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目的观察经脂多糖(LPS)诱导后连续给予何首乌醇提液(AEP)7 d大鼠的肝损伤程度及胆汁淤积相关指标的变化。方法将雄性SD大鼠随机分为5组:对照组、LPS组、LPS+对乙酰氨基酚(APAP)组、AEP组、LPS+AEP组,LPS、LPS+APAP、LPS+AEP组按大鼠体质量分别尾iv给予4 mg/kg LPS,对照组与AEP组给予等体积生理盐水;2 h后LPS+APAP组ig给予625 mg/kg APAP,AEP组与LPS+AEP组ig给予12 g/kg,每天1次,连续给药7 d,建立特异质肝损伤炎症模型。观察体质量变化,并分别于造模后2 h、14 h、5 d、8 d检测血清生化中肝功能相关指标的变化,同时收集胆汁,计算胆汁流速与密度,并检测胆汁中主要成分变化。进行肝脏系数及组织病理学检查,并对胆汁淤积相关蛋白胆盐输出泵转运蛋白(BSEP)、多药耐药蛋白2(MRP2)及多药耐药蛋白3(MRP3)进行Real-Time PCR检测。结果经过LPS诱导2 d后,LPS+AEP组与对照组、AEP组比较体质量明显下降,5、8 d肝脏系数显著增加(P<0.05);血清生化指标分析结果显示,与对照组比较,LPS+AEP组丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)水平无明显变化;造模后8 d的总胆红素(TBIL)明显降低(P<0.05),ALP明显升高(P<0.05);可观察到胆汁密度和胆汁流速明显降低(P<0.05),对胆汁成分分析显示,与对照组比较,LPS+AEP组总胆固醇(TCHO)显著升高、TBIL显著降低(P<0.05);病理结果显示,AEP组出现轻微肝细胞变性,而LPS+AEP组可见严重局灶坏死;转录水平结果发现,LPS诱导后可使得BSEP、MRP2在14 h时出现短期抑制(P<0.05),单独给予AEP可使BSEP、MRP2和MRP3的表达水平短时显著升高(P<0.05、0.01),而LPS+AEP给药第8 d对大鼠肝脏BSEP、MRP2无显著性影响,可使MRP3转录水平升高(P<0.05)。结论经LPS诱导的AEP可明显损伤大鼠肝细胞并干扰胆汁分泌功能,引起胆汁成分相关生化指标的改变,对BSEP与MRP2水平无显著影响,MRP3出现代偿性升高,提示确实存在胆汁淤积症状,但可能是存在其他机制。
Objective To observe the changes of hepatic injury and related indicators of cholestasis after continuous administration of alcohol extract of Polygonum multiflorum (AEP) induced by lipopolysaccharide (LPS) for 7 days. Methods Male SD rats were randomly divided into 5 groups: control group, LPS group, LPS + acetaminophen (APAP) group, AEP group, LPS + AEP group, LPS, LPS + APAP, LPS + The rats in the LPS + APAP group were given 625 mg / kg APAP after 2 h, and those in the AEP group and LPS + AEP group were given 12 g / kg , Once a day for 7 consecutive days to establish a model of liver injury with specific liver inflammation. The change of body weight was observed and the changes of liver function related indexes in serum biochemistry were detected at 2 h, 14 h, 5 d, 8 d after modeling respectively. At the same time, bile was collected, the bile flow rate and density were calculated, and the changes of the main components in bile . The liver coefficient and histopathological examination were performed. Real-time PCR was used to detect the expression of bile duct-associated protein bile salt export pump transporter (BSEP), multidrug resistance protein 2 (MRP2) and multidrug resistance protein 3 (MRP3) . Results After 2 days of LPS induction, the body weight of LPS + AEP group decreased significantly compared with that of AEP group and liver index increased 5 and 8 days (P <0.05). The results of serum biochemical analysis showed that compared with control group, There was no significant change in ALT and AST levels in LPS + AEP group. The total bilirubin (TBIL) was significantly decreased on the 8th day after modeling (P <0.05) ALP significantly increased (P <0.05); bile density and bile flow rate were significantly decreased (P <0.05). Analysis of bile composition showed that total cholesterol (TCHO) increased significantly in LPS + AEP group compared with control group TBEP significantly decreased (P <0.05) .Pathological examination showed slight hepatocellular degeneration in AEP group and severe focal necrosis in LPS + AEP group. Transcriptional levels of BSEP and MRP2 were found to be significant at 14 h after LPS induction (P <0.05). The expression of BSEP, MRP2 and MRP3 were significantly increased in short time (P <0.05, 0.01) by AEP alone, while the expression of BSEP, MRP2 No significant effect, can make MRP3 transcription level increased (P <0.05). Conclusion AEP induced by LPS can obviously damage the rat hepatocytes and interfere with the bile secretion function, cause the change of biochemical indexes related to bile components, have no significant effect on the levels of BSEP and MRP2, and MRP3 appear compensatory increase, suggesting the existence of cholestasis Symptoms, but there may be other mechanisms.