补肾升脾方调节ob/ob小鼠脂肪组织中脂肪因子表达的研究

Obesity is a kind of chronic low-grade inflammation condition which is closely related to adipocytokines disorder, arteriosclerosis and insulin resistance. Because of the fewer side effects, the traditional Chinese medicine is regarded as the first clinical choice of treatment for obesity. The previous treatments are mainly in strengthening spleen and benefiting qi, drying dampness and eliminating phlegm. In recent years, there are some progress in the aspect of theory, while the effect is not good. According to the previous studies, we introduced the hypothesis of depressed essence and qi that qi transformation would be depressed due to renal deficiency and qi falling would happen due to splenic asthenia, which might solve both the contradiction between phlegmatic hygrosis due to splenic asthenia and redundantly ingesting and surplus energy and formation mechanism of qi deficiency, phlegmatic hygrosis, qi depression and blood stasis. In the course of study, the ob/ob mouse models would be intervened by the decoction for invigorating kidney and lifting spleen. After that, QRT-PCR technology would be used to test the mRNA expression of adiponectin and MAPK in adipose tissue. Additionally, the concentration of adipose tissue and the blood plasma would be detected by Western blotting and enzyme-linked immunosorbent assay respectively.The distinction among invigorating kidney ,lifting spleen and invigorating spleen to remove phlem would be discovered by observing the decoction' influence on adipocytokines in gene, protein and endocrine aspects. The interaction among invigorating kidney and lifting spleen would be analyzed by Factor Analysis Approach. We hope that the hypothesis of depressed essence and qi would be proved, the theories of mechanism of obesity would be developed, and the guidance for clinical treatment and drug development would be provided.

肥胖是一种慢性低度炎症状态，该炎症状态及脂肪因子紊乱与动脉硬化、胰岛素抵抗等密切相关，中医药治疗因副作用少而为临床首选。传统治法以健脾益气、燥湿化痰为主，近年来虽有一定进展，但效果并不理想。我们根据既往结果，提出"肾虚气化减退、脾虚清阳不升"的"精气郁滞"病机假说。既理顺了脾虚痰湿与摄食亢进、能量过剩的矛盾，也解释了气虚、痰湿、气滞、血瘀的形成机制。 本项目拟采用ob/ob小鼠，以补益肾气、健脾升提的补肾升脾方等进行干预，实时荧光定量PCR检测脂肪组织中脂联素等代谢因子、MAPK等炎症因子mRNA表达；蛋白免疫印迹检测脂肪组织、酶联免疫检测血清中浓度（含量），从基因、蛋白、内分泌个方面观察补肾升脾方对脂肪因子的影响；比较补肾、升脾与健脾化痰的差异，利用两因素析因分析法分析补肾与升脾的交互作用，验证肾虚不化、脾虚不升的"精气郁滞"病机假说，发展肥胖病机理论，为临床治疗和药物开发提供指导。

肥胖是一种慢性低度炎症状态，该炎症状态及脂肪因子紊乱与动脉硬化、胰岛素抵抗等密切相关，中医药治疗因副作用少而为临床首选。我们根据既往结果，提出“肾虚气化减退、脾虚清阳不升”的“精气郁滞”病机假说。既理顺了脾虚痰湿与摄食亢进、能量过剩的矛盾，也解释了气虚、痰湿、气滞、血瘀的形成机制。因此，本项目以期阐明肥胖的根本病理机制是肾不气化、脾不升清、精微郁滞。 .本项目拟采用ob/ob小鼠，以补益肾气、健脾升提的补肾升脾方等进行干预，利用实时荧光定量PCR检测脂肪组织中脂联素、抵抗素等代谢相关因子，TNF-α等炎症相关因子的mRNA表达；利用酶联免疫法测定脂肪组织和血清中的脂联素、抵抗素和胰岛素的浓度（含量），从基因、内分泌方面观察补肾升脾方对脂肪因子的影响；比较补肾、升脾与健脾化痰的差异，验证肾虚不化、脾虚不升的“精气郁滞”病机假说。.研究发现①ob/ob小鼠体重下降，脂肪因子Adiponectin mRNA表达升高，Resistin和TNF-a mRNA的表达下降。脂肪和血清中的胰岛素下降，脂联素和抵抗素上升。②补肾升脾组小鼠内脏和皮下脂肪中脂肪因子Adiponectin mRNA的表达明显高于其他中药治疗组；内脏和皮下脂肪组织Resistin mRNA及TNF-αmRNA的表达明显低于其他中药治疗组； 血清胰岛素和皮下脂联素 内脏抵抗素 以补肾升脾组 最佳，且皮下脂联素提示补肾升脾药物之间存在交互作用。.结论：加味六君子汤具有调节ob/ob小鼠脂肪因子表达的作用，补肾、升脾药物存在复杂的交互作用，以补肾升脾效果最佳；分析补肾升脾药物的交互作用，优化配伍组合，对于提高疗效及新药开发有重要的指导意义