Transcriptional activity of PPARγ in primary human adipocytes

Sammanfattning: The prevalence of obesity is increasing in most parts of the world and is a strong risk factor for the development of insulin resistance, type 2 diabetes and hypertension. Adipose tissue is mainly composed of adipocytes which store energy in the form of triglycerides and release it as free fatty acids. Adipose tissue is one of the major regulators of energy homeostasis in the body. Adipose tissue in different regions of the body has different characteristics and adipocytes in intra-abdominal fat depots are more associated with insulin resistance than adipocytes from subcutaneous fat depots.Research performed during the past several years has led to an explosion in the understanding of adipose tissue and the active role that it plays in aspects of physiology and pathophysiology. One important discovery has been identification of the nuclear hormone receptor called peroxisome proliferator-activated receptor γ (PPARγ). Peroxisome proliferator-activated receptor γ (PPARγ) is a transcription factor, which is highly expressed in adipocytes. PPARγ has been shown to affect several genes of importance for lipid metabolism, differentiation of fat cells and insulin sensitivity. The PPARγ receptor can be activated by thiazolidinediones (TZD), a class of insulinsensitising drugs, which promote fatty acid storage in fat depots and decrease glucose levels in plasma, thus, demonstrating the importance of PPARγ activity in insulin resistance and metabolic syndrome.This thesis has investigated the transcriptional activity of PPARγ in a clinically relevant cell type for insulin resistance and type 2 diabetes; the primary human adipocyte. For this purpose, a method for transfection of primary human adipocytes by electroporation and for measurement of the activity of PPARγ has been developed and optimised. This method has been used to study the effect of saturated and unsaturated fatty acids on the transcriptional activity of PPARγ. Interestingly, it was been found that saturated fatty acids can activate PPARγ, thus promoting a protection against diabetes. The strongest activator was the monounsaturated palmitoleic acid. The transcriptional activity of PPARγ in primary human adipocytes from intra-abdominal and subcutaneous adipose tissues was also examined. It was found that PPARγ activity is considerably lower in adipocytes from visceral compared with subcutaneous fat from the same subject. Another reason for using human tissue to reach clinical relevance shown here was that the same difference in PPARγ activity could not be found between intra-abdominal and subcutaneous fat tissues in mice. This finding may serve as the basis of why excess intraabdominal fat tissue is associated with high risk for development of type 2 diabetes and cardiovascular diseases.The blood pressure regulating renin-angiotensin system (RAS) in human adipose tissue and in isolated adipocytes was examined and related to PPARγ. It was found that the production of angiotensin II, which is an important hormone for increasing the blood pressure, can be produced by isolated adipocytes and that the production is higher in adipocytes coming from omental than subcutaneous fat tissue. Further, it was shown that angiotensin II inhibits PPARγ activity in omental adipocytes, thus reducing the insulin sensitivity. Therefore, this study connects two of the major risk factors in obesity; diabetes and hypertension, and may also explain how drugs, which inhibit the RAS, can also be protective against diabetes. In conclusion, the findings in this thesis give new knowledge about regulating mechanisms of fat cells and its importance in diabetes and cardiovascular disease.