目的 探討膽淤方對(duì)1，4-二氫-2，4，6-三甲基-3，5-吡啶二甲酸二乙酯（DDC）誘導(dǎo)膽汁淤積小鼠血清膽汁酸代謝的影響。方法 40只雄性C57BL/6J小鼠隨機(jī)分為4組：對(duì)照（N）組、DDC模型（M）組、奧貝膽酸（O）組、膽淤方（D）組，每組10只。N組小鼠用普通飼料喂養(yǎng)，M、O和D組小鼠用DDC飼料喂養(yǎng)，連續(xù)6周。造模第3周開始藥物干預(yù)，O組ig予奧貝膽酸溶液（0.03 g·kg^-1），D組ig予膽淤方藥液（33 g·kg^-1），M組和N組予相同體積的雙蒸水。干預(yù)4周，收集小鼠血清和肝組織，試劑盒檢測(cè)血清丙氨酸氨基轉(zhuǎn)移酶（ALT）、天冬氨酸氨基轉(zhuǎn)移酶（AST）、堿性磷酸酶（ALP）、總膽紅素（TBIL）和總膽汁酸（TBA）水平，HE染色觀察肝組織炎癥反應(yīng)，CK19染色觀察細(xì)膽管增生反應(yīng)，天狼腥紅染色觀察纖維化，采用液質(zhì)聯(lián)用技術(shù)檢測(cè)血清膽汁酸譜。結(jié)果 O組和D組ALT、AST、ALP、TBA、TBIL水平均較M組顯著下降（P＜0.05）；D組ALT、AST水平較O組顯著下降（P＜0.05）。D組小鼠肝組織炎癥反應(yīng)、纖維化及細(xì)膽管增生較M組減輕。D組血清初級(jí)膽汁酸膽酸（CA）和ω-鼠膽酸（ω-MCA）水平均較M組顯著下降（P＜0.001）；D組血清次級(jí)膽汁酸甘氨膽酸（GCA）、?；悄懰幔═CA）、?；切苋パ跄懰幔═UDCA）、?；蛆Z去氧膽酸（TCDCA）、?；侨パ跄懰幔═DCA）、?；秦i去氧膽酸（THDCA）較M組顯著下降（P＜0.05、0.01、0.001），O組的CA、ω-MCA、TCDCA、GCA和TCA水平較M組顯著下降（P＜0.001），D組血清TUDCA水平較O組下降更加顯著（P＜0.05）。熱圖分析顯示13種膽汁酸可將M與N組、O與M組、D與M組、D與O組血清膽汁酸譜兩兩區(qū)分。結(jié)論 膽淤方可改善肝內(nèi)炎癥反應(yīng)，具有廣泛的降膽汁酸作用。

Objective To investigate the effect of Danyu Recipe on the serum bile acid metabolism in mice with cholestasis induced by DDC. Methods 40 male C57BL/6J mice were randomly divided into four groups, 10 normal group (N), 10 model group (M), 10 obeticholic acid group (O), and 10 Danyu Recipe group (D). The mice in group N were fed with normal feed. Mice in the M, O and D groups were fed with DDC feed. Modeling was continued for six weeks. Drug intervention was started in the 3rd week of modeling. The O group was orally administered with Obeticholic acid 0.03 g·kg^-1, and the D group was orally administered with Danyu decoction 33 g·kg^-1. The mice in group M and group N were given the same volume of double distilled water by gavage. Mice were treated after four weeks of intervention, and mouse serum and liver tissue were collected. The serum bile acid profile of four groups was detected by liquid chromatography-mass spectrometry. Results The levels of ALT, AST, ALP, TBA and TBIL in O group and D group were significantly lower than those in M group (P < 0.05). The levels of ALT and AST in group D were significantly lower than those in group O (P < 0.05). There was no significant difference in the levels of ALP, TBA and TBIL in group D and group O. The intrahepatic inflammatory reaction, fibrosis and hyperplasia of small bile ducts in group D were less than those in group M. Serum primary bile acid CA and ω-MCA levels in group D were significantly lower than those in group M (P < 0.05). The levels of serum secondary bile acids GCA, TCA, TUDCA, TCDCA, TDCA, and THDCA in group D were significantly lower than those in group M (P < 0.05). The levels of GCA and TCA in group O were significantly lower than those in group M (P < 0.05). However, the level of serum TUDCA in group D was more significantly lower than that in group O (P < 0.05). Heat map analysis showed that 13 kinds of bile acids could distinguish the serum bile acid profiles of M and N groups, O and M groups, D and M groups, and D and O groups. Conclusion Danyu Recipe can improve the inflammatory response in the liver and has a more extensive effect on lowering serum bile acid.

R285.5

國(guó)家自然科學(xué)基金青年基金項(xiàng)目（81803874）