[Effects of intravenous transplantation of human umbilical cord blood mononuclear cells combined compound Danshen dripping pills on the microenvironment and apoptosis in the myocardium of the rabbits with acute myocardial infarction].

OBJECTIVE: To explore the effects of compound Danshen dripping pills (CDDP) and CDDP combined with transplantation of human umbilical cord blood cells (HUMNCs) on the inflammatory response, oxidative stress, myocardial cell apoptosis and cardiac function, and also to investigate the possible mechanisms of the combined therapy in the acute myocardial infarction (AMI). METHODS: Rabbit model of AMI successfully established by ligation of the left anterior coronary artery (LAD). Forty rabbits were randomly divided into 4 groups (n=10 per group): a control group, injected with 0.5 mL of saline in 24 h after AMI and then gavaged with 5 mL of saline daily; a CDDP group, injected with saline 0.5 mL after AMI and then gavaged with CDDP (270 mg/d) daily; a transplantation group, injected with 0.5 mL of saline contained 3 × 10(7) HUCBMCs [labeled with green fluorescent protein (GFP)] and then gavaged with 5 mL of saline daily; a combined group, injected with 0.5 mL of saline contained 3 × 10(7) HUCBMCs (labeled with GFP) and then gavaged with CDDP (270 mg/d) daily. Cardiac function index such as left ventricular fractional shorting (LVFS) and ejection fraction(LVEF) were measured by echocardiography; the pathological changes were observed by HE staining and the white blood cells in the myocardium were determined by light microscopy. The superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in myocardium were detected by nitrotetrazolium blue chloride (NBT) and thiobarbituric acid colorimetric measurement respectively. The number of transplanted cells in the myocardium was examined by GFP positive cells counted with fluorescence microscopy. RESULTS: 1) Compared with the control group (at 1 or 4 week), LVEF and LVFS were significant improved in the CDDP group, the transplantation group and the combined groups (all P<0.05), the improvement degree of cardiac function in the combined group was the most significance. There was no significant difference between the CDDP group and the transplantation group. 2) Compared with the control group (at 1 or 4 week), the number of white blood cell, myocardial cell apoptosis ratio were decreased significantly in the CDDP group, the transplantation group and the combined groups (all P<0.05), this decrease in the combined group was the most significance, and there was no significant difference between the CDDP group and the transplantation group. 3) Compared with control (at 4 week), the SOD activity was increased significantly, and MDA content in myocardium was decreased in the CDDP group, this change in the combined group was the most significance. 4) GFP-positive cells were found to be present in the peri-myocardial infarction area in the transplantation group and the combined group at 1, 4 weeks post-transplantation. The number of the GFP positive cells in the combined group was more than that in the transplantation group (P<0.05). CONCLUSION: The intravenous transplantation of HUMNCs combined with the CDDP in the treatment of rabbits with AMI could increase the survival rate of transplanted cells and inhibit the myocardial cell apoptosis, therefore improve the heart function. The possible mechanism of the combined treatment may be involved in the inhibition of the inflammatory response and oxidative stress in the myocardium following AMI.

[Effect of compound danshen dripping pill combined with intravenous transplantation of human umbilical cord blood mononuclear cells on local inflammatory response in the myocardium of rabbits with acute myocardial infarction].

OBJECTIVE: To investigate effect of Compound Danshen Dripping Pill (CDDP) on the inflammatory response of the myocardium of acute myocardial infarction (AMI) rabbits, to observe the therapeutic effect of CDDP combined intravenous transplantation of human umbilical cord blood mononuclear cells (HUCBMCs) on inflammatory response, pro-inflammatory cytokine tumor necrosis factor alpha (TNF-alpha) , and heart function in the myocardium of AMI rabbits, and to explore the possible protective mechanisms of the combined therapy. METHODS: The AMI model was successfully established by ligation of the left anterior coronary artery (LAD) in 40 healthy rabbits.Then they were randomly divided into four groups, i.e., the control group, the CDDP group, the transplantation group, and the combined group, 10 in each group. Rabbits in the control group received intravenous injection of 0.5 mL normal saline via ear vein within 24 h after AMI and then intragastric infusion of normal saline at 5 mL per day. Rabbits in the CDDP group received intravenous injection of 0.5 mL normal saline via ear vein within 24 h after AMI and then intragastric infusion of solution obtained by solving 270 mg CDDP in 5 mL normal saline per day. Rabbits in the transplantation group received intravenous injection of 0.5 mL normal saline labeled with green fluorescent protein (GFP) containing 3 x 10(7) of HUCBMCs via ear vein within 24 h after AMI and then intragastric infusion of normal saline at 5 mL per day. Rabbits in the combined group received intravenous injection of 0.5 mL normal saline labeled with GFP containing 3 x 10(7) of HUCBMCs via ear vein within 24 h after AMI and then intragastric infusion of solution obtained by solving 270 mg CDDP in 5 mL normal saline per day. At week 1 and 4 after treatment, cardiac function indices such as left ventricular fractional shorting (LVFS) and left ventricular ejection fraction (LVEF) were performed by echocardiography; the number of transplanted cells in the myocardium was found by GFP positive cells counted with fluorescence microscopy.The white blood cells in the myocardium stained with HE were determined by light microscope. The expressions of TNF-alpha protein in the myocardium were detected by immunohistochemical assay. RESULTS: (1) Compared with the control group at week 1 and 4 after treatment, the LVEF and LVFS were significantly improved in the CDDP, transplantation, and combined groups (P < 0.05). The cardiac function was significantly improved in the combined group than in the CDDP group and the transplantation group (P < 0.05). But there was no statistical difference in the latter two groups. (2) Compared with the control group, the number of white blood cells and the expression of TNF-alpha protein decreased significantly in the CDDP, transplantation, and combined groups at week 1 and 4 respectively after treatment. The number of white blood cells and expressions of TNF-alpha protein were significantly lower in the combined group than in the CDDP group and the transplantation group (P <0.05). But there was no statistical difference in the latter two groups. (3) GFP-positive cells were found to be distributed in the peri-myocardial infarction area in the transplantation group and the combined group at week 1 and 4 after transplantation. Besides, the number of the GFP positive cells was much more in the combined group than in the transplantation group (P < 0.05). CONCLUSIONS: The findings indicated that the combination of CDDP with intravenous transplantation of HUCBMCs in the treatment of AMI rabbits could elevate the survival rate of transplanted cells, and further improve the heart function. The possible mechanisms might be related to attenuating local inflammation of myocardium, and inhibiting enhanced expressions of pro-inflammatory cytokine TNF-alpha protein.