[Experimental study of Xuefu Zhuyu Decoction induced participation of endothelial progenitor cells in the angiogenesis of the ischemic region].

OBJECTIVE: To observe the effects of Xuefu Zhuyu Decoction (XFZYD) on the angiogenesis and injury repair in the ischemic region. METHODS: The ischemic hind limb rat model was established using Bletilla embolization. Endothelial progenitor cells (EPCs) were labeled with DAPI and injected into the model rats from the vena caudalis. Then rats were treated with different doses of XFZYD by gastrogavage. Blood was withdrawn. The granulation tissue and the muscle tissues from ischemic and necrotic portion were taken on the 3rd and 7th day of the medication. The samples were frozen and sliced to analyze the fluorescent expressions. The necrosis of each sample was observed by routine pathological section. The vessels number was counted. The serum NO levels were detected using nitrate reductase method. The macro-morphological observation of ischemic lower limbs were lasted for 30 days. RESULTS: After 3 and 7 days of medication, the fluorescence intensity of ischemic area and the number of granulation vessels were significantly more in the high dose XFZYD group than in the routine treatment group and the normal saline treatment groups. The aforesaid indices were significantly higher in the routine treatment group than in the normal saline treatment group after 7 days of medication. The serum NO concentrations were significantly lower in the normal saline group at other time points. On the 30th day of medication, the muscular atrophy of the ischemic hind limbs was the least significant in the high dose XFZYD group. CONCLUSION: XFZYD could improve the ischemic necrosis by improving the NO level, inducing the EPCs' migration to the ischemic region, and promoting the angiogenesis.

[Effects of Xuefu Zhuyu decoction on EPCs function].

OBJECTIVE: To observe the effect on EPCs function by Xuefu Zhuyu Decoction. METHODS: After induced by serial concentrations of Xuefu Zhuyu Decoction-contained serum (XZDCS) and blank serum, EPCs proliferation, migration, adhesion and uptake function were detected by MTT, Boyden chamber, adhesion and uptake of ac-LDL respectively. RESULTS: Compared with the control group, 15% and 10% XZDCS could elevate the cell regeneration for 24 h and 48 h respectively. Both concentrations could improve the EPCs migration and adhesion for all 24, 48, 72 h and uptake of ac-LDL only 48 h. But 10% XZDCS could last effect on uptake of ac-LDL to 72 h and 5% XZDCS converted the inhibition of cell adhesion in the first 24 h to promotion in the next 48 - 72 h. CONCLUSION: Xuefu Zhuyu Decoction could induce EPCs differentiation into EC to angiogenesis by promoting its proliferation, migration, adhesion and uptake function.

The effect of Xuefu Zhuyu Decoction on in vitro endothelial progenitor cell tube formation.

OBJECTIVE: To observe the effect of Xuefu Zhuyu Decoction ()-containing serum (XFZYD-CS) on endothelial progenitor cell (EPC) tube formation in vitro. METHODS: Mononuclear cells from rat bone marrow were prepared in a Ficoll density gradient centrifuge. EPCs were separated by the differential attachment method, and observed with inverted microscope for the effect of XFZYD-CS on EPC tube formation. RESULTS: After one day, EPCs exposed to the serum containing 5%, 10% and 15% XFZYD-CS formed typical tubes or vessel networks. The tube formation time was two days ahead of the control group and the size of most tubes in the serum groups was smaller than in the control group. CONCLUSION: XFZYD-CS could induce EPC angiogenesis and hasten tube formation, especially in capillary vessels. The study provides experimental evidence for the plausibility of Xuefu Zhuyu Decoction in the treatment of ischemic diseases.

[Preparation of sustained release multivesicular liposome for thymopentin and preliminary study on its pharmacokinetics in rats].

To optimize the formulation and preparation method of multivesicular liposome of thymopentin and to investigate its pharmacokinetics in rats, the multivesicular liposome of thymopentin was prepared by double emulsification method and the formulation was optimized by orthogonal design. The release characteristics of thymopentin from multivesicular liposome in PBS (pH 7.4) and in plasma were investigated. The multivesicular liposome of thymopentin labeled with fluorescein isothiocyanate was prepared by double emulsification method. Its pharmacokinetics was evaluated following intramuscular injection in rats. The optimal formulation of multivesicular liposome of thymopentin were formulated with 7.5% glucose in aqueous phase and 2.25 mol x L(-1) triolein, 2.68 mol x L(-1) DPPG and 16.96 mol x L(-1) DOPC in organic phase. The entrapment efficiency of the multivesicular liposome of thymopentin was above 85% and the mean particle size was about 22 microm. The in vitro release of thymopentin from multivesicular liposome in PBS (pH 7.4) and in plasma was found to be in a sustained manner. The release curves were fitted to Higuchi equation. The pharmacokinetics following intramuscular injection of the multivesicular liposome of thymopentin labeled with fluorescein isothiocyanate in rats showed that the peak concentration of thymopentin was lower and elimination of it was slower significantly than that of thymopentin labeled with fluorescein isothiocyanate solution in the same dose. The plasma concentration of thymopentin maintained above quantitative limitation at 120 h after administration of multivesicular liposome of thymopentin. The optimized formulation and preparation technology of multivesicular liposome of thymopentin with higher entrapment efficiency are feasible with good reproducibility. Multivesicular liposome of thymopentin showed significant sustained-release property following intramuscular injection in rats.