| Objective: Platelet activation and aggregation enhancement are the key factors of clinical thrombotic events.The increase of platelet parameters,especially MPV and PDW,is an important monitoring index of platelet activation and aggregation enhancement.Platelet activation and aggregation enhancement increases the risk of acute myocardial infarction and stroke,while platelets show obvious high reactivity in patients with diabetes,which also increases the incidence of cardiovascular disease in patients with diabet es.Gremlin-1 is a secretory glycosylated protein.The increase of plasma concentration can increase the occurrence of myocardial infarction.Recently,it has been found that Gremlin-1 is stored in resting platelets,which is released and enters circulation when platelets are activated.Increased platelet reactivity in diabetic patients can increase the incidence of cardiovascular disease,and the increase of plasma concentration of Gremlin-1 can also increase the incidence of myocardial infarction,so we speculate whether Gremlin-1 can enhance platelet activation and aggregation in diabetic patients and affect the occurrence of cardiovascular disease.The purpose of this study is to study the correlation between Gremlin-1,MPV and PDW through clinical experiments,and to study platelet activation and aggregation after intervention with Gremlin-1 reagent and IP3 blocker in vitro,so as to provide a theoretical basis for clinical prevention of thrombotic complications in patients with diabetes.Method: A total of 63 patients with diagnosed type 2 diabetes who were hospitalized in the cardiovascular department of our hospital from October 2019 to October 2020 were selected as the experimental group,and 20 healthy volunteers in the same period were selected as t he normal control group.The supernatant of 5m L venous blood was extracted after centrifugation,and the concentration of Gremlin-1 in serum was detected by ELISA method to observe the difference of expression between the two groups.In vitro,the whole bl ood of 10 healthy volunteers was collected and centrifuged with 10 m L,and the supernatant was taken to prepare platelet-rich plasma.After platelet activation was induced by ADP(10 μ M),Gremlin-1(20 μ g / m L)and ADP(10 μ M),platelet activation was induced by adding Gremlin-1(20μ g / m L)and ADP(10 μ M).After in vitro intervention with different concentrations of Gremlin-1(0 μ g / m L,10 μ g / m L,20 μ g / m L,30 μg / m L),platelet aggregation was detected by platelet aggregation meter to evaluate platelet aggregation.At the same time,ELISA method was used to detect the concentration of PF4 after intervention and to evaluate platelet activation.After admission,all the selected researchers completed glycosylated hemoglobin(Hb A1c),total cholesterol(TC),triglyceride(TG),high density lipoprotein(HDL-C),low density lipoprotein(LDL-C),white blood cell count(WBC),serum creatinine(Cr),C-reactive protein(CRP),platelet count(PLT),large platelet ratio(PLCR),platelet distribution width(PDW),(MPV)index of mean platelet volume was detected.Result:(1)The plasma Gremlin-1 concentration in the diabetic group was significantly higher than that in the control group(272.69±24.76 vs.223.11 ±22.46,P < 0.05).(2)The correlation analysis between plasma Gremlin-1 and MPV,PDW,Hb Alc,CRP showed that there was a significant positive correlation between Gremlin-1 and MPV,between Gremlin-1 and PDW,between Gremlin-1 and Hb Alc,and between Gremlin-1 and CRP(R = 0.376,P < 0.05,0.376,P < 0.05).The results showed that there was a significant positive correlation between Gremlin-1 and CRP(R= 0.278,P < 0.05),between Gremlin-1and Hb Alc(R = 0.289,P < 0.05),and between Gremlin-1 and CRP(R= 0.376,P < 0.05).(3)The effect of Gremli n-1 on platelet activation and aggregation was studied.The results showed that the level of PF4 in Gremlin-1+ADP group(19.99 ±3.19 ng/m L)and ADP group(16.91±3.05ng/m L)was significantly higher than that in Gremlin-1 group(12.67 ±2.37 ng/m L)and control group(11.52 ±2.16 ng/m L).There was significant difference between these two groups.Gremlin-1 group was also higher than control group,but there was no statistical difference.The platelet aggregation rate in Gremlin-1+ADP group(41.70 ±2.23%)and ADP group(24.36±3.64%)was significantly higher than that in Gremlin-1 group(11.98 ±2.01%)and control group(11.06 ±1.36%).The platelet aggregation rate in Gremlin-1 group was also higher than that in control group,but there was no statistical differe nce.(4)To study the effects of different concentrations of Gremlin-1 on platelet activation and aggregation induced by ADP.The results showed that the platelet aggregation rates induced by ADP(10 μ m)were 0 μ g / m L,10 μ g / m L,20 μ g / m L,30 μ g / m L,0 μ g / m L(23.50 ±2.23%),10 μg / m L(30.50 ±2.23%)and 20 μ g / m L(41.70 ±2.23%).The platelet aggregation rate of 30 μ g / m L group(42.30 ±2.23%)increased with the increase of Gremlin-1 concentration.The platelet aggregation rate of 0 μ g / m L group was significantly higher than that of the other three groups.The platelet aggregation rate of 10 μ g / m L group was significantly higher than that of 20 μ g / m L group and 30 μ g / m L group.The platelet aggregation rate of 20 μ g / m L group was higher than that of 30 μ g / m L group.But there was no significant difference(P > 0.05).The effects of Gremlin-1 on platelet activation and aggregation induced by ADP in different incubation time were as follows: the platelet aggregation rate induced by A DP(10 μ M)was incubated with Gremlin-1(20 μ g / m L)for 0 min,15 min,30 min and45 min,respectively.0 min group(14.20 ±2.07%),15 min group(31.90±2.07%),30 min group(41.70 ±2.07%),0 min group(14.20 ±2.07%),15 min group(31.90 ±2.07%),30 min group(41.70 ±2.07%).The platelet aggregation rate in the 45-minute group(42.90 ±2.07%)increased with the increase of incubation time,and the platelet aggregation rate in the 0-minute group was significantly higher than that in the other three groups(P < 0.05).The platelet aggregation rate in the 15-minute group was significantly higher than that in the30-minute group and 45-minute group(P < 0.05).The platelet aggregation rate in the 30-minute group was significantly higher than that in the 45-minute group(P < 0.05).But there was no significant difference(P > 0.05).Conclusion: 1.The level of Gremlin-1 in plasma of patients with diabetes is significantly increased,and the level of Gremlin-1 is positively correlated with platelet morphological parameters MPV and PDW.2.Gremlin-1 can enhance platelet activation and aggregation induced by ADP. |
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