Pure total flavonoids from citrus improve nonalcoholic steatohepatitis liver inflammatory responses by regulating the CCL2/CCR2-PI3K-Akt signal transduction pathway

Nonalcoholic steatohepatitis (NASH) is a critical stage in the prognosis of nonalcoholic fatty liver disease (NAFLD). Pure total flavonoids from circus (PTFC) play essential roles in the improvement of NASH symptoms, but the underlying regulatory mechanism remains elusive. Our previous high-throughput omics screening

A mice model of NASH was established by a high-fat diet, and PTFC was used as treatment. Hematoxylin-eosin and oil red O staining were used to observe the pathological changes in the liver tissue. Western blotting and real-time PCR were used to measure the mRNA and protein levels in the liver. The expression of proinflammatory cytokines in the peripheral blood and liver tissues was measured by liquid suspension array. An automatic biochemical method was used to examine serum transaminases and lipids levels, as well as liver lipids.

Compared with the mice in the high-fat diet group, mice in the HFD + PTFC group showed significantly improved liver histopathology, and levels of serum transaminase and lipids, liver lipids and serum proinflammatory cytokines. Moreover, the mRNA and protein expression and phosphorylation levels of key signaling molecules in the CCL2/CCR2-PI3K-Akt signal transduction pathway were obviously reduced by PTFC treatment. Conclusive remarks: PTFC can ameliorate NASH symptoms, and the mechanism may be related to regulating the CCL2/CCR2-PI3K-Akt signal transduction pathway to reduce the liver inflammatory response. 背景和目标: 非酒精性脂肪性肝炎 (NASH) 是非酒精性脂肪性肝病 (NAFLD) 的重要疾病阶段。胡柚皮黄酮 (PTFC) 在改善 NASH 各种症状中扮演着重要角色, 但是其具体的调控机制仍未研究清楚。我们前期的高通量研究显示, CCL2/CCR2-PI3K-Akt 信号通路是调控肝脏炎症反应的关键信号通路。PTFC可能通过调控 CCL2/CCR2-PI3K-Akt 信号通路来改善 NASH 肝脏炎症反应。. 方法: 通过高脂饮食建立NASH动物模型, 并用 PTFC 进行干预。运用HE染色和油红 O 染色来观察肝脏的组织病理学变化; 利用 Western blotting 技术 和 real time- PCR 技术来检测肝脏的基因和蛋白的表达; 采用液相悬浮芯片技术来检测肝脏和血清的促炎细胞因子的表达; 自动生化法检查肝脏和血清的转氨酶水平和脂质水平。. 结果: 与模型组相比, PTFC 干预组小鼠的肝脏病理组织学, 血清和肝脏转氨酶、脂质水平, 以及促炎细胞因子明显改善。此外, CCL2/CCR2-PI3K-Akt 信号通路中关键分子的基因和蛋白表达以及磷酸化水平较正常组均得到明显的改善。. 结论: PTFC 能改善 NASH 的各种临床症状, 其机制可能通过调控 CCL2/CCR2-PI3K-Akt 信号通路来减少肝脏的炎症反应。.