Effect of Xuebijing injection on endothelial microparticles and renal cortical microcirculation in septic rats / 中华危重病急救医学

Objective:To clarify the characteristics of renal cortical microcirculation and its relationship with the expression of plasma endothelial microparticle (EMP) in septic rats, and to evaluate the effect of Xuebijing injection as an adjuvant therapy of antibiotics on septic AKI.Methods:The 8-10 weeks old specific pathogen free (SPF) male Sprague-Dawley (SD) rats were divided into sham operation group (Sham group), positive drug control group and Xuebijing group by the random number table method, with 10 rats in each group. The cecal ligation and puncture (CLP) with large ligation (ligated 75% of the cecum) was used to prepare a rat high-grade sepsis model; in the Sham group, the cecum was stretched without ligation or puncture. Due to the high mortality of CLP with large ligation, Xuebijing injection (4 mL/kg, 12 hours per time) and imipenem/cilastatin injection (90 mg/kg, 6 hours per time) were administered to the rats in the Xuebijing group via the tail vein immediately after the model was produced. Normal saline and imipenem/cilastatin were administered to the rats by the same methods in the positive drug control group. The rats in the Sham group were treated with the same volume of normal saline as any of the other two groups at the same frequency. At 48 hours after model reproduction, the mean arterial pressure (MAP) and blood lactic acid (Lac) of the rats were measured. The renal cortical microcirculation was monitored by using side stream dark-field imaging. Renal hypoxia signals were assessed by pimonidazole chloride immunohistochemistry. Plasma EMP levels were determined by using flow cytometry, and then the correlation between EMP and microcirculation parameters of renal cortex was analyzed. At the same time, the serum creatinine (SCr) was measured, and the renal injury score (Paller score) was used to evaluate the severity of renal tissue pathological damage.Results:Compared with the Sham group, perfused vessel density (PVD), microvascular flow index (MFI) and MAP in the positive drug control group and the Xuebijing group decreased significantly, the positive expression of hypoxia probe (pimonidazole) increased, Lac, EMP, Paller score and SCr increased significantly. However, compared with the positive drug control group, the renal cortical microcirculation in the Xuebijing group was improved significantly, PVD and MFI were increased significantly [PVD (mm/mm 2): 16.20±1.20 vs. 9.77±1.12, MFI: 2.46±0.05 vs. 1.85±0.15, both P < 0.05], Lac was reduced significantly (mmol/L: 4.81±1.23 vs. 6.08±1.09, P < 0.05), MAP increased slightly [mmHg (1 mmHg = 0.133 kPa): 84.00±2.00 vs. 80.00±2.00, P > 0.05], suggested that Xuebijing injection improved renal microcirculation perfusion in septic rats, and this effect did not depend on the change of MAP. The positive expression of pemonidazole in renal cortex of the Xuebijing group was significantly lower than that of the positive drug control group [(35.89±1.13)% vs. (44.93±1.37) %, P < 0.05], suggested that Xuebijing injection alleviated renal hypoxia. The plasma EMP levels of rats in the Xuebijing group were significantly lower than those in the positive drug control group (×10 6/L: 3.49±0.17 vs. 5.78±0.22, P < 0.05), and the EMP levels were significantly negatively correlated with PVD and MFI ( r values were -0.94 and -0.95, respectively, both P < 0.05), indicated that the increase of plasma EMP was highly correlated with renal microcirculation disorder, and Xuebijing injection inhibited the increase of plasma EMP levels. The Paller score in the Xuebijing group was significantly lower than that in the positive drug control group (46.90±3.84 vs. 62.70±3.05, P < 0.05), and the level of SCr was also significantly lower than that in the positive drug control group (μmol/L: 121.1±12.4 vs. 192.7±23.9, P < 0.05), which suggested that Xuebijing injection relieved kidney injury and improved renal function in septic rats. Conclusion:As an adjuvant therapy of antibiotics, Xuebijing injection could inhibit the expression of plasma EMP in rats with sepsis, improve renal cortex microcirculation, and reduce kidney injury.

Effect of Endothelial Microparticles Induced by Hypoxia on Migration and Angiogenesis of Human Umbilical Vein Endothelial Cells by Delivering MicroRNA-19b / 中华医学杂志(英文版)

Background@#Microparticles (MPs) are small extracellular plasma membrane particles shed by activated and apoptotic cells, which are involved in the development of atherosclerosis. Our previous study found that microRNA (miR)-19b encapsulated within endothelial MPs (EMPs) may contribute to the upregulation of circulating miR-19b in unstable angina patients. Hypoxia is involved in atherosclerosis as a critical pathological stimulus. However, it still remains unclear whether the increase of miR-19b levels in EMPs is related to hypoxia and if the effect of miR-19b - wrapped within EMPs - stimulates hypoxia on vascular endothelial cells. This study aimed to explore the changes of miR-19b in EMPs induced by hypoxia as well as their effects on endothelial cells.@*Methods@#Human umbilical vein endothelial cells (HUVECs) were cultured in vitro and arranged to harvest EMPs in two parts: the first part consisted of EMP and EMP and the second part included EMP, EMP, and EMP. Cell migration was detected by scratch migration and transwell chamber migration. Angiogenesis was assessed by tube formation assays. Furthermore, we predicted the target gene of miR-19b by bioinformatics analysis, and luciferase assay was used to verify the targeted gene of miR-19b. Data were analyzed by one-way analysis of variance. Student's t-test was used when two groups were compared.@*Results@#Compared with EMP- and EMP-inhibited migration of cells by scratch migration assay (80.77 ± 1.10 vs. 28.37 ± 1.40, P < 0. 001) and transwell chamber migration assay (83.00 ± 3.46 vs. 235.00 ± 16.52, P < 0.01), the number of tube formations was markedly reduced by 70% in the EMP group (P < 0.001) in vitro analysis of HUVECs. Meanwhile, a strong inhibition of migration and tube formation of HUVECs in the presence of miR-19b-enriched EMP was observed. This effect might be due to the delivery of miR-19b in EMPs. Transforming growth factor-β2 (TGFβ2) was predicted to be one of the target genes of miR-19b, and we further confirmed that TGFβ2 was a direct target gene of miR-19b using the luciferase assay. The expression of TGFβ2 in HUVECs was inhibited by treatment with EMP and EMP.@*Conclusions@#MiR-19b in EMPs induced by hypoxia could reduce endothelial cell migration and angiogenesis by downregulating TGFβ2 expression, which may have inhibited the progression of atherosclerosis.

Correlation of endothelial inflammation in patients with acute myocardial infarction with no-reflow and severity of coronary lesion / 西安交通大学学报(医学版)

Objective To investigate the correlation of endothelial inflammation in patients with acute myocardial infarction (AMI)with no-reflow and severity of coronary lesion.Methods A total of 120 patients with AMI were all treated with percutaneous coronary intervention (PCI).The patients who had intraoperative no-reflow were in case group (n=40).The patients who were also treated by PCI but did not have no-reflow in the same period in our hospital served as the control group (n=80).According to the results of coronary angiography, no-reflow group patients were divided into three subgroups:single lesion group (n=14),two-lesion group (n=10), and three-lesion group (n=1 6 ).We monitored these patients’immediate postoperative endothelial microparticles (EMPs)and soluble intercellular adhesion molecule-1 (sICAM-1 )levels in both groups with flow cytometry and ELISA methods.Results In the case group,immediate postoperative EMPs and sICAM-1 levels in the blood circulation were significantly higher than those in the control group.EMPs area and sICAM-1 area under the curve in the case group were 0.755 and 0.701,which showed that EMPs and sICAM-1 levels had a good correlation with the occurrence of non-reflow in AMI patients after PCI.Gensini score and EMPs level rose with the increase of the lesion counts,and the level of EMPs was positively correlated with Gensini score.Conclusion In patients with AMI,the EMPs and sICAM-1 levels can well predict the occurrence of no-reflow and are of some value in predicting the severity of coronary lesion.

Changes of Plasma Endothelial Microparticles in Elder Patients With Acute Coronary Syndrome Combining Diabetic Mellitus / 中国循环杂志

Objective: To explore the endothelial microparticle (EMP) level in elderly patients with acute coronary syndrome (ACS) combining diabetic mellitus (DM)) and to study the relationship between EMP level and ACS combining DM in elder patients. Methods: A total of 208 patients≥65 years with coronary angiography in our hospital were summarized including 124 male with the age of (71.9 ± 5.2) years. The patients were divided into 3 groups, Control group, n=51 normal subjects, ACS without DM group, n=83 and ACS+DM group, n=74. Plasma EMP levels were measured by FACSCalibur lfow cytometry as CD31+/CD42b-EMPS and the vascular stenosis degree was quantitatively calculated with Gensini score. Results: The CD31+/CD42b-EMPs level in ACS + DM group >ACS without DM group > Control group, all P Conclusion: Plasma CD31+/CD42b-EMPs level increased in elderly ACS patients and the elevation level related to vascular lesion degree/combining with DM, which indicated the endothelial dysfunction in such patients.

Changes of endothelial microparticles in children with Kawasaki disease and its relation to coronary artery lesions / 中国小儿急救医学

Objective To study the changes of plasma endothelial microparticles (EMP) in children with Kawasaki disease (KD) and its relation to coronary artery lesions (CAL).Methods The participants in this study were 30 children with KD (24 children with typical KD and 6 cases with incomplete KD).All KD patients met the diagnostic criteria established by the Japanese Kawasaki Disease Research Committee.According to the course of KD,3 phases were divided:the acute phase,the subacute phase and the convalescent phase.We evaluated the presence of CAL using two-dimensional echocardiographic examination,and then the KD children were divided into two groups,including 24 children without CAL and 6 children with CAL.Ten children with fever and rash and 10 healthy children were studied as control.The levels of CD31+/CD42b- EMP were measured by flow cytometry.Results The level of EMP was significantly higher in the acute phase [ (8.18 ± 2.29) ％ ] than those either in the convalescent phase [ (2.77 ± 0.85 ) ％ ] of KD or the healthy children [ ( 1.34 ± 0.38 ) ％ ] (P ＜ 0.01 ).The level of EMP was also significantly higher in the subacute phase [ (5.93 ± 1.05 )％ ] than those either in the convalescent phase of KD or the healthy children (P ＜0.01 ).The level of EMP was higher in the children with fever [ (3.66 ± 1.16) ％ ] than that in the healthy children ( P ＜ 0.05 ).Furthermore,the level of EMP during the acute phase was also higher in KD patients with CAL than in those without CAL(P ＜0.01 ).Conclusion The measurement of EMP may be useful for the early diagnosis of KD and the identification of CAL.

High glucose induces release of endothelial microparticles in human umbilical vein endothelial cells and promotes cell apoptosis / 西安交通大学学报(医学版)

Objective To explore the effects of high glucose on release of endothelial microparticles (EMPs) and cell apoptosis in human umbilical vein endothelial cells (HUVECs). Methods HUVECs were incubated for 0h, 12h, 24h, 48h and 72h, respectively, with different concentrations of glucose (5.5mmol/L, 16.7mmol/L and 33.3mmol/L, respectively). The cells cultured in the media containing glucose 5.5mmol/L and mannitol 27.8mmol/L served as control groups. The survival rate of cells was examined by MTT assay. The level of EMPs release and percentage of cell apoptosis were detected by flow cytometry. Results ① According to MTT detection, the survival rate of HUVECs was 93% when incubated for 12h with 33.3mmol/L, 78% for 24h, 51% for 48h, and 40% for 72h. Compared with that for 0h, the survival rate for 12h did not decline significantly (P>0.05). the survival rate reduced significantly for 24, 48 and 72h, respectively, in the time-dependent manner (P<0.05). ② If the survival rate was 100% when incubated in 5.5mmol/L glucose for 48h, then it was 98% in mannitol 27.8mmol/L with no significant difference (P>0.05). It was 79% in 16.7mmol/L glucose and 51% in 33.3mmol/L glucose, obviously lower than that in 5.5mmol/L glucose, and the two groups differed significantly (P<0.05). ③ the level of EMPs release increased significantly in 33.3mmol/L glucose for 12, 24, 48 and 72h respectively, in the time-dependent manner. Compared with the normal glucose group, the level of EMPs release did not significantly differ in mannitol 27.8mmol/L for 12, 24, 48 and 72h , respectively (P>0.05). ④ There was a significantly positive correlation between EMPs release and cell apoptosis (r=0.659, P<0.05). Conclusion High glucose induces release of HUVECs endothelial miccroparticles and promotes cell apoptosis.