基于内源性大麻素系统探讨皂术茵陈方抗非酒精性脂肪性肝炎的作用机制

Non-alcoholic steatohepatitis (NASH), being a research focus recently, is one of important factors for development of liver cirrhosis, liver cancer and other chronic liver diseases. Results from previous studies have been proved that Zaozhuyinchen Decoction has the pharmacological effects of anti-NASH and the effect is closely related to the regulation of endogenous cannabinoids system(eCBs). On the basis of this, we put forward the scientific hypothesis that malfunction of eCBs is one of the important factors in the pathogenesis of NASH, the regulation of signal pathway of‘ERK/cAMP-CREB-BDNF’ maybe the key role of Zaozhuyinchen Decoction in treatment of NASH. In order to clarify the mechanism of Zaozhuyinchen Decoction for regulating on eCBs, NASH rat model induced by HFD, the in vitro and in vivo model of HEK-293-rFAAH,HEK-293-MGL cell lines will be applid in the following experiments. In the animal model, there are 40 rats will be divided into 4 groups: normal group, model group, Zaozhuyinchen Decoction group and Polyene Phosphatidylcholine Capsules group. Model group, Zaozhuyinchen Decoction group and Polyene Phosphatidylcholine Capsules group will be fed by HFD for 16 weeks and given corresponding drugs via intragastric administration for 8 weeks, respectively. HepG2 cell also will be individed into 3 groups, control group, model group and Zaozhuyinchen Decoction group. Both model group and Zaozhuyinchen Decoction group will be induced by FFA for 24 hr, in the mean time, Zaozhuyinchen Decoction group will be incubated with rat drug serumn.We will use lipidomics and molecular biology technology to discuss the disorder effect of the endogenous cannabinoid system on the NASH and to elucidate the mechanism of Zaozhuyinchen Decoction for NASH.

非酒精性脂肪性肝炎（NASH）作为肝纤维化、肝硬化、肝癌等慢性肝病发生、发展的重要因素，截断其发展已成为慢性肝病防治研究的焦点。皂术茵陈方以“清热利湿、健脾活血”功效，契合NASH“脾虚湿盛、热瘀互结”病机特点，具有显著的临床疗效。基于内源性大麻素系统参与NASH的发病机制，而皂术茵陈方具有调控内源性大麻素的药理效应，提出“内源性大麻素系统功能失调作为NASH的发病机制，调控其介导的‘ERK/cAMP-CREB-BDNF’信号通路或是皂术茵陈方治疗NASH的重要作用途径”科学假说。课题拟运用高脂饮食诱导的NASH大鼠及HEK-293-rFAAH、HEK-293-MGL细胞株、FFA诱导HepG2细胞脂肪变性的体内外模型，采用脂质组学、分子生物学等技术，阐明皂术茵陈方调控内源性大麻素系统的作用机制，剖析其对“ERK/cAMP-CREB-BDNF”信号通路的作用靶点。

研究背景：非酒精性脂肪性肝炎(NASH)作为肝纤维化、肝硬化、肝癌等慢性肝病发生、发展的重要因素，截断其发展已成为慢性肝病防治研究的焦点。皂术茵陈方以“清热利湿、健脾 活血”功效，契合NASH“脾虚湿盛、热瘀互结”病机特点，具有显著的临床疗效。基于内源性大麻素系统参与NASH的发病机制，而皂术茵陈方具有调控内源性大麻素的药理效应，提出“内源性大麻素系统功能失调作为NASH的发病机制，调控其介导的‘ERK/cAMP-CREB- BDNF’信号通路或是皂术茵陈方治疗NASH的重要作用途径”科学假说。.研究内容：运用高脂饮食诱导的NASH大鼠及HEK-293-rFAAH、HEK-293-MGL细胞株、FFA诱导HepG2细胞脂肪变性的体内外模型，采用脂质组学、分子生物学等技术，阐明皂术茵陈方调控内源性大麻素系统的作用机制，剖析其对“ERK/cAMP-CREB-BDNF”信号通路的作用靶点。.研究结果和意义：1、采用高脂饮食诱导的大鼠脂肪肝模型，证实皂术茵陈方具有显著治疗脂肪肝的药理效应，且具有降低内源性大麻素系统2-AG 、CB1、CB2含量、升高FAAH、MAGL表达的作用特点。2、采用HEK-293-rFAAH、HEK-293-MGL细胞株，进一步证实皂术茵陈方对FAAH 酶/MAGL酶具有直接的作用特点。3、采用高脂饮食诱导的大鼠脂肪肝模型及FFA诱导HepG2细胞脂肪变性体外模型，证实皂术茵陈方具有提高RAS、MEK、p-MEK、RSK、 ERK1/2、p-ERK1/2、PKA、p-CREB、BDNF蛋白表达的作用特点，说明调控‘ERK/cAMP-CREB- BDNF’信号通路是皂术茵陈方治疗NASH的重要作用途径。