Rhodiola And Heart-protecting Musk Pill On The Rabbit Atherosclerotic Plaque Stability And Mechanism Study

Objective: 1. To get acquainted with establishment of rabbit arteriosclerotic model through high-fat diet and femoral artery angioplasty operation. To observe morphology of arteriosclerotic plaque so as to analyze its component such as foam cells,extracellular cholesterol crystal,vascular smooth muscle cells,and collagen.2. To investigate effects of simvastatin, Rhodiola rosea, She Xiang Bao Xin Wan(SXBXW) ,and Salidroside(all as angiogenesis inducing agents) on angiogenesis, extracellular matrix,and inflammation response in the arteriosclerotic plaque . Blood lipid changes are also examined.3. To compare effects of Rhodiola rosea and Salidroside on angiogenesis ,extracellular matrix,and inflammation response in the arteriosclerotic plaque and blood lipid changes. To try finding a way of standardizition and quantification of usage of Rhodiola rosea.Methods: 1. Animal models: 50 Newsealand Rabbits were feed with high-fat diet for 2 weeks. Then femoral artery angioplasty operation were performed. 2. Grouping and medicine administration: All rabbits were divided into: control group ,simvastatin group, Rhodiola rosea group, SXBXW group ,and Salidroside group.All rabbits were fed high-fat diet for another 4 weeks. Rabbits of intervention groups were fed together with medicines for 4 weeks. The dose of Rhodiola rosea (decoction with concentration of 0.5g/ml) intragastric administration was 1 ml/kg everyday; The intragastric dose of Salidroside suspension was also 1 ml/kg everyday(by mixing 1% salidroside powder with distilled water with the same concentration of salidroside as in the Rhodiola rosea decoction as 0.173mg/g) ; rabbits of SXBXW group were given 1 pills everyday;simvastatin pills were scrunched and made suspension with the administration dose of 2.5mg/kg everyday.3. Blood lipid examination:Blood total cholesterol,LDL-C,HDL,and triglyceride were determined in 1^st week,3^rd week,and 7^th week.4. Serum MMP-3,IL-6,and hsCRP were determined in 7^th week by ELISA.5. CD34,VEGF,MMP-3 ,collagen ,PCNA,MCP-1,chymase,and α SMA positive area in the arteriosclerotic plaque were analyzed and quantification by means of immunohistochemistry techniques. 6.VEGF mRNA in arterosclerotic plaque were examed by realtime PCR.7. The results were express with X|- ± S, and the difference among the groups was tested by q test (multiple comparisons of one-factor analysis of variance). SPSS10.0 was used in the data processing.Results: 1 General obersvation and description of arteriosclerotic plaque model: 10 rabbits died during the 7 weeks experiment. At the end of the experiment,the number of rabbits of each group was: control group-8 ,simvastatin group-9, Rhodiola rosea group-6, SXBXW group-8 ,and Salidroside group-9. Big grey plaques could be seen in the femoral artery and foam cells,extracellular cholesterol crystal,vascular smooth muscle cells,and collagen could be found under microscope.2 Angiogenesis in the arteriosclerotic plaque: CD34 positive areas in arteriosclerotic plaques of All medicine intervention groups were decreased(simvastatin vs cotrol:15.24±3.34% vs 20.39 ± 5.96%, P<0.01; Rhodiola rosea vs cotrol: 10.99 ± 3.59% vs 20.39 ± 5.96%, P<0.01; SXBXW vs cotrol: 13.85 ± 2.97% vs 20.39±5.96%, P<0.01; Salidroside vs control: 13.28 ± 4.01% vs 20.39 ± 5.96%, P<0.01) . VEGF positive areas in arteriosclerotic plaques of all medicine intervention groups were also decreased (simvastatin vs cotrol: 13.35 ± 7.27% vs 20.14 ± 6.15%, P<0.01; Rhodiola rosea vs cotrol: 11.43± 3.41% vs 20.14 ± 6.15%, P<0.01; SXBXW vs cotrol: 15.23 ± 2.93% vs 20.14 ± 6.15%, P<0.01; Salidroside vs control 13.59 ± 4.74% vs 20.14 ± 6.15%, P<0.01) . Rhodiola rosea and Salidroside group didn't have significant difference on those two results.3. Changes of extracellular matrix in the arteriosclerotic plaque: Collagen positive areas in arteriosclerotic plaques of All medicine intervention groups were decreased (simvastatin vs cotrol: 12.54 ± 2.82% vs 20.02 ± 6.11%, P<0.01; Rhodiola rosea vs cotrol: 16.99 ± 6.00% vs 20.02 ± 6.11%, P<0.05; SXBXW vs cotrol: 11.35 ± 1.87% vs 20.02 ± 6.11%, P<0.01; Salidroside vs control 15.65 ± 1.97% vs 20.02 ± 6.11, P<0.01) . MMP-3 positive areas in arteriosclerotic plaques of all medicine intervention groups were also decreased(simvastatin vs cotrol: 11.24 ± 2.81% vs 16.30 ± 5.96%, P<0.01; Rhodiola rosea vs cotrol: 11.89 ± 4.29% vs 16.30 ± 5.96%, P<0.01; SXBXW vs cotrol: 11.52 ± 4.97% vs 16.30 ± 5.96%, P<0.01; Salidroside vs control: 10.51 ± 2.34% vs 16.30 ± 5.96%, P<0.01) . Rhodiola rosea and Salidroside group didn't have significant difference on those two results, α SMA positive areas in arteriosclerotic plaque of Simvastatin group and SXBXW group were similar with that of control group. However, Rhodiola rosea and Salidroside could both decease α SMA positive areas (Rhodiola rosea vs cotrol: 10.34±1.62% vs 14.44 ± 2.76%, P<0.01; Salidroside vs control: 11.94±2.84% vs 14.44 ± 2.76%, P<0.01) ,and they had significant difference(P<0.05) .Simvastatin and Salidroside could decrease PCNA positive area in the arteriosclerotic plaque (simvastatin vs cotrol: 8.86 ± 5.60% vs15.07 ± 5.66%, P<0.01; Salidroside vs control 9.42±4.16% vs 15.07 ± 5.66%, P<0.01) ,but Rhodiola rosea and SXBXW could not. The difference between Rhodiola rosea group and Salidroside group was signifibcant (P<0.05) . Two medicines could reduce serum MMP-3 level(simvastatin vs cotrol: 8.38 ± 19.44 mg/L vs 21.01 ± 17.22 mg/L, P<0.01; Rhodiola rosea vs control: 12.43 ± 9.32 mg/L vs 21.01 ± 17.22 mg/L, P<0.05 ) . The difference between Rhodiola rosea group and Salidroside group was signifibcant (P<0.05) .4. Inflammation response in the arteriosclerotic plaque: All medicines could decrease chymase positive areas in the arteriosclerotic plaque (simvastatin vs cotrol: 9.74±2.11% vs 12.53 ± 2.78%, P<0.01; Rhodiola rosea vs control: 9.13 ± 1.95% vs 12.53 ± 2.78%, P<0.01; SXBXW vs control: 6.23 ± 3.71% vs 12.53±2.78%, P<0.01; Salidroside vs control: 5.57±2.13% vs 12.53±2.78%, P<0.01) . Salidroside could decrease MCP-1 positive areas (8.05±4.14% vs 12.21 ±2.59%, P<0.01), but the other medicines could not. P<0.01) .Salidroside and Rhodiola rosea group had significant difference on those two results (P<0.01) . All medicines couldn't reduce serum IL-6 and CRP signicantly.5. Blood lipid: Simvastatin could reduce serum total cholesterol ,LDL-C,HDL and triglyceride significantly. Rhodiola rosea and SXBXW could reduce LDL-C(SXBXW vs control: 6.19±14.34 mmol/L vs 12.88 ± 9.82 mmol/L, P<0.01; Rhodiola rosea vs control :8.87±4.00 mmol/L vs 12.88 ± 9.82 mmol/L, P<0.05 )),and Rhodiola rosea could further reduce total cholesterol (10.11±3.59 mmol/L vs 20.32 ±6.33 mmol/L, P<0.01) . Salidroside had little effect on blood lipid and had significant difference on serum total cholesterol with Rhodiola rosea (19.34±11.44 vs 10.11±3.59 mmol/L, P<0.01) .Conclusions: 1. Rabbit arteriosclerotic plaque model establishe with high-fat diet and femoral artery angioplasty operation contained main cells and matrix components of human lesions.2. Rhodiola rosea and SXBXW could both decrease VEGF,MMP-3, and chymase in the arteriosclerotic plaque,inhibit angiogenesis in the plaque,and reduce serum LDL-C. Those mechanisms might benefit the stability of plaque.3. The effects of inhibition of angiogenesis and production of collagen,decrease of MMP-3 and VEGF in the arteriosclerotic plaque,and reduction of serum LDL-C by Rhodiola rosea may be attributed to salidroside . Rhodiola rosea might have other important components with functions such as increasing MCP-1 and chymase in the arteriosclerotic plaque, affecting proliferation of VSMCs andreducing serum VLDL-C.