洛神花萃取物對Acetaminophen引發
肝損傷之保護作用

Acetaminophen(AAP)在社會中已成為普遍使用解熱鎮痛的成藥，不當的使用會經由氧化傷害導致肝受損。AAP代謝機制是經由體內的cytochrome P450代謝成NAPQI，促使細胞受損。洛神花則在先前的研究中證實具有強效的抗氧化作用，能清除體內過多的自由基、降低氧化傷害。因此本實驗利用洛神花多酚（HPE）及粗萃物（HSE）來預防AAP所誘導的肝受損。在BABL/c mice的動物實驗中‚連續兩週每日以餵管的方式給予HPE 100mg/kg、200mg/kg、300mg/kg及HSE 200mg/kg、400mg/kg、600mg/kg‚再以腹腔注射過量的AAP 1000mg/kg誘導肝受損，六小時後犧牲取肝臟做分析。在BABL/c normal liver cell的細胞實驗中，分別加入不同濃度的HPE及HSE (0.05、0.1、0.5、1 mg/ml)，再加入AAP 5mM以誘導細胞受損，並探討其蛋白表現分析及作用機制。實驗結果發現，在動物實驗中經HPE(100mg/kg、200mg/kg、300mg/kg)及HSE(200mg/kg、400mg/kg、600mg/kg)處理後，血清中肝功能指數有明顯減少，並降低肝臟脂質過氧化程度(TBARS)，分別減少63.2%,70.1%,83.1%及70.6%,65.1%,54.1% ;增加抗氧化性酵素或抗氧化分子，如過氧化酶(catalase)分別增加22%,24.4%,16.2%及45.7%,44.3%,55.8%；增加GSH分別為43.4%,25.2%,27.9%及41.5%,45.7%,54.2% ;在肝臟病理切片中，可減緩由APAP引起的肝受損程度。在動物實驗及細胞實驗中，進一步分析蛋白表現，經洛神HPE及HSE處理，可減少由APAP誘導的JNK、iNOS、Bax、Bid。在細胞實驗中當加入JNK抑制劑，也確實可抑制APAP所誘導的細胞受損。可知APAP是藉由活化JNK造成細胞受損，經由HPE及HSE處理後亦可透過抑制JNK的活化，進而抑制iNOS的活化，並抑制Bid、Bax的活化，減緩APAP所誘導的肝受損。由上述結果，我們推測洛神花多酚（HPE）及粗萃物（HSE）對於APAP所誘導的肝受損有保護效果，且此保護作用可能是經由JNK的抑制作用而導致下游蛋白改變而達成。

關鍵字

乙醯酚氨；洛神花

並列摘要

Acetaminophen (AAP) possessed analgesic and antipyretic properties; but it could induce hepatic necrosis in centrilobular cell. The major cause of hepatic damage was contributed by the reactive intermediate N-acetyl-p-benzoquinone imine (NAPQI). NAPQI accumulation was due to protein arylation and severe oxidative stress to cause liver injury. It was demonstrated that Hibisus Sabdariffa (HS) potentiated antioxidative effect in previous studies. In this study, we investigated HS extracts against AAP induced liver injury in BABL/c mice. In vivo, BABL/c mice were fed orally with HS polyphenols extract (HPE) (100, 200, 300 mg/kg) or HS aqueous extract (HSE) (200, 400, 600 mg/kg) for two weeks; and then 1000mg/kg of AAP was injected i.p. At 6h after APAP injection, mice were decapitated. In vitro, BABL/c normal liver (BNL) cell was pretreated with AAP 5mM or HPE/HSE (0.05, 0.1, 0.5, 1 mg/kg) for 48 hrs. Base on that, we investigated the possible mechanism. The results showed that after AAP administration, pretreatment of HPE (100, 200, 300 mg/kg) or HSE (200, 400, 600 mg/kg) decreased the levels of lipid peroxidation to 63.2%, 70.1%, 83.1% (HPE) or 70.6%, 65.1%, 54.1%(HSE) ；catalase increased to 22%, 24.4%, 16.2% (HPE) or 45.7%, 44.3%, 55.8%(HSE), glutathione increased to 43.4%, 25.2%, 27.9%(HPE) or 41.5%, 45.7%, 54.2%(HSE). In histopathological evaluation of liver, HSE or HPE can decrease AAP induced liver injury. Both in vivo and in vitro, HSE or HPE can decreased the expressions of JNK, iNOS, Bax, Bid. Futhermore in vitro assay, we also found that AAP induced cell injury could be decreased by JNK inhibitor as well as HSE/HPE. In conclusion, we have demonstrated that HPE or HSE can protect liver from AAP induced injury. The protecting mechanism could involve inhibition of (1) JNK1/2 activation coupled with inhibition of Bax/Bid activation; (2) induction of oxidative stress.

並列關鍵字

Acetaminophen ； Hibiscus sabdariffa

參考文獻

1. Xu, L.M. and Y.Y. Hu, [Studies on treatment of fatty liver with traditional Chinese medicine]. Zhong Xi Yi Jie He Xue Bao, 2003. 1(2): p. 138-41.

2. Zhang, J. and L.M. Xu, [Advances of mechanism research on treatment of experimental liver fibrosis with traditional Chinese medicine]. Zhong Xi Yi Jie He Xue Bao, 2003. 1(2): p. 142-5.

3. Kon, K., et al., Mitochondrial permeability transition in acetaminophen-induced necrosis and apoptosis of cultured mouse hepatocytes. Hepatology, 2004. 40(5): p. 1170-9.

4. Ben-Av, P., et al., Induction of vascular endothelial growth factor expression in synovial fibroblasts by prostaglandin E and interleukin-1: a potential mechanism for inflammatory angiogenesis. FEBS Lett, 1995. 372(1): p. 83-7.

5. Ouellet, M. and M.D. Percival, Mechanism of acetaminophen inhibition of cyclooxygenase isoforms. Arch Biochem Biophys, 2001. 387(2): p. 273-80.

國際替代計量

洛神花萃取物對Acetaminophen引發肝損傷之保護作用

全文下載

主題瀏覽