[3](#Fig3){ref-type="fig"}a-f).Fig. 3The expression of Mmps and collagens was analyzed in adipose tissue from HFD mice and WT mice. **a**--**c** qRT-PCR analysis showed significantly increased expression of Mmps, as well as collagens (Col) III, IV, and V in adipose tissue from HFD mice compared with WT mice. **d**--**f** Increased expression of Col IV in adipose tissue from HFD mice compared with WT mice was confirmed by Western blotting. **g**--**i** Representative IHC staining results of Mmp2 in adipose tissue from WT mice or HFD mice. Scale bar: 50 μm
In addition, IHC analysis of Mmp2 in adipose tissue from WT mice or HFD mice showed positive staining mainly in the cytoplasm and matrix of adipocytes (Fig. [3](#Fig3){ref-type="fig"}g). Mmp2 expression was also detected by Western blot analysis in adipose tissue from WT mice or HFD mice, and the band density of Mmp2 from the HFD mice was significantly higher than that of the WT mice (Fig. [3](#Fig3){ref-type="fig"}h, i).
The Effect of Obese status on Macrophage in Adipose Tissue {#Sec15}
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To further understand the role of macrophages in the adipose tissue of obese mice, the expression of M1 marker gene iNOS was examined in epididymal adipose tissue and subcutaneous adipose tissue in obese and lean mice. As shown in Fig. [4](#Fig4){ref-type="fig"}a, iNOS expression was higher in epididymal adipose tissue in obese mice than in lean mice. In subcutaneous adipose tissue from obese mice, we found significant iNOS expression compared with lean mice.Fig. 4The effect of obesity on macrophage infiltration in adipose tissue. **a** Immunohistochemical analysis of iNOS expression in epididymal adipose tissue and subcutaneous adipose tissue from WT mice and HFD mice. Scale bar: 50 μm. **b** The effect of the HFD and insulin treatment on the body weight and Lee index of H
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