Abstract

Aim of the study

The inhibitive effect of BuYang HuanWu Decoction (BYHWD) and its major components on vascular intimal hyperplasia and the expressions of cell cycle protein and extracellular matrix protein.

Materials and methods

Sprague-Dawley rats were randomly divided into sham-operated, control, alkaloid, glycoside, BYHWD and atorvastatin groups. Rat aorta intima in all groups were injured by insesion of domestic balloon catheter into the aortae except sham-operated rats. Drugs were administrated orally from the second day after vascular injury and continued for 14 days. The injured segments of aortae were collected on the sixteenth day after operation to observe the morphological changes of vascular structure and to examine the expressions of proteins in vascular cells associated with cell cycle including proliferating cell nuclear antigen(PCNA), cyclinD₁ and cyclinE, and extracellular matrix(ECM) proteins including collagen I (Col-I) and fibronectin (FN), further to discover the involved biologically active substances and the potential mechanisms.

Results

Alkaloid and glycosid isolated from BYHWD were more effective than BYHWD in the inhibition of intimal hyperplasia and the expressions of PCNA, cyclinD₁, cyclinE, Col-I and FN, suggesting that alkaloid and glycoside may be the main components of BYHWD responsible for the observed inhibition of excessive hyperplasia of vascular intima.

Conclusions

The mechanism associated with the anti-hyperplasia activity of BYHWD and its effective components may be related to the blockage of cell cycles of VSMC, and the inhibition of the ECM protein synthesis, even the increased degradation of ECM proteins.

Keywords: BuYang HuanWu Decoction; Atorvastatin; Vascular intimal hyperplasia; Vascular smooth muscle cell; Morphology; Proliferating cell nuclear antigen; CyclinD₁; CyclinE; Extracellular matrix; Collagen I; Fibronectin

Abbreviations: BYHWD, BuYang HuanWu Decoction; CDK, cyclin-dependent kinase; Col-I, collagenI; ECM, extracellular matrix; FN, fibronectin; HPLC, high performance liquid chromatograph; PCNA, proliferating cell nuclear antigen; PTCA, percutaneous transluminal coronary angioplasty; TCM, traditional Chinese medicine; VSMC, vascular smooth muscle cell

Article Outline

1. Introduction

2. Materials

2.1. Experimental animals

2.2. Balloon catheters

2.3. Composition identification, vouchers specimens, active ingredients detection and the positive control drug option of BYHWD

2.3.1. Composition identification and vouchers specimens

2.3.2. Standard preparation and control drug

2.3.3. The basis on choosing the active ingredients

2.3.4. The option of positive control drug

2.4. Reagents

2.5. Apparatus

3. Methods

3.1. The water-extract preparation of BYHWD

3.2. Extraction and separation of all effective components of BYHWD

3.3. Qualitative and quantitative analysis of active principles

3.3.1. Chemical identification

3.3.2. Content analysis

3.3.2.1. Chromatographic condition

3.3.2.2. Determination of the content of astragaloside IV from BYHWD

3.3.2.3. Determination of the content of paeoniflorin and amygdalin from BYHWD

3.3.2.4. Determination of the content of tetramethylpyrazine from BYHWD

3.4. Animal model

3.5. Grouping and administration methods

3.6. Vessel acquisition and indices determination

3.6.1. The method of vessel acquisition

3.6.2. Determination of vascular morphological indices

3.6.3. Determination of expressions of PCNA, SMα-actin, cyclinD₁, cyclinE, Col-I, and FN

4. Statistical analysis

5. Results

5.1. Comparisons of vascular morphological indices

5.2. PCNA and SMα-actin expressions in different groups

5.3. CyclinD₁ and cyclinE expressions in different groups

5.4. Col-I and FN expressions in different groups

6. Discussion

Acknowledgements

References