| Objective:To explore possible mechanism of protective effects of estrogen on clopidogrel-induced gastric mucosal damage.Methods:Firstly,the relevant clinical data of hospitalized patients with gastric ulcer in the Zhongda Hospital affiliated to Southeast University were collected.Patients without pathological data or pathological data were not supported.The patients were divided into two groups(with clopidogrel).One group of medical history,the other group was patients who did not take clopidogrel),and then the pathological wax samples of the two groups were taken for immunohistochemical staining to observe the expression of ERa,and the two groups of patients were observed by semi-quantitative method.Expression in the gastric mucosa.Step-by-step verification was performed by cell experiments and animal experiments.In cytological experiments,GES-1 cells were selected as experimental cells.MTT method was used to detect the changes of cell viability under clopidogrel and estrogen.Immunofluorescence method was used to determine the expression of estrogen-related receptors and ROS(reactive oxygen species)under the action of drugs.Flow cytometry was used to detect the changes of apoptotic rate of cells under the action of different drugs.Microplate method was used to detect changes in oxidative stress-related factors under different conditions.In the animal experiment,12-week SD rats(Sprague Dawley Rat)were used as experimental animals.The gastric ulcer model was established by surgery.After feeding for 25 days,the ulcers were self-healed and then divided the rats into three groups(control group,clopidogrel group,clopidogrel+ estrogen group)to verify the different effects of clopidogrel and estrogen on healed ulcers.After 30 days,the rats were sacrificed and the gastric mucosa was examined for Western blot(WB)detection of related proteins and receptors.Results:In the study of human pathological wax specimens,the expression level of ERa was significantly lower in patients taking clopidogrel than in patients not taking clopidogrel(14.47±4.75 Vs 8.58±2.11).Clopidogrel can increase the expression of ROS and inhibit the expression of GSH(glutathione)and SOD(Superoxide Dismutase)by aggravating the oxidative stress in cells to aggravate cell damage and apoptosis(Control,Clopidogrel,Estrogen group SOD :10111.3±94.89,6150±12.00,9032±128.67;GSH : 71.8±2.03,65.8±0.95,57.2±1.12,respectively),p<0.05.In case of clopidogrel,estrogen significantly reduced the rate of apoptosis(0.586±0.0009vs0.383±0.0001),p<0.05 and inhibited the process of oxidative stress.In the animal experiment,it can be seen that there is obvious recurrence of ulcer in the clopidogrel group,and there is still bleeding.In contrast,the estrogen group can show significant ulcer healing performance,and no obvious bleeding or perforation is observed.The results of WB showed that estrogen-related receptors such as ERa,ER36 and GPR30 were significantly changed in the clopidogrel group,and ER? remained basically stable in the three groups.At the same time,the apoptotic protein and oxidative stress-related protein Nrf2 also increased significantly in the clopidogrel group,suggesting that there is aggravation of apoptosis.The proportion of PMEKK3/MEKK3 was significantly higher in the clopidogrel group than in the other two groups,suggesting that clopidogrel may play a role in promoting apoptosis through the MAPK/MEKK3/p38 pathway.Conclusion:Clopidogrel can promote the apoptosis of gastric mucosa and cells.Estrogen has obvious inhibitory effect on this effect and has protective effect on gastric mucosa.It may play a role in attenuating oxidative stress and reducing apoptosis,accompanied by activating the MKK3/p38 pathway.Further studies are still needed to. |
PDF Full Text Request