Protective Role Of Persimmon Tannin Against Atherosclerosis In ApoE-/-mice

Objective:To investigate the protective role of persimmon tannin(PT) against atherosclerosis, and to clarify the potential anti-atherosclerotic mechanisms in Apo E-/- mice.Methods:1.Forty-five male Apo E-/- mice of aged 7 week were divided randomly into three groups of fifteen animals each as follow:(1) Normal diet group(ND)(2) High fat diet group(HFD)(3) HFD+25mg/kg·bw persimmon tannin group(HFD+PT). Fifteen wild type(WT, C57BL/6J) mice were used as the normal control group. Mice in PT group were orally administrated with 25mg/kg·bw PT, the other group were supplemented by stroke-physiological saline solution. High fat diet was comprised of 21% fat and 1.25% cholesterol. 2.During 12 weeks, body weight were recorded weekly. And at the end of 12 weeks, the aoras were prepared for H&E, Oil red O staining and transmission electron microscope observation to make sure AS model established. After 12 weeks, all mice were killed by decapitation under anesthesia and the blood samples were collected for serum lipid(TG, TC, LDL-C, HDL-C) analysis and antioxidant capacity(SOD, GSH, MDA) determanation. Besides, the ox-LDL levels, IL-1β and TNF-α contents in serum were detected by ELISA technique. The m RNA expression of STAT1, STAT6, CD36 and NOXs subunit in aorta were quantified with RT-PCR method.Aortic ROS levels were detected by fluorescent probe-DHE. The expression of ox-LDL recptor CD36, i NOS, CD206 and p47 phox in aortic arch were determined by immunohistochemistry(IHC).Result:1.Oil red O staining analysis revealed that PT supplementation decreasd AS lesion size in HFD+PT group than in HFD group(47.14% vs 23.24%, P<0.01). And PT could decrease pathological morphology and ultrastructural change. 2.Compared with HFD group, the concentration of serum TG, TC, LDL-C, ox-LDL were significantly reduced in PT group. 3.Compared wih HFD group, the level of serum IL-1β、TNF-α in HFD+PT group showed a marked decrease 33.3%, 38.01% respectively(P<0.01). 4.The PT could inhibit the expression of ROS in aorta(detected by ROS probe DHE) and MDA in serum.While increased the content of GSH and SOD. 5.Compared with ND group, the expression of NOX4, p47 phox, gp91 phox and p22 phox in HFD group were increased obviously. HFD+PT group changed the expression, which further explained the protective action of PT. 6.The m RNA and protein expression of CD36 in HFD+PT group were lower than that of PT group. 7.The m RNA expression of STAT1 on aorta and protein expression of i NOS in PT group were significantly lower than that of HFD group, while STAT6 m RNA expression and CD206 protein expression were upregulated.Conclusion:The results elucidated that PT supplementation play an important role in preventing HFD-induced AS. The effect were partially related to decrease blood lipids, reduce inflammatory cytokine, regulate the macrophage polarizationion factor and l adjust the expression of NOXs.