CAJ | 학술논문

目的探讨熊去氧胆酸(UDCA)对α-萘异硫氰酸酯(ANIT)诱导大鼠胆汁淤积性肝损伤的保护作用及作用机制.方法取SD大鼠48只,其中42只以ANIT 100 mg/kg灌胃造成急性肝损伤,在肝损伤后24 h处死6只,其余均分为对照组和UDCA组各18只.对照组予0.9％NaCl溶液灌胃,UDCA组予20 mg/kg UDCA灌胃,分别在造模后48、72、96 h各处死6只,另6只不予处理为空白对照组.所有大鼠处死留取血清和肝组织,检测血清ALT、AST、Tbil、总胆汁酸(TBA),ELISA法检测血清IL-10、IL-6、TNF-α,实时荧光定量PCR检测肝组织多药耐药相关蛋白2(Mrp2) mRNA,HE染色观察肝组织炎性反应活动度.结果肝损伤造模后48 h,UDCA组和对照组血清Tbil[(143.80±12.08) μmol/L比(178.50±15.19)μmol/L]、TBA[( 13.15±3.81) μmol/L比(21.68±7.93)μmol/L]、IL-10[(44.13±3.68)ng/L比(37.15±6.25)ng/L]、IL-6[(50.80±2.09) ng/L比(57.32±4.63)ng/L]、TNF-α[(17.53±0.84)ng/L比(19.10±1.64) ng/L]比较,差异均有统计学意义(P＜0.01或P＜0.05)；肝损伤造模后72 h,UDCA组和对照组血清ALT[(721.67±97.54)U/L比(929.50±148.29)U/L]、IL-10[(54.68±6.79)ng/L比(43.85±4.08)ng/L]比较,差异均有统计学意义(P＜0.01或P＜0.05)；肝损伤造模后96 h,UDCA组和对照组血清ALT[(156.83±14.99)U/L比(250.67±42.29)U/L]、AST[(143.67±27.45) U/L比(206.00±63.94)U/L]、Tbil[(23.53±5.08)μmol/L比(34.02±9.98)μmol/L]比较,差异有统计学意义(P＜0.01或P＜0.05).肝损伤造模后UDCA组和对照组肝组织Mrp2 mRNA表达在48 h(0.77±0.21、0.46±0.25)、72 h(2.27±0.84、1.10±0.38)、96 h(3.64±0.54、2.75±0.69)各时间点差异均有统计学意义(P＜0.01或P＜0.05).结论 UDCA对ANIT诱导的胆汁淤积性肝损伤保护作用机制可能和调控血清细胞因子及肝脏Mrp2表达有关.
목적 탐토웅거양담산(UDCA)대α-내이류청산지(ANIT)유도대서담즙어적성간손상적보호작용급작용궤제.방법 취SD대서48지,기중42지이ANIT 100 mg/kg관위조성급성간손상,재간손상후24 h처사6지,기여균분위대조조화UDCA조각18지.대조조여0.9％NaCl용액관위,UDCA조여20 mg/kg UDCA관위,분별재조모후48、72、96 h각처사6지,령6지불여처리위공백대조조.소유대서처사류취혈청화간조직,검측혈청ALT、AST、Tbil、총담즙산(TBA),ELISA법검측혈청IL-10、IL-6、TNF-α,실시형광정량PCR검측간조직다약내약상관단백2(Mrp2) mRNA,HE염색관찰간조직염성반응활동도.결과 간손상조모후48 h,UDCA조화대조조혈청Tbil[(143.80±12.08) μmol/L비(178.50±15.19)μmol/L]、TBA[( 13.15±3.81) μmol/L비(21.68±7.93)μmol/L]、IL-10[(44.13±3.68)ng/L비(37.15±6.25)ng/L]、IL-6[(50.80±2.09) ng/L비(57.32±4.63)ng/L]、TNF-α[(17.53±0.84)ng/L비(19.10±1.64) ng/L]비교,차이균유통계학의의(P＜0.01혹P＜0.05)；간손상조모후72 h,UDCA조화대조조혈청ALT[(721.67±97.54)U/L비(929.50±148.29)U/L]、IL-10[(54.68±6.79)ng/L비(43.85±4.08)ng/L]비교,차이균유통계학의의(P＜0.01혹P＜0.05)；간손상조모후96 h,UDCA조화대조조혈청ALT[(156.83±14.99)U/L비(250.67±42.29)U/L]、AST[(143.67±27.45) U/L비(206.00±63.94)U/L]、Tbil[(23.53±5.08)μmol/L비(34.02±9.98)μmol/L]비교,차이유통계학의의(P＜0.01혹P＜0.05).간손상조모후UDCA조화대조조간조직Mrp2 mRNA표체재48 h(0.77±0.21、0.46±0.25)、72 h(2.27±0.84、1.10±0.38)、96 h(3.64±0.54、2.75±0.69)각시간점차이균유통계학의의(P＜0.01혹P＜0.05).결론 UDCA대ANIT유도적담즙어적성간손상보호작용궤제가능화조공혈청세포인자급간장Mrp2표체유관.
Objective To investigate the protective effects and mechanism of ursodeoxycholic acid (UDCA) on α-naphthylisothi (ANIT)-induced cholestatic liver injury in rats.Methods A total of 48 Sprague-Dawley (SD) rats were selected.Fouty-two rats were gavaged with ANIT (100 mg/kg) to induce acute liver injury,six rats were sacrificed 24 hours after the liver injury and the rats left were evenly divided into control group which were gavaged with saline and UDCA group which were gavaged with UDCA (20 mg/kg).Six rats were sacrificed at 48 hours,72 hours and 96 hours after modeling.The six untreated rats were set as blank control group.Serum and liver tissues of all rats were kept after sacrificed.Serum levels of alanine transaminase (ALT),aspartate transaminase (AST),total bilirubin (TBil) and total bile acid (TBA) were tested,interleukin-10 (IL-10),interleukin-6 (IL-6),and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA).The expression of multidrug resistance associated protein2 (Mrp2) at mRNA level in liver tissue was measured by quantitative real-time polymerase chain reaction (qRT-PCR) and the inflammatory reaction activity of liver tissues was inspected with Haematoidin-Eosin (HE) staining under microscope.Results At 48 hours after liver injury modeling,serum TBil (143.80± 12.08) μmol/L vs.(178.50±15.19) μmol/L,TBA (13.15±3.81) μmol/L vs.(21.68±7.93)mol/L,IL-10 (44.13±3.68,37.15±6.25 ng/L),IL-6(50.80±2.09,57.32±4.63 ng/L) and TNF-α (17.53±0.84) ng/L vs,(19.10±1.64) ng/L of UDCA group and control group were compared,and the differences were statistically significant (P ＜ 0.01 or P＜ 0.05).At 72 hours after liver injury modeling,serum ALT (721.67±97.54) U/L vs.(929.50±148.29) U/L and IL-10 (54.68±6.79)ng/L vs.(43.85±4.08) ng/L of UDCA group and control group were compared,and the differences were statistically significant (P＜0.01 or P＜0.05).At 96 hours after liver injury modeling,serum ALT (156.83±14.99) U/L vs.(250.67±42.29) U/L,AST (143.67±27.45) U/L vs.(206.00±63.94) U/L and TBil (23.53±5.08) μmol/L vs.(34.02±9.98) μmol/L of UDCA group and control group were compared,and the differences were statistically significant (P＜0.01 or P＜0.05).The differences of Mrp2 expression at mRNA level in liver tissues between UDCA group and control group at 48 hours (0.77 ± 0.21,0.46 ± 0.25),72 hours (2.27 ±0.84,1.10 ±0.38) and 96 hours (3.64±0.54,2.75±0.69) after liver injury modeling were statistically significant (P＜0.01 or P＜0.05).Conclusion The mechanism of the protective effects of UDCA on ANIT-induced liver injury may be related with the regulation of serum cytokines and liver Mrp2 expression.