Abstract：
BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease with therapeutic options on the horizon. Picrorhiza kurroa, enriched with iridoid glycosides like picroside I and picroside II is known for its hepatoprotective activity and anti-inflammatory properties. Androsin, the other phytochemical present in P. kurroa has been shown to have anti-inflammatory and anti-asthmatic properties. However, its role in NAFLD is yet to be investigated.
OBJECTIVE: This study aims to identify the potent hepatoprotective agent from P. kurroa that can attenuate NAFLD in HFrD-fed ApoE-/- mice, and elucidate the underlying mechanisms governing its effects.
METHODS: Classical purification methods were used to isolate seven compounds, including picroside I, picroside II and androsin from the roots of P. kurroa. NAFLD-induced ApoE-/- mice were administered orally with either picroside I, picroside II, or androsin for 7 weeks. Animals were scanned non-invasively by ultrasonography at 1st and 14th week. Gross histomorphometry was examined by HE and Sirius red staining. mRNA transcript and protein profile associated with autophagy, lipogenesis, inflammation, and fibrosis was done through RT-PCR and Western blot analysis.
RESULTS: In-vitro and in-vivo studies revealed that among the seven evaluated compounds, androsin shows the most potent in-vitro activity. Oral dosing of androsin (10 mg/kg) protected the liver against HFrD-induced NAFLD in ApoE-/- mice model. Biochemical analysis revealed a reduction in ALT and AST enzymes and a significant reduction in cholesterol levels. Hepatocyte ballooning, hepatic lipid deposition, inflammation, and fibrosis were reduced. Androsin treatment significantly reduced fibrosis (α-SMA, collagens, TGF-β) and inflammation (ILs, TNF-α, NFκB) in ApoE-/- mice. Mechanistically, androsin activated AMPKα and down-regulated the expression of SREBP-1c, resulting in ameliorating hepatic lipogenesis.
CONCLUSIONS: Our results support autophagy as one of the therapeutic strategies to reduce steatosis and hepatic damage. We found that androsin treatment significantly ameliorated hepatic steatosis, serum lipid levels, and hepatic injury in ApoE-/- induced by HFrD. Androsin administration mitigated lipogenesis by inhibiting SREBP1c/FASN pathway and activating autophagy through AMPKα/PI3K/Beclin1/LC3 pathway.
OBJECTIVE: This study aims to identify the potent hepatoprotective agent from P. kurroa that can attenuate NAFLD in HFrD-fed ApoE-/- mice, and elucidate the underlying mechanisms governing its effects.
METHODS: Classical purification methods were used to isolate seven compounds, including picroside I, picroside II and androsin from the roots of P. kurroa. NAFLD-induced ApoE-/- mice were administered orally with either picroside I, picroside II, or androsin for 7 weeks. Animals were scanned non-invasively by ultrasonography at 1st and 14th week. Gross histomorphometry was examined by HE and Sirius red staining. mRNA transcript and protein profile associated with autophagy, lipogenesis, inflammation, and fibrosis was done through RT-PCR and Western blot analysis.
RESULTS: In-vitro and in-vivo studies revealed that among the seven evaluated compounds, androsin shows the most potent in-vitro activity. Oral dosing of androsin (10 mg/kg) protected the liver against HFrD-induced NAFLD in ApoE-/- mice model. Biochemical analysis revealed a reduction in ALT and AST enzymes and a significant reduction in cholesterol levels. Hepatocyte ballooning, hepatic lipid deposition, inflammation, and fibrosis were reduced. Androsin treatment significantly reduced fibrosis (α-SMA, collagens, TGF-β) and inflammation (ILs, TNF-α, NFκB) in ApoE-/- mice. Mechanistically, androsin activated AMPKα and down-regulated the expression of SREBP-1c, resulting in ameliorating hepatic lipogenesis.
CONCLUSIONS: Our results support autophagy as one of the therapeutic strategies to reduce steatosis and hepatic damage. We found that androsin treatment significantly ameliorated hepatic steatosis, serum lipid levels, and hepatic injury in ApoE-/- induced by HFrD. Androsin administration mitigated lipogenesis by inhibiting SREBP1c/FASN pathway and activating autophagy through AMPKα/PI3K/Beclin1/LC3 pathway.
摘要：
背景：非酒精性脂肪性肝病（NAFLD）是一种慢性肝病，具有治疗选择。Picrorhizakurroa,富含环烯醚萜类糖苷如胡黄连苷I和胡黄连苷II以其保肝活性和抗炎特性而闻名。Androsin,另一种植物化学物质存在于P.kurroa已被证明具有抗炎和抗哮喘的特性。然而,其在NAFLD中的作用尚待研究。
目的：本研究旨在确定来自P.kurroa的有效的肝保护剂，可以减弱HFrD喂养的ApoE-/-小鼠的NAFLD，并阐明其影响的潜在机制。
方法：经典纯化方法用于分离七个化合物，包括胡黄连苷I,来自P.kurroa的根的picrosideII和雄香。NAFLD诱导的ApoE-/-小鼠口服给予胡黄连苷I，黄连苷Ⅱ,或雄激素7周。在第1周和第14周通过超声非侵入性扫描动物。通过HE和天狼星红染色检查大体组织形态计量学。与自噬相关的mRNA转录和蛋白质谱，脂肪生成,炎症,通过RT-PCR和Westernblot分析进行纤维化。
结果：体外和体内研究表明，在七个被评估的化合物中，雄激素显示出最有效的体外活性。在ApoE-/-小鼠模型中,口服给药雄素(10mg/kg)保护肝脏免受HFrD诱导的NAFLD。生化分析显示ALT和AST酶的降低和胆固醇水平的显著降低。肝细胞膨胀，肝脂质沉积，炎症,纤维化减少。雄激素治疗显着减少纤维化（α-SMA，胶原蛋白，TGF-β）和炎症（IL，TNF-α,NFκB）在ApoE-/-小鼠中。机械上,雄激素激活AMPKα并下调SREBP-1c的表达，导致改善肝脏脂肪生成。
结论：我们的结果支持自噬作为减少脂肪变性和肝损伤的治疗策略之一。我们发现，雄激素治疗显著改善肝脂肪变性，血脂水平，和HFrD诱导的ApoE-/-肝损伤。雄激素通过抑制SREBP1c/FASN途径和通过AMPKα/PI3K/Beclin1/LC3途径激活自噬来减轻脂肪生成。
目的：本研究旨在确定来自P.kurroa的有效的肝保护剂，可以减弱HFrD喂养的ApoE-/-小鼠的NAFLD，并阐明其影响的潜在机制。
方法：经典纯化方法用于分离七个化合物，包括胡黄连苷I,来自P.kurroa的根的picrosideII和雄香。NAFLD诱导的ApoE-/-小鼠口服给予胡黄连苷I，黄连苷Ⅱ,或雄激素7周。在第1周和第14周通过超声非侵入性扫描动物。通过HE和天狼星红染色检查大体组织形态计量学。与自噬相关的mRNA转录和蛋白质谱，脂肪生成,炎症,通过RT-PCR和Westernblot分析进行纤维化。
结果：体外和体内研究表明，在七个被评估的化合物中，雄激素显示出最有效的体外活性。在ApoE-/-小鼠模型中,口服给药雄素(10mg/kg)保护肝脏免受HFrD诱导的NAFLD。生化分析显示ALT和AST酶的降低和胆固醇水平的显著降低。肝细胞膨胀，肝脂质沉积，炎症,纤维化减少。雄激素治疗显着减少纤维化（α-SMA，胶原蛋白，TGF-β）和炎症（IL，TNF-α,NFκB）在ApoE-/-小鼠中。机械上,雄激素激活AMPKα并下调SREBP-1c的表达，导致改善肝脏脂肪生成。
结论：我们的结果支持自噬作为减少脂肪变性和肝损伤的治疗策略之一。我们发现，雄激素治疗显著改善肝脂肪变性，血脂水平，和HFrD诱导的ApoE-/-肝损伤。雄激素通过抑制SREBP1c/FASN途径和通过AMPKα/PI3K/Beclin1/LC3途径激活自噬来减轻脂肪生成。
