Research On Berberine Improves Insulin Resistance Through Inhibition Of Adipose Tissue Macrophages Infiltration

This topic is based on the latest development of etiology and pathogenesis of Traditional Chinese Medicine(TCM) "type 2 diabetes hot filled" and modern research "inflammation theory", "the theory of traditional Chinese medicine and obesity hot filled" further development of insulin resistance (IR) of type 2 diabetes and combination, put forward the TCM pathogenesis of obesity in "hot Sheng", using "the principle of treatment of heat clearing and detoxifying" therapy, to lower blood sugar in treating type 2 diabetes mellitus objective. This research selects the main constituents of berberine one commonly used micro powder of Huanglian clinical (BBR) as a representative drug research, to observe the effects of obesity related IR. Discussion on IR type 2 diabetes related "hot filled" pathogenesis, "inflammation theory" and "the principle of treatment of heat clearing and detoxifying" treatment of the connection between the hypoglycemic mechanism proposed, clearing away heat and toxic material.Objective:The diet-induced obesity (DIO) C57BL/61asc mice and cultured macrophages induced by RAW264.7 and 3T3-L1 in adipocytes, observe effect of main components of Coptis BBR on macrophages infiltration in adipose tissue and the degree of obesity related insulin sensitivity. In this study, BBR decreased body weight, reduce adipose tissue macrophage infiltration, mechanism of action of drugs in the treatment of IR, rich in traditional Chinese medicine monomer slimming hypoglycemic theory, to have a further understanding of the new role clear treatment of traditional Chinese medicine.Research methodsIn vivo experiment:1) the establishment of diet-induced obesity (DIO) C57BL/61asc mice, the sensitivity index for 18 weeks to establish animal models of obesity and evaluation of insulin, pathology was observed by HE staining of adipocytes and adipocytes diameter size measurement, at the same time with berberine (BBR), and rosiglitazone (RSG) were selected as the positive control drug intervention for 15 days study.2) by the analysis of adipose tissue macrophages by flow cytometry (ATMs) and the number of Western blot in adipose tissue of the inflammatory signaling pathway and insulin signaling pathway, the expression of the real time PCR, IL-6, TNF-alpha gene in adipose tissue.3) Using ELISA method, we detect serumMCP-lof mices.In vitro experiment:1) RAW264.7 macrophages cultured and induced by 3T3-L1 adipocytes. With glucose oxidase method to observe 10umol/L BBR macrophages regulating liquid on glucose consumption.2) The inflammatory signaling pathway JNK detected by Western blot method (Thr183/185), NF-κB/P65, P-IKKβ (ser181) phosphorylation protein.3) using trans well co culture system, the macrophages were stained with hematoxylin eosin method, observation of 3T3-L1 adipocytes on RAW264.7 macrophage chemotaxis and the number of detected with ELISA in supernatant of 3T3-L1 adipocytes MCP-1.Results1, using TNF-a as the representative of the cytokines can make fat cells can activate JNK, IKKβ signaling pathway, promoting cytokines IL-6, TNF-a and MCP-1 gene expression increased, Ser307 protein phosphorylation activation, decrease insulin sensitivity, glucose consumption decreased.2, LPS JNK, IKKβ promote the activation of NF-kB signaling pathway key protein, the cytokines IL-6, TNF-α and MCP-1 gene expression, cells showed inflammatory state.3, high fat diet for 18 weeks, the body weight of mice serum MCP-1, FBG, FINS increased significantly, HOMR Idex>1, decreased insulin sensitivity.4 berberine can inhibit the JNK、IKKβ/NF-κB signaling pathway, inhibiting inflammatory factors IL-6, TNF-a and chemokine MCP-1 gene expression, while inhibiting the infiltration of ATMs, inhibition of serine 307 site (Ser307) activated protein phosphorylation, improvement insulin resistance.ConclusionBerberine can inhibit the number of macrophages in adipose tissue and inhibit the inflammatory signaling pathway, improve insulin sensitivity.