Protective Effects Of The Compound Injection Of Danshen On Vascular Endothelial Cells Induced By Hypoxia-reoxygenation Injury

Objective: To study the protective effects of the compound injection of Danshen on cultured vascular endothelial cells (VEC) induced by hypoxia-reaxygenation injury , nitric oxide (NO) and total-antioxidation capacity (T-AOC) produced and intercellular adhesion molecule-1 (ICAM-1) by endothelial cells, and to further study the Danshen for mechanisms of anti -atherosclerosis.Methods: By using primary culture of human umbilical vein endothelial cells (HUVEC) in vitro, the VECs were divided into three groups: (1) normal group: normal VECs, (2) hypoxia-reoxygenation group: VECs were treated with 1 mM sodium dithionite and continue to be cultured for 12 hours by adding whole M199, and (3) Danshen group: VECs were continuously cultured with different concentrations of the Danshen after hypoxia. The morphological changes of VECs in each group were observed by the inverted microscope, and the expression of ICAM-1 in each group was detected by immunocytochemistry. The optical density (OD) value of cells was detected by MTT colorimetric method, content of MDA, T-AOC and NO in the supernatant fluid of cultured cells was detected by spectrophotometer.Results: The results detected by immumocytochemistry showed that the expression of ICAM-1 was very strong in the hypoxia-reoxygenation group, but one was weak in the normal group and Danshen group. In addition, MTT assayshowed that Danshen had the protective effects for VECs induced by hypoxia-reaxygenation injury in 0.25-12.5 mg/ml of concentration. The OD value was obviously decreased in the hypoxia-reoxygenation group, but one was increased in the normal and Danshen groups and was highest at 12.5 mg/ml of Danshen concentration. The OD value had significant difference (P<0.01) between normal group (0.473 + 0.02), Danshen group (0.400 + 0.03) and hypoxia-reoxygenation group (0.183+0.02). The content of MDA and NO in the supernatant fluid of cultured cells also had significant different between the hypoxia-reoxygenation group and normal group, Danshen groups. Namely, MDA content (5.042 + 0.20 nmol/ml) was increased and NO content (25.66 + 1.35|iimol/L) was decreased in the hypoxia-reoxygenation group, in contrast, MDA content (2.00 + 0.23 nmol/ml and 2.390 + 0.13 nmol/ml) was decreased and NO contents (93.56 + 0.8lumol/L and 84.04 + 0.68 |imol/L) was increased in the normal and Danshen groups. The transudatory content of T-AOC had significant different (/><0.01) between the hypoxia-reoxygenation group (2.543 + 0.24 U/ml) and normal group (9.003 + 0.09 U/ml), Danshen groups (7.662 + 0.50 U/ml).Conclusions: The results indicate that the compound injection of Danshen has protective effects for the HUVEC induced by hypoxia-reaxygenation injury and anti-anoxic injury by the mechanisms of abating the lipid peroxidation of cell, counterpoising the content of NO and decreasing over expression of ICAM-1.