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The injury of vascular endothelial cells is not only the initial condition to promote the occurrence of early atherosclerosis （AS） plaques， but also an important link in the pathogenesis of AS. The microRNA （miRNA）， as an important medium of intercellular communication and gene regulatory factor， can affect vascular endothelial function and participate in the development of AS. The molecular mechanism of miRNA’s multi-target intervention in vascular endothelial cell injury has become a hot topic in the research of cardiovascular diseases. Monomers of traditional Chinese medicines such as ginsenoside Rb2 and paeonol， as well as traditional Chinese medicine for resolving phlegm and removing blood stasis could regulate miRNA to improve endothelial cell inflammation； astragaloside Ⅳ， dihydromyricetin and notoginsenoside could target miRNA and inhibit vascular endothelial oxidative stress； Danhong injection， Jianpi qutan and huayu prescription and paeonol could affect endothelial autophagy through miRNA； resveratrol， Bushen huoxue formula and Bushen tongmai formula could inhibit vascular endothelial aging by miRNA； dendrobine played an active role in regulating miRNA and improving endoplasmic reticulum stress. In the future， more in- depth research is needed on the effectiveness， mechanism of action， diagnosis and treatment plans， and safety of targeted regulation of miRNA for AS therapy by traditional Chinese medicine.
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Aim To investigate whether CTSB is involved in hypoxia-induced injury of cardiac microvascular endothelial cells.Methods A hypoxia-induced endothelial cell injury model was used.Cardiac microvascular endothelial cells were isolated from CTSB gene knockout mice.CTSB was overexpressed by adenovirus delivery system, and bafilomycin was used to block autophagy.ELISA was used to detect the release of inflammatory factors.Tunel staining was used to detect the number of cell apoptosis.caspase-3 kit was used to detect the activity of cell caspase-3.Cells were infected with LC3-GFP-mCherry double-labeled adenovirus todetect cell autophagy flow.Results CTSB gene knockout could significantly aggravate the inflammation and apoptosis of endothelial cells induced by hypoxia, and increased autophagy.Overexpression of CTSB reduced the inflammation and apoptosis of endothelial cells induced by hypoxia, and increased autophagy.But bafilomycin treatment could significantly offset the inhibitory effect of CTSB overexpression on cell inflammation and apoptosis and the protective effect on cell autophagy.Conclusions CTSB knockout aggravates inflammation and apoptosis induced by hypoxia in endothelial cells; while the overexpression of CTSB ameliorates endothelial cell injury induced by hypoxia.CTSB maintains normal autophagy degradation in endothelial cells.BAF blocks the protective effect of CTSB on endothelial cells by inhibiting autophagy degradation.
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Diabetic nephropathy (DN) is one of the diabetic microvascular complications characterized by progressive protein uria and renal failure, which may eventually develop into end-stage renal disease (ESRD). Glomerular endothelial cells are one of the important components of glomerular filtration barrier. The structural and functional integrity of these cells are closely related to the maintenance of glomerular filtration function. Dysfunction of glomerular endothelial cells can lead to proteinuria, glomerulosclerosis and interstitial fibrosis, further damaging renal function and accelerating the progression of DN. The mechanisms leading to endothelial dysfunction include glucose metabolism disorder, oxidative stress, abnormal angiogenesis, inflammation and endothelial transdifferentiation. In recent years, it has been proposed that mechanisms such as intercellular communication, epigenetics and exosomes may be involved in the injury of diabetic nephropathy. In present paper, the research progress of related mechanisms was reviewed on the damage of glomerular endothelial cells in DN.
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Objective Find abnormal changes of plasma lipid metabolism-related proteins before 20 weeks of gestation in patients with hypertensive disorder of pregnancy(HDP), and preliminarily investigate the role of plasma apolipoprotein C4 elevation in HDP. Methods A nested case-control study was used. The plasma were collected from pregnant women who underwent routine prenatal examination in Guangzhou Women and Children's Medical Center from November 2014 to March 2017. Label-free mass spectrometry was used to detect the differences in plasma lipid metabolism-related proteins before 20 weeks of gestation between 12 pairs of HDP patients and normal controls, and different 48 pairs of samples were used for verification. The protein with the most significant difference multiples was screened to study its effects on monolayer permeability and nitric oxide secretion of endothelial cells. One-way ANOVA was used for comparison between groups, and P<0.05 was considered as statistically significant difference. Results Compared with the control, the lipid metabolism-related proteins, APOC4, Fatty acid-binding protein 4 (FABP4), Apolipoprotein E (APOE), Apolipoprotein C3 (APOC3) and Beta-2-glycoprotein 1(APOH) raised to 1.94, 1.82, 1.59, 1.55 and 1.38 times, phospholipid transfer protein (PLTP) decreased to 0.78 times in plasma before 20 weeks of pregnancy of patients with HDP (t value were 2.499, 2.497, 2.081, 2.098, 2.426 and 2.564, respectively, P<0.05). Cell experiments results showed that 50 ng / ml APOC4 significantly increased 20％ HUVEC single layer cell permeability to FITC-labeled dextran (F=455.4, P<0.01), and significantly decreased the level of nitric oxide in the supernatant of HUVEC culture by 25％ (F=61.92, P<0.01). Conclusions Before diagnosis, plasma protein levels involved in lipid metabolism in HDP patients have been changed, resulting in abnormal lipid metabolism. APOC4 can increase the permeability of vascular endothelial cells, inhibit endothelial source of NO secretion, cause endothelial dysfunction.
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Objective To investigate the protective function of endovascular cooling method on post-resuscitation syndrome (PRS) in porcine cardiac arrest (CA) model and its mechanism.Methods Ventricular fibrillation (VF) was electrically induced and untreated for 8 minutes in 15 healthy male porcines, cardiopulmonary resuscitation (CPR) was then initiated. All successful recovery animals were randomly divided into two groups by random number table. In normal temperature group, the core temperature was maintained at (38.0±0.5) ℃ for 12 hours. In mild hypothermia group, the mild hypothermia treatment was initiated at 5 minutes after successful resuscitation, the treatment of rapid endovascular cooling was performed to reach the target cooling temperature of (33.0±1.0) ℃, and then maintained until 6 hours after resuscitation. Rewarming was implemented at the rate of 0.7 ℃/h until the body temperature reached (38.0±0.5) ℃. Hemodynamic parameters including heart rate (HR), mean arterial blood pressure (MAP), cardiac output (CO) were continually monitored. Right femoral vein blood was collected before VF and 1, 2, 4, 6, 12 and 24 hours after resuscitation, respectively, and the serum concentrations of E-selectin, soluble thrombomodulin (sTM), and interleukin-1β(IL-1β) were determined with enzyme linked immunosorbent assay (ELISA). The survival of porcines at 24 hours after resuscitation was observed, and the neurological deficit score (NDS) was calculated for the surviving porcines. All animals were sacrificed, and brain, heart and lung tissues were collected, after hematoxylin and eosin (HE) staining, the histopathology changes were evaluated under a light microscopy.Results After 8-minute VF, 14 porcines were resuscitated successfully, 7 porcines in normal temperature group and 7 in mild hypothermia group respectively, with the resuscitation success rate of 93.3％. There was no significant difference in body weigh, core temperature, hemodynamics, or blood lactate as well as duration of CPR and the number of defibrillations between the two groups. The core temperature of normal temperature group was maintained at (38.0±0.5) ℃, while in mild hypothermia group, the hypothermia was reduced to the hypothermia range (33.0±1.0) ℃until 6 hours, then rewarmed to normothermia gradually [(38.0±0.5) ℃]. Compared with those before VF, HR was significantly increased after resuscitation in both groups, and MAP and CO were decreased, then they tended to normal. There was no significant difference in hemodynamic parameter at all time points between the two groups. Compared with those before VF, the levels of E-selectin and sTM in serum of the two groups were increased significantly at 1 hour after resuscitation, and they were decreased gradually after reaching the peak at 6 hours, and IL-1β was increased continuously with time. There was no significant difference in E-selectin (μg/L:1.34±0.52 vs. 1.60±0.61), sTM (μg/L: 19.13±0.34 vs. 19.24±0.73), or IL-1β (ng/L: 25.73±0.87 vs. 25.32±0.25) before VF between normal temperature group and mild hypothermia group (allP> 0.05). The levels of E-selection, sTM and IL-1β in mild hypothermia group were significantly lower than those in normal temperature group from 2 hours after resuscitation [E-selection (μg/L): 11.15±2.73 vs. 16.04±3.23, sTM (μg/L): 49.67±3.32 vs. 62.22±1.85, IL-1β (ng/L): 140.51±6.66 vs. 176.29±18.51, allP< 0.05], and E-selection decreased to the baseline level at 12 hours (μg/L: 1.17±0.65 vs. 1.60±0.61,P > 0.05). The 24-hour survival rates of two groups were both 100％. The NDS score of mild hypothermia group was obviously lower than that of normal temperature group (150.0±6.6 vs. 326.4±12.3,P < 0.05). In normal temperature group, neuronal cell necrosis was observed in the cerebral cortex at 24 hours after resuscitation, and nucleus was deeply stained. The myocardial necrosis and alveolar collapse was found. Meanwhile the infiltration of inflammatory cell could be found in the myocardium and alveolar. The brain, lung and myocardium injury were significantly milder in mild hypothermia group as compared with those in normal temperature group.Conclusions The intravascular cooling therapy was a safe and effective method for inducing mild hypothermia after resuscitation. This cooling effect was fast and reliable, and the rewarming speed was controllable and stable. The protective mechanism of mild hypothermia on PRS may be related to inhibiting systemic inflammatory response and reducing vascular endothelial cell injury.
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Objective To explore the safety and therapeutic effect of Xuebijing injection for treatment of patients with capillary leak syndrome (CLS).Methods Seventy-seven patients with clinical diagnosis of CLS admitted to Intensive Care Unit (ICU) of the First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine (TCM) from November 2015 to October 2016 were enrolled, they were divided into a control group (35 cases) and a Xuebijing group (42 cases) according to random number table method. The conventional treatment was given and at the same time the primary disease was actively treated in the control group; while in the Xuebijing group, on the basic treatment of the control group, additionally, Xuebijing injection 100 mL+ 0.9% normal saline (100 mL) was intravenously dripped, twice a day, 5 days constituting one therapeutic course. Before and after treatment for 5 days, the white blood cell count (WBC), neutrophils percentage (N), alanine transaminase (ALT), aspartate transaminase (AST), blood urea nitrogen (BUN), serum creatinine (SCr), procalcitonin (PCT), pH value, partial pressure of blood oxygen (PaO2), blood lactic acid value (Lac), activated partial thromboplastin time (APTT), prothrombin time (PT), blood platelet count (PLT) in the patients of the two groups were compared; and the acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score was recorded; the length of stay in ICU, mechanical ventilation time, and 28-day survival rate were statistically calculated in two groups.Results After treatment, the levels of WBC, N, PCT, ALT, AST, BUN, SCr, Lac, APACHE Ⅱ score in Xuebijing group were lower than those in the control group [WBC(×109/L): 9.85±0.61 vs. 13.87±2.58, N: 0.75±0.08 vs. 0.90±0.10, PCT (μg/L): 1.13±0.71 vs. 4.99±1.38, ALT (U/L): 79.56±30.85 vs. 84.21±27.32, AST (U/L): 91.98±38.10 vs. 110.28±35.79, BUN (mmol/L): 7.35±0.82 vs. 8.57±1.43, SCr (μmol/L): 111.67±43.96 vs. 132.51±55.10, Lac (mmol/L): 1.88±1.01 vs. 3.31±1.46, APACHE Ⅱ score: 11.34±3.59 vs. 17.65±4.77]; the PaO2, PLT, 28-day survival rate in Xuebijing group were higher than those in the control group [PaO2 (mmHg, 1 mmHg = 0.133 kPa): 75.47±21.10 vs. 54.22±15.23, PLT (×109/L): 211.54±58.25 vs. 153.27±49.69, 28-day survival rate: 85.71% (36/42) vs. 71.43% (25/35), allP < 0.05]; the PT, APTT, ICU hospitalization time and mechanical ventilation time in Xuebijing group were shorter than those in the control group [PT (s): 13.62±2.11 vs. 18.45±4.26, APTT (s): 31.33±4.27 vs. 36.85±5.56, length of stay in ICU (days): 12.4±3.7 vs. 20.5±4.1, mechanical ventilation time (days): 10.5±4.9 vs. 18.7±5.5, allP < 0.05].Conclusion The application of Xuebijing injection for treatment of patients with CLS can relieve their disease situation, reduce inflammatory indicators, improve the blood coagulation function and hypoxemia, shorten the ICU hospitalization time and mechanical ventilation time, elevate the 28-day survival rate, and has no harmful effects on liver and kidney functions.
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AIM:To investigate the expression of visfatin in the placenta of patients with preeclampsia and its significance.METHODS:The pregnant women (n=100) were divided into normal pregnancy group , mild preeclampsia group and severe preeclampsia group according to the severity of the disease .The pathological changes of the placenta were observed by hematoxylin-eosin staining .The expression of visfatin at mRNA and protein levels in the placenta was detected by real-time PCR and immunohistochemistry respectively .RESULTS:Compared with normal pregnancy group , the patho-logical changes of the placenta in preeclampsia groups was significant , showing that the structure and the form were disor-dered and incompleted in both cytotrophoblasts and syncytiotrophoblasts .The proliferation of cytotrophoblasts and the num-bers of the placental villi with syncytial knots were observed .The vascular numbers of villi were decreased and congested . The results of immunohistochemistry and real-time PCR showed that the visfatin protein was observed in the cytoplasm of cy-totrophoblasts and syncytiotrophoblasts among 3 groups.The expression of visfatin at mRNA and protein levels was in-creased with the severity of the preeclampsia .CONCLUSION:The injury and dysfunction of vascular endothelial cells in the villi exist in the patients with preeclampsia .High expression of visfatin at mRNA and protein levels in placenta of the patients with preeclampsia indicates that visfatin is closely related to preeclampsia .
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This article was aimed to study the protective effects and mechanism of resveratrol (Res) on endothelial injury induced by atherosclerosis ( AS ) . Lysophosphatidylcholine ( LPC ) was applied to induce the model of injured endothelial cells. Flow cytometer was used to detect the rate of cell apoptosis. TUNEL staining was used to detect the cell apoptosis. MTT colorimetric test was used to detect the cell viability. Automatic biochemistry analyzer was used to measure the LDH level . ELISA was used to detect TNF-α level . RT-PCR was used to determine the mRNA levels of calcitonin gene-related peptide (CGRP). The results showed that Res with the concentration of 20 μmol/L can effectively slow down the degree of endothelial cell injury induced by LPC which caused the cell disability, high LDH level and high TNF-α level (P < 0.01). Res showed effects on mRNA expression of CGRP. It was concluded that Res can protect against endothelial injury induced by AS, the mechanism of which may be associated with accelerating CGRP synthesis and release by activating VR1.
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@#BACKGROUND: Current studies on CD62P have focused mainly on cardiovascular diseases, while only few studies have evaluated the effects of CD62P on the development of sepsis and the association between endothelial cell injury with inflammation and coagulation. This study attended to explore the association between endothelial cell injury with inflammation and coagulation by evaluating the expression of soluble CD62P (s-CD62P) in plasma and its mechanism in patients with sepsis, thus to provide the evidence of effective treatment of sepsis with anti-adhesion therapy targeted CD62P. METHODS: A total of 70 critically ill patients with systemic inflammatory response syndrome (SIRS) admitted to intensive care unit (ICU) between September 2009 and February 2010 were enrol ed for a prospective and control study. According to the diagnostic criteria of sepsis/SIRS, the patients were divided into two groups: a sepsis group (n=38) and a SIRS group (n=32). Another 20 healthy volunteers served as a control group. Patients in the sepsis group and SIRS group were matched by clinical signs of high blood pressure, diabetes and its complications. The demographics of the patients including age, sex, body mass index (BMI), smoking and alcohol addict were compared among the groups. Six mL peripheral blood samples were collected within 24-hour admission in ICU for enzyme-linked immunosorbent assay (ELISA) to detect the plasma levels of s-CD62P, TNF-α, and hs-CRP. And variables of coagulation function such as platelet (PLT), prothrombin (PT), activated partial thromboplastin time (APTT), D-dimer and antithrombin-III (AT-III) were analyzed during 24 hours after admission to ICU. Meanwhile sequential organ failure assessment (SOFA) score of critically ill patients was evaluated. Data were expressed as mean±standard deviation and were statistical y analyzed by using SPSS 17.0 statistical software. The differences in plasma levels of s-CD62P of patients in each group were analyzed by ANOVA and the Kruskal-Wallis test. The relations between s-CD62P and inflammatory cytokines as well as with coagulation were determined by Pearson's product moment correlation coefficient analysis. Changes were considered as statistically significant if P value was less than 0.05. RESULTS: Compared with the control group and SIRS group, the sepsis group demonstrated significantly higher levels of s-CD62P, TNF-α and highly sensitive C-reactive protein (hs-CRP) (P<0.05). The plasma levels of D-dimer, PT, and APTT in the sepsis and SIRS groups were significantly higher than those in the control group, while the platelet count and the activity of AT-III were obviously lower (P<0.05). In the sepsis group, the plasma levels of hs-CRP and TNF-α were positively correlated with PT, APTT, and D-dimer, and negatively correlated with AT-III and PLT (P<0.05). The plasma levels of s-CD62P were significantly correlated with the plasma levels of TNF-α, hs-CRP, D-dimer, PT, and APTT, whereas they were correlated negatively well with PLT and AT-III (P<0.05). CONCLUSIONS: The concentration of plasma s-CD62P is elevated as a early biomarker in patients with sepsis, and it serves as one of the pathogenic factors responsible for endothelial cell damage. Coagulation and mediators of inflammation promote each other, aggravating the severity of sepsis. Plasma s-CD62P may be an important factor for the development of coagulation and inflammatory reaction.
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Objective To observe the clinical findings about the endothelial cell injury related to the genesis of inflammatory cytokines and coagulation.Methods A total of 70 critically ill patients with SIRS (systemic inflammatory response syndrome) admitted to intensive care unit (ICU) between September 2009 and February 2010 were enrolled for a prospective and control study.According to diagnostic criteria of Sepsis/SIRS,the patients were divided into two groups:sepsis group (n =38) and SIRS group (n =32),and another 20 healthy volunteers served as control group.Patients in the sepsis group and SIRS group were matched by clinical signs of high blood pressure,diabetes and its complications.The demographics of patients including age,sex,body mass index (BMI),smoking and alcohol addict were comparable among the different groups.The 6 ml peripheral blood samples were collected within 24 h after admission to ICU for enzyme-linked immunosorbent assay (ELISA) to detect the plasma levels of s-CD62P,TNF-α,and hsCRP.And variables of coagulation function such as platelet (PLT),prothrombin time (PT),activated partial thromboplastin time (APTT),D-dimer and antithrombin-Ⅲ (AT-Ⅲ) were analyzed during 24 h after admission to ICU.Meanwhile sequential organ failure assessment (SOFA) score of critically ill patients was evaluated.Data were expressed in mean ± standard deviation and were statistically analyzed by using SPSS 17.0 statistical software.The differences in plasma levels of s-CD62P of patients in each group were analyzed by ANOVA and Kruskal Wallis test.The relationship between s-CD62P and inflammatory cytokines as well as with coagulation were determined by Pearson correlation analysis.Changes were considered as statistically significant if P value was less than 0.05.Results ① Compared with control group and SIRS group,the levels of s-CD62P,TNF-α and high sensitive C-reactive protein (hs-CRP) were significantly higher in sepsis group (P ＜ 0.05).② The plasma levels of D-dimer,PT,APTT in sepsis group and SIRS group were significantly higher than those in control group,while the platelet count (PLT) and the activity of AT-Ⅲ were obviously lower (P ＜ 0.05).③ In sepsis group,the plasma levels of hs-CRP and TNF-α positively correlated with PT,APTT,D-dimer,and negatively correlated with AT-Ⅲ,PLT (P ＜ 0.05).④ Plasma levels of s-CD62P were significantly correlated with plasma levels of TNF-α,hs-CRP,D-dimer,PT,APTT,whereas correlated negatively well with PLT,AT-Ⅲ (P ＜ 0.05).Conclusions The plasma s-CD62P concentration is elevated as a early biomarker in patients with sepsis,and it acted as one of pathogenic factors responsible for endothelial cell damage.Coagulation and mediators of inflammation promotes each other,aggravating the severity of the sepsis.The plasma s-CD62P may be the important factor associated with initiation of coagulation development and inflammatory reaction.
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Objective To investigate the pulmonary microvascular responsiveness of diabetic animals to sepsis and the potential mechanism of NO system.Methods Sixty-four Wistar rats of clean grade were randomly (random number) divided into 4 groups,namely normal control group (group A,n =16),diabetes group (group B,n =16),sepsis group (group C,n =16),diabetes and sepsis group (group D,n =16).Diabetic mellitus model was made in rats with injection of streptozotocin,STZ (65 mg/kg).Successful model was defined as the blood glucose value≥ 16.67 mmol/L 48 hours after injection of STZ.All animals were fed 4 weeks before initiation of next experiment.The sepsis model was established by intravenous injection of LPS (10 mg/kg) in rats.RT-PCR was used to determine the mRNA expression of Tie-2 in rats'blood.The ratio of dry/wet of lung tissue and the extravasation of Evans blue dye into the lung were detected.Quantitation of NO in lung tissue and serum was measured by using Griess method.RT-PCR was also used for determination of iNOS,eNOS,DDAH2 mRNA expressions in lung tissue.Data were analyzed with ANONA and LSD method for comparison between groups,and P ＜ 0.05 was considered statistically significant.Results Compared with septic group.,the diabetic rats with sepsis group demonstrated higher expression of Tie-2 mRNA in blood (19.72 ± 0.70) vs.(3.99 ± 0.92),P =0.00,lower ratio of dry/wet in lung tissue (0.19 ±0.01) vs.(0.22 ±0.01),P =0.000,higher permeability of Evans blue dye into lung tissue (3.76 ± 0.77) vs.(1.74 ± 0.24),P =0.000.Serum NO level was lower in group D than that in group C (123.13 ±4.24) vs.(188.30 ±5.18),P =0.000,however,NO levels in lung tissue of both group D and group C were higher than that in control group (53.62 ± 6.70),(23.63± 3.92) vs.(10.37 ± 1.29),P =0.00,and NO level in group D was higher in 2 times than that in group C (P =0.00).However,there were no differences in eNOS expression among groups A,B and C,but the difference in eNOS expression was present between group D with lower expression and group A,that lower in group D (0.07 ±0.02) vs.(0.38 ±0.05),P=0.017.Compared with group C,the expression of iNOS was higher in group D (80.23 ±2.49),(32.48±5.37) vs.(1.74±0.23),P=0.00),and the expression of DDAH2 was lower in group D (0.49 ±0.13),(7.26 ±0.50) vs.(11.96 ±0.55).Conclusions Diabetic rats with sepsis enhanced endothelial cell damages.Diabetes deteriorates the regulatory activity of NO system,suggesting the potential mechanism of the worsened damages of EC in diabetic sepsis host.