| Objective : To invigorate the mechanism of Huoxue Tongmai granule on Carotid atherosclerosis,( Carotid Arteriosclerosis, CAS) rabbit pathologic tissue structure, blood lipids TC, TG, HDL - C, LDL - C determination, interleukin - 6 (IL 6) and intercellular adhesion molecule - 1 (ICAM 1), monocyte chemotactic protein 1 (MCP 1) and nuclear transcription factor (NF-κ B) expression of intervention, and TLR4 cell signal transduction pathway as the breakthrough point, Huoxue Tongmai granule intervention rabbit Carotid atherosclerosis model the effect of rabbit plaque formation and inhibition of the expression of inflammatory markers. Methods : 1. Modeling and Grouping: In the 75 New Zealand white rabbit, randomly selected 10 for the normal group, the remaining 65 rabbit were established by the high fat feed (78% of base feed add 7% lard, 14% egg yolk powder, 1% cholesterol) carotid atherosclerosis model is set up. After 11 weeks, the models were randomly divided into the model group,the natural healing group,the Huoxue Tongmai Granule low, medium and high dose group with 7 rabbits in each group. 2. Drug Administration: Natural healing group and Huoxue Tongmai Granule low, medium and high dose group, normal feed simvastatin group 5 basic feed, invigorate the circulation through vein granule low dose group of crude drugs of 5.5 g/kg every day to fill the stomach, blood activating flux pulse particles dose groups of crude drugs in 11 g/kg every day to fill the stomach, invigorate the circulation of crude drugs through vein granule high-dose group 22 g/kg every day to fill the stomach, simvastatin group (1.87 mg / 1 kg) to fill the stomach, the natural healing group such as volume of distilled water to fill the stomach, gastric five weeks in a row. 3. Indexes Testing: In 11 weeks experiment, and week 16 full automatic biochemical analyzer determination of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL - C), low density lipoprotein cholesterol (LDL - C); Carotid artery samples, the conventional paraffin embedding slices, HE dye. Then watch pathological changes; SABC method is used to detect the expression of TLR4 in atherosclerotic plaque;Meanwhlile,ELISA method for detection of ICAM – 1and IL – 6 expression were dectected by the ELISA method; By real-time fluorescent quantitative PCR method to detect TLR4, the nf-kappa B, MCP - 1 mrna expression. Results : (1) Cholesterol targets detection: Model group TC, TG, LDL - C compared with normal group were significantly increased (P < 0.01). For five weeks, compared with natural healing group, Huoxue Tongmai granule low, medium and high dose group and simvastatin group were significantly decreased (P < 0.05); With ordinary feed Huoxue Tongmai granule low dose group, normal feed grains of blood activating flux pulse, high dose group and simvastatin group significantly decreased (P < 0.05); Ordinary feed Huoxue Tongmai granule high-dose group compared with the ordinary feed simvastatin has no statistical significance (P > 0.05). (2) Pathological observation: Normal group of carotid artery endothelial and smooth, not seen plaques and lipid, model group endothelial proliferation obviously, plaque contains a large number of foam cells, atheromatous plaque are widely distributed. Ordinary feed Huoxue Tongmai granule low, medium and high dose group compared with model group endothelial proliferation significantly reduce, plaques contain less foam cells. Ordinary feed high dose group of the arterial wall layers is clear. (3) TLR4 immunohistochemical staining: Lens, the model group than in blank group of cell plasma/membrane appeared a lot of tan calm and prompt TLR4 has a strong positive expression. Between groups compared with the natural healing group after treatment (P < 0.01). (4) ELISA method is applied to detection: Compared with the natural healing group Huoxue Tongmai granule high dose group decreased obviously (P < 0.05),compared with Huoxue Tongmai granule low-dose unclogged arteries blood through vein granule high-dose group, simvastatin group significantly decreased (P < 0.05). (5) Application of fluorescence quantitative PCR detection: After treatment groups of TLR4, the nf-kappa B, MCP - 1 mrna expression is lower than the natural healing group (P < 0.05). Conclusion : Huoxue Tongmai granule can reduce total cholesterol, triglyceride and low density lipoprotein cholesterol, may inhibit ICAM - 1, IL - 6 expression level and intervention for the formation of atherosclerosis. Huoxue Tongmai granule Added strength phlegm, invigorate the circulation of CAS plaque inhibition pulse method. By increasing dosage, thereby strengthening the delay plaque formation, stability and better achieve the goal of treatment of CAS. By increasing dosage, thereby strengthening the delay plaque formation, stability and better achieve the goal of treatment of CAS. Huoxue Tongmai granule may by TLR4 signaling pathways intervention CAS model the effect of rabbit plaque formation and inhibition of the expression of inflammatory markers. |
PDF Full Text Request