| Objective:To investigate the lipid metabolic, oxidative, endothelial dysfunction and anti-atherosclerosis effects of hirudo and its possible mechanism in animal model of experimental atherosclerosis.Method:32healthy male Sprague-Dawlay rats were randomly divided into four groups:normal control group (N-ctrl); model control group (M-ctrl); hirudo low dose group (H-low); hirudo high dose group (H-high). In the beginning, all groups were adaptive fed by normal diet for one week. The N-ctrl group was fed with normal diet, the other groups were intraperitoneal injected by6×105unit vitamin D3in the third day, and corresponding medicines were intragastric administration every day on the basis of atherogenic diet for10successive weeks. At the same time, normal saline was given to the rats of N-ctrl group and M-ctrl group with the same volume and the same way. The weekly body weight of rats was recorded. After ten weeks, all rats were killed. The liver, epididymal and perirenal adipose tissue were collected and weighed. Levels of blood lipid profile, homocysteine (HCY), catalase (CAT), plasma fibrinogen (FIB) were observed. The activity of liver lipoprotein lipase (LPL) and hepatic lipase (HL) were observed. The expression of oxidized low-density lipoprotein (ox-LDL) in serum and lectin like oxidized low density lipoprotein receptor-1(LOX-1) in aortic wall were detected by ELISA. Hematoxylin-eosin and oil red O stain were usd for morphological investigation by light microscope.Results:1. In the beginning of experiment, the body weight of rats was no difference in all groups. The weekly body weight of rats was increased in all groups. Since the experiment second week, the weight of rats in the M-ctrl group was increased compared with the N-ctrl group (P<0.05), Compared with the M-ctrl group, the body weight of rats was decreased in the H-low and H-high groups (P>0.05); In the M-ctrl group, the weight of epididymal and perirenal adipose were increased compared with the N-ctrl group (P<0.01or P<0.05), Compared with the M-ctrl group, the weight of epididymal and perirenal adipose were decreased in the H-low and H-high groups (P<0.05). 2. In the M-ctrl group, the serum TG, TC, LDL-C levels were increased and the HDL-C level was decreased compared with the N-ctrl group (P<0.05or P<0.01). Compared with the M-crtl group, the serum TC level was decreased (P>0.05), the serum TG and LDL-C levels were decreased and the HDL-C level was increased in the hirudo group (P<0.05or P<0.01). Compared with the H-low group, the TC, TG, LDL-C levels were decreased and the HDL-C level was increased in H-high group (P>0.05).3. In the M-ctrl group, The activity of liver LPL and HL were decreased (P<0.05or P<0.01) compared with the N-ctrl group. Compared with the M-ctrl group, The activity of liver LPL and HL were increased in the hirudo groups (P<0.05), Compared with the H-low group, The activity of liver LPL and HL were increased in H-high group (P>0.05).4. In the M-ctrl group, the plasma FIB level was increased (P<0.05) compared with the N-ctrl group. Compared with the M-ctrl group, the plasma FIB level was decreased in the hirudo groups (P<0.05), Compared with the H-low group, the plasma FIB level was decreased in H-high groups (P>0.05).5. In the M-ctrl group, the serum HCY level was increased (P<0.05) and the CAT activity was decreased (P<0.01) compared with the N-ctrl group. Compared with the M-ctrl group, the HCY level was decreased (P<0.01) and the CAT activity was increased (P<0.05) in the hirudo groups. Compared with the H-low group, the HCY level was decreased (P<0.05) and the CAT activity was increased (P>0.05) in H-high groups.6. In the M-ctrl group, the expression of ox-LDL in serum and LOX-1in aorta were reinforced compared with the N-ctrl group (P<0.05or P<0.01). Compared with the M-ctrl group, the the expression of ox-LDL in serum and LOX-1in aorta were attenuated in the hirudo groups (P<0.05or P<0.01). Compared with the H-low group, the expression of ox-LDL was attenuated in the H-high group (P<0.05), the expression of LOX-1was attenuated (P>0.05) in H-high groups.7. Pathomorphological changes in aorta:HE stain:In the N-ctrl group, the structure of the aortic wall was well constructed, the intima was thin and endothelia was intact. In the M-ctrl group, some endothelial cells were lost and intima thicked, the smooth muscle cells lined up in disorder, the elastic fibers were proliferated and disorder. Compared with the M-ctrl group, intima was lightly thicked, the smooth muscle cells slightly proliferated and lined up in order in the hirudo group. Oil red O stain:In the N-ctrl group, no lipid droplet deposit in the aorta. In the M-ctrl group, some lipid droplet deposit in the aorta. Compared with the M-ctrl group, the amount of lipid droplet deposition was reduced in the hirudo group rats arota.Conclusions:The rat model of early atherosclerosis can be established successfully by atherogenic diet combined with intraperitoneal inject vitamin D3is mimic to human atherosclerotic disease and suitable for the observation of the smooth muscle cells lined up in disorder and the elastic fibers proliferates.(1) The results suggest that the hirudo can ameliorate the atherosclerotic changes and its protection effect may be involve in decreasing body weight and fat deposition.(2) Hirudo has effect on decreasing the serum level of TC, TG, LDL-C and increasing the serum level of HDL-C increasing the The activity of liver LPL and HL in AS rats. Regulating serum lipid metabolic disorder, preventing cholesterol accumulation.(3) Hirudo has obvious effect on decreasing the serum level of HCY and reinforcing the activity of CAT in AS rats, enhancing anti-oxidant capacity, alleviating oxidative damage.(4) Hirudo has obvious effect on attenuating the expression of ox-LDL and LOX-lin AS rats, restraining ox-LDL-mediate cell apoptosis, necrosis and autophagic cell death.(5) Between blood lipid profile and the oxidative modification lipoprotein and lipid metabolic enzymes have certain correlation. |
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