组孕鼠皮下注射17-α-乙炔雌二醇(1.0 mg·kg-1·d-1);中剂量组孕鼠皮下注射17-α-乙炔雌二醇(1.25 mg·lg-1·d-1);高剂量组孕鼠皮下注射17-α-乙炔雌二醇(1.5 mg·kg-1·d-1).4组孕鼠于妊娠第21天处死后提取肝脏组织.应用酶联免疫吸附试验检测各组孕鼠血清中丙氨酸转氨酶(ALT)、门冬氨酸转氨酶(AST)、总胆酸(TBA)及胆红素(BIL)水平;应用实时定量PCR技术检测各组孕鼠肝脏组织中PPARα mRNA、CYP7B1 mRNA、ERα mRNA及ERβ mRNA的表达水平.结果 (1)生化指标:对照组孕鼠ALT、AST、TBA及BIL水平分别为(41.1±2.8)U/L、(44.4±3.6)U/L、(26.4±5.6)μmol/L、(2.8±0.2)U/L,低剂量组孕鼠分别为(48.2±3.4)U/L、(47.9±3.7)U/L、(36.4±4.2)μmol/L、(4.2±0.2)U/L,中剂量组孕鼠分别为(70.4±5.3)U/L、(68.4±5.6)U/L、(64.3±3.8)μmol/L、(6.2±1.2)U/L,高剂量组孕鼠分别为(72.4±7.6)U/L、(70.2±3.8)U/L、(72.4±7.8)μmol/L、(8.2±2.2)U/L.低剂量组、中剂量组、高剂量组孕鼠ALT、AST、TBA、BIL水平明显高于对照组(P<0.05);中剂量组、高剂量组孕鼠各生化指标水平明显高于低剂量组(P<0.05).(2)ERαmRNA及ERβmRNA表达水平:ERαmRNA的表达水平在低剂量组(0.76±0.02)、中剂量组(0.99±0.04)和高剂量组(1.21±0.01)孕鼠肝脏组织中呈逐渐升高趋势(P<0.05),并明显高于对照组(0.65±0.01),分别与对照组比较,差异均有统计学意义(P<0.05);ERβ表达水平在4组孕鼠间分别比较,差异均无统计学意义(P>0.05).(3)CYP7B1 mRNA及PPARα mRNA表达水平:CYP7B1 mRNA的表达水平在低剂量组(0.93±0.01)、中剂量组(0.99±0.06)和高剂量组(1.22±0.04)孕鼠肝脏组织中呈逐渐升高趋势(P<0.05),并明显高于对照组(0.75±0.02),分别与对照组比较,差异均有统计学意义(P<0.05);PPARα mRNA表达水平在低剂量组(0.83±0.05)、中剂量组(0.71±0.02)和高剂量组(0.64±0.03)孕鼠肝脏组织中呈逐渐降低趋势(P<0.05),并明显低于对照组(1.35±0.05),分别与对照组比较,差异均有统计学意义(P<0.05).结论 随着雌激素剂量的增加,PPARα表达水平降低,对CYP7B1表达的抑制作用解除,而导致CYP7B1表达水平升高;而CYP7B1有促进ERα高表达的作用,最终由ERα介导了雌激素诱导的肝内胆汁淤积的发生.提示PPARα、CYP7B1及ER的异常表达,是调控雌激素诱导孕鼠肝内胆汁淤积的发生机制之一.
目的 探討過氧化物酶增殖體激活受體α(PPARα)、氧固醇7α羥化酶(CYP7B1)、雌激素受體(ER)α及ERβ之間的調控關繫與孕鼠肝內膽汁淤積髮生的相關性.方法 選擇清潔級SD孕鼠80隻,隨機分為4組,每組20隻,自孕第13天起:對照組孕鼠皮下註射精製植物油2.0ml·kg-1·d-1;低劑量組孕鼠皮下註射17-α-乙炔雌二醇(1.0 mg·kg-1·d-1);中劑量組孕鼠皮下註射17-α-乙炔雌二醇(1.25 mg·lg-1·d-1);高劑量組孕鼠皮下註射17-α-乙炔雌二醇(1.5 mg·kg-1·d-1).4組孕鼠于妊娠第21天處死後提取肝髒組織.應用酶聯免疫吸附試驗檢測各組孕鼠血清中丙氨痠轉氨酶(ALT)、門鼕氨痠轉氨酶(AST)、總膽痠(TBA)及膽紅素(BIL)水平;應用實時定量PCR技術檢測各組孕鼠肝髒組織中PPARα mRNA、CYP7B1 mRNA、ERα mRNA及ERβ mRNA的錶達水平.結果 (1)生化指標:對照組孕鼠ALT、AST、TBA及BIL水平分彆為(41.1±2.8)U/L、(44.4±3.6)U/L、(26.4±5.6)μmol/L、(2.8±0.2)U/L,低劑量組孕鼠分彆為(48.2±3.4)U/L、(47.9±3.7)U/L、(36.4±4.2)μmol/L、(4.2±0.2)U/L,中劑量組孕鼠分彆為(70.4±5.3)U/L、(68.4±5.6)U/L、(64.3±3.8)μmol/L、(6.2±1.2)U/L,高劑量組孕鼠分彆為(72.4±7.6)U/L、(70.2±3.8)U/L、(72.4±7.8)μmol/L、(8.2±2.2)U/L.低劑量組、中劑量組、高劑量組孕鼠ALT、AST、TBA、BIL水平明顯高于對照組(P<0.05);中劑量組、高劑量組孕鼠各生化指標水平明顯高于低劑量組(P<0.05).(2)ERαmRNA及ERβmRNA錶達水平:ERαmRNA的錶達水平在低劑量組(0.76±0.02)、中劑量組(0.99±0.04)和高劑量組(1.21±0.01)孕鼠肝髒組織中呈逐漸升高趨勢(P<0.05),併明顯高于對照組(0.65±0.01),分彆與對照組比較,差異均有統計學意義(P<0.05);ERβ錶達水平在4組孕鼠間分彆比較,差異均無統計學意義(P>0.05).(3)CYP7B1 mRNA及PPARα mRNA錶達水平:CYP7B1 mRNA的錶達水平在低劑量組(0.93±0.01)、中劑量組(0.99±0.06)和高劑量組(1.22±0.04)孕鼠肝髒組織中呈逐漸升高趨勢(P<0.05),併明顯高于對照組(0.75±0.02),分彆與對照組比較,差異均有統計學意義(P<0.05);PPARα mRNA錶達水平在低劑量組(0.83±0.05)、中劑量組(0.71±0.02)和高劑量組(0.64±0.03)孕鼠肝髒組織中呈逐漸降低趨勢(P<0.05),併明顯低于對照組(1.35±0.05),分彆與對照組比較,差異均有統計學意義(P<0.05).結論 隨著雌激素劑量的增加,PPARα錶達水平降低,對CYP7B1錶達的抑製作用解除,而導緻CYP7B1錶達水平升高;而CYP7B1有促進ERα高錶達的作用,最終由ERα介導瞭雌激素誘導的肝內膽汁淤積的髮生.提示PPARα、CYP7B1及ER的異常錶達,是調控雌激素誘導孕鼠肝內膽汁淤積的髮生機製之一.
목적 탐토과양화물매증식체격활수체α(PPARα)、양고순7α간화매(CYP7B1)、자격소수체(ER)α급ERβ지간적조공관계여잉서간내담즙어적발생적상관성.방법 선택청길급SD잉서80지,수궤분위4조,매조20지,자잉제13천기:대조조잉서피하주사정제식물유2.0ml·kg-1·d-1;저제량조잉서피하주사17-α-을결자이순(1.0 mg·kg-1·d-1);중제량조잉서피하주사17-α-을결자이순(1.25 mg·lg-1·d-1);고제량조잉서피하주사17-α-을결자이순(1.5 mg·kg-1·d-1).4조잉서우임신제21천처사후제취간장조직.응용매련면역흡부시험검측각조잉서혈청중병안산전안매(ALT)、문동안산전안매(AST)、총담산(TBA)급담홍소(BIL)수평;응용실시정량PCR기술검측각조잉서간장조직중PPARα mRNA、CYP7B1 mRNA、ERα mRNA급ERβ mRNA적표체수평.결과 (1)생화지표:대조조잉서ALT、AST、TBA급BIL수평분별위(41.1±2.8)U/L、(44.4±3.6)U/L、(26.4±5.6)μmol/L、(2.8±0.2)U/L,저제량조잉서분별위(48.2±3.4)U/L、(47.9±3.7)U/L、(36.4±4.2)μmol/L、(4.2±0.2)U/L,중제량조잉서분별위(70.4±5.3)U/L、(68.4±5.6)U/L、(64.3±3.8)μmol/L、(6.2±1.2)U/L,고제량조잉서분별위(72.4±7.6)U/L、(70.2±3.8)U/L、(72.4±7.8)μmol/L、(8.2±2.2)U/L.저제량조、중제량조、고제량조잉서ALT、AST、TBA、BIL수평명현고우대조조(P<0.05);중제량조、고제량조잉서각생화지표수평명현고우저제량조(P<0.05).(2)ERαmRNA급ERβmRNA표체수평:ERαmRNA적표체수평재저제량조(0.76±0.02)、중제량조(0.99±0.04)화고제량조(1.21±0.01)잉서간장조직중정축점승고추세(P<0.05),병명현고우대조조(0.65±0.01),분별여대조조비교,차이균유통계학의의(P<0.05);ERβ표체수평재4조잉서간분별비교,차이균무통계학의의(P>0.05).(3)CYP7B1 mRNA급PPARα mRNA표체수평:CYP7B1 mRNA적표체수평재저제량조(0.93±0.01)、중제량조(0.99±0.06)화고제량조(1.22±0.04)잉서간장조직중정축점승고추세(P<0.05),병명현고우대조조(0.75±0.02),분별여대조조비교,차이균유통계학의의(P<0.05);PPARα mRNA표체수평재저제량조(0.83±0.05)、중제량조(0.71±0.02)화고제량조(0.64±0.03)잉서간장조직중정축점강저추세(P<0.05),병명현저우대조조(1.35±0.05),분별여대조조비교,차이균유통계학의의(P<0.05).결론 수착자격소제량적증가,PPARα표체수평강저,대CYP7B1표체적억제작용해제,이도치CYP7B1표체수평승고;이CYP7B1유촉진ERα고표체적작용,최종유ERα개도료자격소유도적간내담즙어적적발생.제시PPARα、CYP7B1급ER적이상표체,시조공자격소유도잉서간내담즙어적적발생궤제지일.
Objective To investigate the relationship between interaction of peroxisome proliferators-activated receptor alpha (PPARα), cytochrome P450 oxysterol 7α-hydroxylase (CYP7B1) and estrogen receptor (ER) and intrahepatic cholestasis in pregnant rats. Methods Eighty clean SD pregnant rats were selected and divided into four groups randomly with 20 in each. Since the 13th day of pregnancy,rats in the control group was injected subcutaneously with refined vegetable oil 2.0 ml · kg-1 · d -1 , those in the low-dose, moderate-dose and high-dose groups received 17-α-ethynylestradiol (EE) 1.0 mg · kg-1 · d-1,1.25 mg · kg-1 · d-1 and 1.5 mg · kg-1 · d-1, respectively. All rats were sacrificed at the 21at day of pregnancy and maternal hepatic tissues were collected. The serum levels of alanine aminotransferase(ALT), aspartate transaminase (AST), total bile acid (TBA) and bilirubin (BIL) were determined by enzyme linked immunosorbent assay (ELISA). The mRNA expressions of PPARα, CYP7B1, Erα and Erβ in maternal rat livers were examined by real-time PCR. Results (1) Biochemical indicators: the serum levels of ALT,AST, TBA and BIL were significantly lower in the control group than in the rest 3 groups,respectively [ control group: (41.1 ± 2.8 ) U/L, (44.4 ± 3.6) U/L, (26.4 ± 5.6 ) μmol/L and( 2.8 ± 0.2)U/L;low-dose group: (48.2 ±3.4) U/L,(47.9 ±3.7) U/L,(36.4 ±4.2) μmol/L and (4.2 ±0.2) U/L;moderate-dose group: (70.4 ± 5.3 ) U/L, (68.4 ± 5.6) U/L, (64.3 ± 3.8 ) μmol/L and ( 6.2 ± 1.2)U/L; high-dose group: (72.4 ±7.6) U/L, (70.2 ±3.8) U/L, (72.4 ±7.8) μmol/L and (8.2 ±2.2)U/L, P<0.05], and those in the moderate or high-dose groups were higher than in the low-dose group (P<0.05). (2) mRNA expression of Erα and Erβ: the mRNA expression of Erα in pregnant rat livers increased in a dose-dependent manner, which were all significantly higher than that in the control group,respectively ( low-dose group: 0.76 ± 0.02 ); moderate -dose group: ( 0.99 ± 0.04; high-dose group:1.21 ±0.01 ;control group:0.65 ±0.01, P <0.05), but no difference was found among the 4 groups in the mRNA expression of Erβ ( P > 0.05 ). (3) mRNA expression of CYP7B1 and PPARα: the mRNA expression of CYP7B1 in pregnant rat livers increased from the low-dose group to the high-dose group, and were all higher than that of the control group ( low-dose group: 0.93 ± 0.01; moderate-dose group: 0.99 ±0.06; high-dose group: 1.22 ± 0.04; control group: 0.75 ± 0.02, P < 0.05 ). However, the mRNA expression of PPARα decreased from the low-dose group to the high-dose group, and were all lower than that of the control group (low-dose group: 0.83 ± 0.05; moderate-dose group: 0.71 ± 0.02; high-dose group:0.64 ± 0.03; control group: 1.35 ± 0. 05; P < 0.05 ) . Conclusions The down regulated mRNA expression of PPARα, caused by higher dose of estrogen, may increase the expression of CYP7B1 due to the ineffectiveness of the inhibition of PPARα on CYP7B1, which may further stimulate the Erα activity and then induce intrahepatic cholestasis. Abnormal expression of PPARα, CYP7B1 and ER may play a role in the pathogenesis of estrogen-induced intrahepatic cholestasis.
