ABSTRACT
Objective: To investigate the inhibitory effect of coenzyme Q10 (Co-QlO) on the apoptosis of human coronary endothelial cells (HCAECs) induced by high glucose, and to elucidate its possible mechanism. Methods: The HCAECs were divided into control group, high glucose group and high glucose combined with 5, 10, and 20 μmol · L-1 Co-QlO treatment groups; the HCAECs in control group were cultured for 24 h using a routine culture method. The cells in high glucose group were treated with 30 mmol · L-1 glucose for 24 h; the cells in high glucose combined with 5, 10, and 20 μmol · L-1 Co-QlO treatment groups were treated with 5, 10, and 20 jumol · L-1 Co-QlO combined with 30 mmol · L-1 glucose for 24 h, respectively. The cell viabilities of HCAECs in various groups were measured by CCK-8 assay. The apoptotic rates of HCAECs in high glucose group and high glucose combined with 10 μmol · L-1 Co-QlO treatment group were detected by Hoechst-PI double staining. The cell mitochondrial membrane potentials of HCAECs in high glucose group and high glucose combined with 10 μmol · L-1 Co-QlO treatment group were determined by Mito-tracker staining. The mitochondrial reative oxygen species (mtROS) levels in the HCAECs in high glucose group and high glucose combined with 10 jumol · L-1 Co-QlO treatment group were measured by MitoSox staining. The protein expression levels of B cell lymphoma/leukemia 2 protein (Bcl-2), Bel-2 assaciated X protein (Bax), Bel-2 assaciated death promoter (Bad) and X-linked inhibitor of apoptosis protein (x-IAP) in the HCAECs in high glucose group and high glucose combined with 10 μmol · L-1 Co-QlO treatment group were detected by Western blotting method. Results: Compared with control group, the cell viability of HCAECs in high glucose group was significantly reduced (P<0. 01); compared with high glucose group, the cell viabilities of HCAECs in high glucose combined with 5, 10, and 20 μmol · L-1 Co-QlO treatment groups were significantly increased (P<0. 05 or P<0. 01), especially in high glucose combined with 10 jumol · L-1 Co-QlO treatment group (P<0. 01). Compared with high glucose group, the apoptotic rate, the mitochondrial membrane potential and the mtROS level of HCAECs in high glucose combined with 10 μmol · L-1 Co-QlO treatment group were significantly decreased (P<0. 01). The Western blotting results showed that compared with high glucose group, the expression levels of Bax and Bad proteins in the HCAECs in high glucose combined with 10 μmol · L-1 Co-QlO treatment group were decreased significantly (P<0. 01), and the expression levels of Bcl-2 and x-IAP proteins were increased significantly (P<0.01). Conclusion: Co-QlO may reduce the apoptosis of HCAECs induced by high glucose through inhibiting the mitochondrial apoptosis-related pathway to ptotect the cells.
ABSTRACT
BACKGROUND: Cerebral infarction is commonly associated with blood stasis syndrome. Abnormal alternation of blood rheology is generally manifested as increased blood viscosity and hematocrit (HCT). In isometric hemodilution, a certain amount of red blood cell (RBC) is shifted by bleeding and simultaneously, isometric diluter is supplemented to reduce whole blood viscosity.OBJECTIVE: To observe the improvement of astragalus injection, the Chinese herb for qi tonification and isometric hemodilution on blood rheology in blood stasis syndrome of cerebral infarction.DESIGN: Randomized controlled experiment and case-control analysis were designed.SETTING: Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology. PARTICIPANTS: In cerebral infarction group (infarction group), 64 inpatients of senile ischemic cerebral vascular disease were collected from Union Hospital Affiliated to Huazhong University of Science and Technology from March 2002 to March 2004. Al l of cases were aged over 60 years and were in conformity with the diagnostic criteria on blood stasis syndrome. According to random number table, routine treatment group (routine group) and the group of integrative therapy of Chinese and western medicine (experimental group) were divided, 32 cases in each one. 47 healthy people of similar age and diagnosed with routine physical examination were selected in normal control. METHODS: In routine group, cerebral infarction was treated with routine therapy, including extending capacity, reducing viscosity, resisting coagulation, blocking aggregation of platelet and dehydration and general symp tomatic supporting treatment. In experimental group, on basis of routine treatment, isometric hemodilution and astragalus injection, the Chinese herb for qi tonification were used. 10% of total blood amount (about 450-650 mL) was collected from vein, and colloid solution of same volume was injected intravenously. The treatment was applied once every 5 days, continuously for 3 times. Astragalus injection 50 mL mixed with physical saline 250 mL was intravenous dropped, once per day, continuously for 3 weeks. MAIN OUTCOME MEASURES: ① Comparison of indexes in bloodrheology before and after treatment in routine group and experimental group. ② Comparison of indexes in blood rheology between normal control and infarction group. RESULTS: According to intention management, 64 patients and 47 normal persons all entered result analysis. ① Comparison between infarction group and normal control: RVB, HCT and PFC (fibrinogen) were higher than normal control [(3.90±0.73), (3.40±0.28) mPa·s; (46.39±6.03) %,(42.61±2.91)%; (3.25±0.75), (3.08±0.46) g/L, P < 0.01, 0.05], MTIE (de formity index of RBC) was lower than normal control (0.958±0.006, 0.961 Shen H,Lu YD.Study on quantitative messurement of immunohistochemical ±0.004, P < 0.05). ② Comparison between routine group and experimental group: Difference in some indexes presented before the treatment. After treatment, RVB, HCT and PFC in experimental grou p were all lower than routine group [(3.90±0.52), (4.21±0.68) mPa·s; (43.80±3.29)%, (48.47±4.50)%; (3.31±0.60), (3.68±0.67) g/L, P < 0.01, 0.05]. CONCLUSION: Isometric hemodilution therapy and astragalus injection reduces blood viscosity, improves blood rheology and alleviates clinical svmptoms of blood stasis syndrome in senile cerebral infarction.