Department of Pharmacognosy, China Pharmaceutical University, Nanjing 201009, China.
J Ethnopharmacol. 2012 Mar 27;140(2):222-9. doi: 10.1016/j.jep.2011.12.047. Epub 2012 Jan 16.
Danning tablet, as a composite prescription of traditional Chinese medicine, has been used clinically to relieve liver and gallbladder diseases in China. However, the mechanisms involved are still unclear.
The present investigation was designed to assess the effects and possible mechanisms of Danning tablet on α-naphthylisothiocyanate (ANIT)-induced liver injury with cholestasis.
Danning tablet (3, 1.5 or 0.75g/kg body weight/day) was intragastrically (i.g.) given to experimental rats for seven days before they were treated with ANIT (60mg/kg daily via i.g.) which caused liver injury. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), γ-glutamyltranspeptidase (γ-GTP), total bilirubin (T-Bil), direct bilirubin (D-Bil), total bile acid (TBA) and bile flow were measured to evaluate the protective effect of Danning tablet at 48h after ANIT treatment. Furthermore, protective mechanisms of Danning tablet against ANIT-induced liver injury were elucidated by assays of liver enzyme activities and component contents including myeloperoxidase (MPO), superoxide dismutase (SOD), glutathione peroxidase (Gpx), catalase (CAT) and glutathione S-transferase (GST), as well as liver lipid peroxide (LPO) and glutathione (GSH). The biochemical observations were supplemented by histopathological examination. Phytochemical analysis of Danning tablet was performed by UPLC-MASS.
Obtained results demonstrated that high dose (3g/kg) of Danning tablet significantly prevented ANIT-induced changes in bile flow (P<0.01), and serum levels of ALT, AST, ALP, γ-GTP, T-Bil, D-Bil (P<0.01) and TBA (P<0.05). In addition, ANIT-induced increases in hepatic MPO, GST activities and GSH, LPO contents were significantly (P<0.01) reduced, while SOD, Gpx, CAT activities in the liver tissue which were suppressed by ANIT were significantly (P<0.01) elevated in the groups pretreated with Danning tablet at the dose of 3g/kg B.W. Histopathology of the liver tissue showed that pathological injuries were relieved after Danning tablet (3g/kg) pretreatment. The results also showed that medium dose (1.5g/kg) of Danning tablet exhibited partially protective effect on ANIT-induced liver injury with cholestasis by reversing part of biochemical parameters and histopathological changes. Low dose (0.75g/kg) of Danning tablet did not show any protective effect on ANIT-induced liver injury with cholestasis. Phytochemical analyses revealed the presence of anthraquinones, flavonoids and stilbene in the Danning tablet.
These findings indicate that Danning tablet exerts a dose-dependently protective effect on ANIT-induced liver injury with cholestasis in rats, and the possible mechanism of this activity is likely due to its attenuation of oxidative stress in the liver tissue and neutrophil infiltration.
丹宁片作为一种中药复方，已在临床上用于缓解中国肝胆疾病。然而，其涉及的机制仍不清楚。
本研究旨在评估丹宁片对α-萘异硫氰酸酯（ANIT）诱导的胆汁淤积性肝损伤的作用及可能机制。
丹宁片（3、1.5 或 0.75g/kg 体重/天）连续 7 天灌胃给药，然后用 ANIT（60mg/kg 每日灌胃）处理实验大鼠，ANIT 可引起肝损伤。在 ANIT 处理后 48 小时，测定血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、碱性磷酸酶（ALP）、γ-谷氨酰转肽酶（γ-GTP）、总胆红素（T-Bil）、直接胆红素（D-Bil）、总胆汁酸（TBA）和胆汁流量，以评估丹宁片的保护作用。此外，通过测定肝酶活性和丙二醛（LPO）、谷胱甘肽（GSH）含量以及髓过氧化物酶（MPO）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（Gpx）、过氧化氢酶（CAT）和谷胱甘肽 S-转移酶（GST）的含量，阐明丹宁片对 ANIT 诱导的肝损伤的保护机制。肝脂质过氧化（LPO）。生化观察结果辅以组织病理学检查。采用 UPLC-MASS 对丹宁片进行植物化学分析。
结果表明，高剂量（3g/kg）丹宁片能显著预防 ANIT 诱导的胆汁流量变化（P<0.01），并显著降低血清 ALT、AST、ALP、γ-GTP、T-Bil、D-Bil（P<0.01）和 TBA（P<0.05）水平。此外，ANIT 诱导的肝 MPO、GST 活性和 GSH、LPO 含量增加明显（P<0.01），而 SOD、Gpx、CAT 活性则明显升高（P<0.01）。肝组织中被 ANIT 抑制的 CAT 活性。丹宁片（3g/kg）预处理组的组织病理学检查显示，肝组织病理损伤得到缓解。结果还表明，中剂量（1.5g/kg）丹宁片通过逆转部分生化参数和组织病理学变化，对 ANIT 诱导的胆汁淤积性肝损伤具有部分保护作用。低剂量（0.75g/kg）丹宁片对 ANIT 诱导的胆汁淤积性肝损伤无保护作用。植物化学分析表明，丹宁片中含有蒽醌、黄酮和芪类化合物。
这些发现表明，丹宁片对 ANIT 诱导的大鼠胆汁淤积性肝损伤具有剂量依赖性的保护作用，其活性的可能机制可能是通过减轻肝组织氧化应激和中性粒细胞浸润。
