Aqueous Date Flesh or Pits Extract Attenuates Liver Fibrosis via Suppression of Hepatic Stellate Cell Activation and Reduction of Inflammatory Cytokines, Transforming Growth Factor-β1 and Angiogenic Markers in Carbon Tetrachloride-Intoxicated Rats

Figure 6

(a) Effect of date flesh extract (DFE), date pits extract (DPE), coffee and the combination groups on hydroxyproline content in hepatic tissue of CCl₄-intoxicated rats. Values are expressed as mean ± SEM. a: significantly different from normal control group; b: significantly different from CCl₄-treated group. , , . (b) Light microscopic photomicrographs of liver tissue stained with Masson’s trichrome stain (scale bar = 50 μm). (A) Control liver showing normal hepatic fibrous tissue distribution restricted mainly to the portal area, while demarcation between classic hepatic lobule is observed due to very delicate fibrous tissue. (B) Represent liver section of rat receiving CCl₄ showing marked increase of fibrous tissue arranged in irregular way between degenerated cells and sinusoids, causing destruction of classic architecture of hepatic lobules. Liver sections from rats treated with coffee (C), DFE (D), DPE (E), and the combination of coffee + DFE (F) and coffee + DPE (G) showing marked decrease of abnormally extra deposited fibrous tissue. The improvement is prominent in DFE than coffee alone or DPE alone. Sections from rats receiving combination showed apparently normal amount and distribution of fibrous tissue. (c) Effect of date flesh extract (DFE), date pits extract (DPE), coffee, and the combination groups on hepatic levels of transforming growth factor beta-1 (TGF-β1) in CCl₄-intoxicated rats. Values are expressed as mean ± SEM. a: significantly different from normal control group; b: significantly different from CCl₄-treated group; c: significantly different from coffee-treated group. , , . (d) Light microscopic photomicrographs of liver tissue immunostained with primary anti-TGF-β1 antibody (scale bar = 50 μm). (A) Control liver showing normal very weak immune reactivity of hepatocytes cytoplasm while the nuclei are not stained, and also the endothelial cells of both central vein and hepatic sinusoids are not stained. (B) represents liver section of rat receiving CCl₄ showing strong abnormal immune reactivity of most of hepatocytes’ cytoplasm and nuclei, and also most of cells of structures of portal area show strong nuclear immune reactivity. Panels (C), (D), (E), (F), and (G) represent liver sections from rat receiving coffee, DFE, DPE, coffee + DFE, and coffee + DPE, respectively, showing few hepatocytes with strong ((C), (E)) or moderate (D) immune reactivity especially those around portal area. In (F) and (G) hepatocytes’ cytoplasm and few cells of structures of portal areas show weak positivity especially in rats receiving a combination of coffee + DFE (F). (e) Light microscopic photomicrographs of liver tissue immunostained with primary anti-α-SMA antibody (scale bar = 50 μm). (A) Control liver showing normal positive immune reactivity of smooth muscle of the blood vessels of the portal area without immune staining positivity in between hepatocytes, while (B) represents liver section of rat receiving CCl₄ in which strong abnormal distributed immune reactivity, especially around degenerated hepatocytes, is prominent. Panels (C), (D), (E), (F), and (G) represent liver sections from rat receiving coffee, DFE, DPE, coffee + DFE, and coffee + DPE, respectively, showing marked decrease of the immune reactivity outside portal areas especially in groups receiving DFE and the combination.