化痰清利法对非酒精性脂肪性肝炎大鼠JNK1网络信号通路的调控机制研究

Bei Jing University of Chinese Medicine’s innovation team project(No.2011-CXTD-24) has found that Erchen Tang and Yinchenhao Tang can regulate the expression of TNF-α and IRS-1Ser307 of non-alcoholic steatohepatitis (NASH) model, and Erchen Tang can regulate the expression of IRS-1Ser307. TNF-α、IRS-1、IRS-1Ser307 are important correlation factors of JNK1 net signaling pathway. We use NASH model in rats established by feeding high fat diet and hepatocyte model in rats vitro to study the molecular mechanism of JNK1 net signaling pathway on NASH by resolving phlegm and clearing therapy. We take a lead research on molecular mechanism of JNK1 pathway in rats with NASH，and make a detail research on the expression of JNK1, p-IRs-1Ser307, IRs-1, p-PKBSer473, PKB, TNF-α, AP-1 in JNK1 net signaling pathway，which could confirm targets of NASH by resolving phlegm and clearing therapy, uncovering the the mechanism of regulating liver IR and improving liver inflammation by JNK1 net signaling pathway from the perspective of molecular biology. Meanwhile, the difference in regulating JNK1 net pathway would be explicated by comparing their difference between HuatanQingli Decoctionon, Rosiglitazone and Essentiale which prevent and treat NASH. The study will make great contribution to clinical use of resolving phlegm and clearing therapy.

北京中医药大学创新团队项目（No.2011-CXTD-24）研究发现：化痰法的代表方二陈汤和清利法的代表方茵陈蒿汤可调控NASH大鼠JNK1网络信号通路中的相关因子：TNF-α、IRS-1、IRS-1Ser307的表达。本课题分别采用高脂饮食和软脂酸建立NASH大鼠模型和体外大鼠肝细胞模型，研究化痰清利法对JNK1网络信号通路的调控机制，重点研究JNK1网络信号通路中肝脏JNK1、p-IRs-1Ser307、IRs-1、肝脏p-PKBSer473及PKB、TNF-α、AP-1等多种蛋白的表达，明确化痰清利法防治NASH的作用靶点，从分子生物学角度揭示化痰清利法通过JNK1网络信号通路调控肝脏IR，改善肝组织炎症的机制。同时，比较化痰清利法的代表方与罗格列酮、易善复防治NASH的疗效差异，明确其对JNK1网络信号通路的作用异同，阐明作用靶点的异同，为化痰清利法防治NASH提供科学依据。

近年来，中国的非酒精性脂肪性肝病的发病率呈不断上升趋势。非酒精性脂肪性肝炎，是病情发展及进一步演变的重要阶段。中医药在改善肝功能、调节脂质代谢上具有比较明显的优势和确切的疗效。运用化痰清利法的代表方茵陈二陈汤（茵陈蒿汤和二陈汤合方），对NASH动物模型和细胞模型进行干预，并以罗格列酮组、多烯磷脂酰胆碱组作为对照。结果表明：①化痰清利法可通过有效改善NASH模型大鼠肝功能及纠正肝组织细胞氧化-抗氧化平衡紊乱，降低模型大鼠肝组织内TNF水平，缓解模型大鼠肝组织内脂肪沉积、肝细胞脂肪变性及抑制肝组织炎症，进而缓解大鼠肝组织损伤，发挥干预治疗作用。②化痰清利法可下调NASH模型IRS-1ser307蛋白及IRS-1ser307／IRS-1水平，上调PKBser473蛋白及PKBser473／PKB水平，通过下调IRS-1丝氨酸磷酸化水平，进而上调其下游PKB活化水平，发挥对模型大鼠糖脂代谢调控及提高模型大鼠胰岛素敏感性的干预治疗作用。③化痰清利法可有效降低NASH模型JNK1 mRNA水平及JNK1蛋白水平，抑制NASH模型AP-1 mRNA水平及AP-1蛋白水平升高，通过下调JNK1 mRNA及JNK1蛋白表达水平，并作用于下游信号IRS-1、PKB，影响下游信号活化水平，进而缓解胰岛素抵抗，纠正糖脂代谢紊乱；通过下调JNK1 mRNA及JNK1表达水平，进而降低AP-1 mRNA水平及AP-1蛋白，进而发挥降低肝组织TNF水平、抑制肝组织炎症损伤干预作用。