Hydroxysafflor Yellow A Suppress Liver Fibrosis Via Regulating PPAR-γ/p38 MAPK Signal Pathway

Objective:Recently, one approach to protect against carbon tetrachloride (CC14) with high fat diet (HFD)-induced liver fibrosis is the use of herb-derived natural compounds, such as Hydroxysafflor yellow A (HSYA). The antifibrosis effect of HYSA against liver fibrosis has been investigated; however, its mechanisms have not been entirely revealed.Our study will reveal the role of Peroxisome proliferator-activated receptor(PPAR-y)/p38 signal pathway in its mechanisms.Methods:1. CC14 and HFD were used to mimic liver fibrosis model. The biochemical indicators levels of AST, ALT, HA, LN, Ⅲ-C and Ⅳ-C in serum between difficent groups were determined according to the procedures described in the manufacturer’s assay kits; Hydroxyproline levels in the liver tissue between difficent groups were analyzed using alkaline hydrolysis kit.2. Liver fibrosis grades and histopathological changes were quantified by haematoxylin and eosin (H&E) or Masson’s trichrome.3.Immunohistochemical staining and enzyme-linked immunosorbent assay were used to evaluate the expression level of TGF-β1, TIMP-1, MMP-2, TNF-α in liver tissues between difficent groups.4. The expression of PPAR-y, p38, p-p38, TGF-(β1, TIMP-1 and MMP-2 in gene or protein level between difficent groups were detected by using RT-PCR or western blot.Results:1. Compared with normal group, the levels of ALT,AST,LN, HA, Ⅲ-C, Ⅳ-C in serum and the hydroxyproline content in liver tissue were significantly elevated in treatment groups. The levels of each of these parameters were further increased by treatment with PPAR-γ antagonist GW9662. Compared to the model group, this increased were attenuated in HSYA and HSYA+GW9662 groups. HSYA treatment significantly alleviated liver injury in rats.2. Control group revealed normal liver morphology. Treatment groups showed typical damage in the liver structure, including hepatic steatosis, necrotic granulomas, infiltration of inflammatory cells, and collagen deposition with pseudo-lobe formations. Compared to the model group, this damage were attenuated in HSYA and HSYA+GW9662 groups. HSYA treatment significantly alleviated liver injury in rats.3. Compared with normal group, the levels of TGF-β1, TIMP-1, TNF-α were significantly elevated in treatment groups. The levels of each of these proteins were further increased by treatment with PPAR-γ antagonist GW9662. Compared to the model group, this increased were attenuated in HSYA and HSYA+GW9662 groups. HSYA treatment significantly alleviated liver injury in rats. By contrast, the result of MMP-2 is on the contrary.4. Compared with normal group, the genes of TGF-β1, TIMP-1 were significantly elevated in treatment groups. The levels of each of these genes were further increased by treatment with PPAR-Y antagonist GW9662. Compared to the model group, this increased were attenuated in HSYA and HSYA+GW9662 groups. HSYA treatment significantly alleviated liver injury in rats. By contrast, the result of PPAR-γ, MMP-2 is on the contrary. The level of phospho-p38 MAPK was uniform with above-mentioned, since the total level of this protein was unchange in all groups.Conclusions:1. HSYA noteworthy ameliorates hepatic fibrosis induced by CCL4 with HFD.2. HSYA reduces the expression levels of pro-fibrotic factors and elevates hepatoprotective factors.3. Hydroxysafflor yellow A suppresses liver fibrosis by regulating PPAR-γ/p38 MAPK signaling...