背景:在全球范围内,非酒精性脂肪性肝病的发病率和患者数量不断增加,这已经成为人类健康的最大威胁之一。然而,到目前为止,仍然没有有效的治疗和药物。全叶SilphiumL.是一种原产于北美的多年生草本植物,用于改善体质和治疗印度部落传统草药中的肝脏和脾脏相关疾病。这种草本植物富含绿原酸,具有降低血脂的功能,减肥和保护肝脏。然而,这些化合物对非酒精性脂肪性肝病的影响尚不清楚.
目的:阐明绿原酸提取物(CY-10)及其类似物对非酒精性脂肪性肝病的治疗作用和机制,并确定活性化合物。
方法:建立游离脂肪酸诱导的HepG2细胞脂肪变性模型,评价CY-10促进脂质代谢的体外活性。Further,建立高脂饮食诱导的C57BL/6小鼠NAFLD模型,检测CY-10对小鼠各项生理生化指标的影响,并阐明提取物对调节脂质代谢的体内作用,抗炎和肝脏保护,并进行非目标脂质代谢组学分析肝脏中脂肪酸的差异代谢产物。随后,采用免疫印迹法和免疫组织化学方法对提取物的作用靶点进行分析,阐明其作用机制。最后,通过体外活性筛选阐明了CY-10中的活性化合物。
结果:结果表明,CY-10可显著减弱HepG2细胞的脂滴沉积。体内实验结果表明,CY-10能显著降低HFD诱导的小鼠体重和器官指数,改善生化指标,肝脏和血清中的氧化水平和炎症反应,从而保护肝脏组织。它可以促进肝脏中不饱和脂肪酸的代谢,减少饱和脂肪酸的生成。此外,阐明CY-10可通过调节AMPK/FXR/SREPB-1c/PPAR-γ信号通路促进脂质代谢平衡。最终,主要活性化合物被证明是隐绿原酸,对细胞内脂肪酸的代谢有很强的促进作用。令人印象深刻的是,在体内和体外,CY-10和隐绿原酸的活性均强于辛伐他汀。
结论:第一次,阐明了含药石中富含绿原酸及其类似物的提取物对非酒精性脂肪性肝病有较好的治疗作用。证实了隐绿原酸是主要活性化合物,具有良好的医药潜力。
BACKGROUND: Globally, the incidence rate and number of patients with nonalcoholic fatty liver disease are increasing, which has become one of the greatest threats to human health. However, there is still no effective therapy and medicine so far. Silphium perfoliatum L. is a perennial herb native to North America, which is used to improve physical fitness and treat liver and spleen related diseases in the traditional medicinal herbs of Indian tribes. This herb is rich in chlorogenic acids, which have the functions of reducing blood lipids, losing weight and protecting liver. However, the effect of these compounds on nonalcoholic fatty liver disease remains unclear.
OBJECTIVE: Clarify the therapeutic effects and mechanism of the extract (CY-10) rich in chlorogenic acid and its analogues from Silphium perfoliatum L. on non-alcoholic fatty liver disease, and to determine the active compounds.
METHODS: A free fatty acid-induced steatosis model of HepG2 cells was established to evaluate the in vitro activity of CY-10 in promoting lipid metabolism. Further, a high-fat diet-induced NAFLD model in C57BL/6 mice was established to detect the effects of CY-10 on various physiological and biochemical indexes in mice, and to elucidate the in vivo effects of the extract on regulating lipid metabolism, anti-inflammation and hepatoprotection, and nontarget lipid metabolomics was performed to analyze differential metabolites of fatty acids in the liver. Subsequently, western blotting and immunohistochemistry were used to analyze the target of the extract and elucidate its mechanism of action. Finally, the active compounds in CY-10 were elucidated through in vitro activity screening.
RESULTS: The results indicated that CY-10 significantly attenuated lipid droplet deposition in HepG2 cells. The results of in vivo experiments showed that CY-10 significantly reduce HFD-induced mouse body weight and organ index, improve biochemical indexes, oxidation levels and inflammatory responses in the liver and serum, thereby protecting the liver tissue. It can promote the metabolism of unsaturated fatty acids in the liver and reduce the generation of saturated fatty acids. Furthermore, it is clarified that CY-10 can promote lipid metabolism balance by regulating AMPK/FXR/SREPB-1c/PPAR-γ signal pathway. Ultimately, the main active compound was proved to be cryptochlorogenic acid, which has a strong promoting effect on the metabolism of fatty acids in cells. Impressively, the activities of CY-10 and cryptochlorogenic acid were stronger than simvastatin in vitro and in vivo.
CONCLUSIONS: For the first time, it is clarified that the extract rich in chlorogenic acids and its analogues in Silphium perfoliatum L. have good therapeutic effects on non-alcoholic fatty liver disease. It is confirmed that cryptochlorogenic acid is the main active compound and has good potential for medicine.