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Force-dependent calcium signaling and its pathway of human neutrophils on p-selectin in flow

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protein & Cell 2017年第2期8卷 103-113页

作者： Bing Huang Yingchen Ling Jiangguo Lin Xin Du Ying Fang Jianhua Wu School of Bioscience & Bioengineering South China University of Technology Guangzhou 510006 China Hematology and Oncology Division Guangdong General Hospital Guangzhou 510080 China

p-selectin engagement of p-selectin glycoprotein Iigand-1 （pSGL-1） causes circulating leukocytes to roll on and adhere to the vascular surface, and mediates intracellular calcium flux, a key but unclear event for subsequent arresting firmly at and migrating into the infection or injured tissue. Using a parallel plate flow chamber technique and intracellular calcium ion detector （Fluo-4 AM）, the intracellular calcium flux of firmly adhered neutrophils on immobilized p-selectin in the absence of chemokines at various wall shear stresses was investigated here in real time by fluorescence microscopy. The results demon- strated that p-selectin engagement of pSGL-1 induced the intracellular calcium flux of firmly adhered neutrophils in flow, increasing p-selectin concentration enhanced cellu- lar calcium signaling, and, force triggered, enhanced and quickened the cytoplasmic calcium bursting of neu- trophils on immobilized p-selectin. This p-selectin-induced calcium signaling should come from intracellular calcium release rather than extracellular calcium influx, and be along the mechano-chemical signal pathway involving the cytoskeleton, moesin and Spleen tyrosine kinase （Syk）. These results provide a novel insight into the mechano-chemical regulation mechanism for p-selectininduced calcium signaling of neutrophils in flow.

关键词： neutrophils p-selectin calcium signaling shear stress

Effect of platelet-derived p-selectin on Neutrophil Recruitment in a Mouse Model of Sepsis-induced Acute Kidney Injury

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Chinese Medical Journal 2017年第14期130卷 1694-1699页

作者： Xiu-Hua Li Yong-Bing Qian Xiao-Xiao Meng Rui-Lan Wang Department of Critical Care Medicine Shanghai General Hospital of Nanjing Medical University Shanghai 201620 China Emergency Center Shanghai East Hospital Tongji University School of Medicine Shanghai 200120 China

Background：Acute kidney injury （AKI） is a severe disease in critically ill patients.Neutrophil infiltration into kidney was associated with the development of AKI,and p-selectin may be involved in the process of neutrophil recruitment in kidney.This study aimed to explore the potential effect of platelet-derived p-selectin on neutrophil recruitment in a mouse model of sepsis-induced AKI.Methods：A total of 30 C57BL/6 male mice were divided into five groups （n =6 in each）：sham group,sepsis group,anti-Ly6G group,anti-p-selectin group,and platelet depletion group.Sepsis was induced by cecal ligation and puncture.Serum creatinine concentration and platelet activity were measured by biochemical detector and flow cytometry,respectively.Histological and pathological features were analyzed using hematoxylin-eosin （HE） and immunohistochemi stry （IHC） staining,respectively.Myeloperoxidase （M pO） activity was detected with MpO assay.Unpaired t-test was used for data analysis.Results：Serum creatinine increased significantly in septic group compared to sham group （2.68 ± 0.27 mg/dl vs.0.82 ± 0.19 mg/dl,t =12.06,p =0.0000） but attenuated in antibodies-treated animals compared to septic group （anti-Ly6G：1.62 ± 0.30 mg/dl vs.2.68 ± 0.27 mg/dl,t =5.76,p=0.0004;anti-p-selectin：1.76 ± 0.31 mg/dl vs.2.68 ± 0.27 mg/dl,t =4.92,p =0.0012;and platelet depletion：1.93 ± 0.29 mg/dl vs.2.68 ± 0.27 mg/dl,t =4.14,p =0.0032）.platelet amount significantly decreased compared to sham group （658.20 ± 60.64 × 109/L vs.822.00 ± 48.60 × 109/L,t =4.71,p =0.0015） in septic mice,especially in platelet depletion group （240.80 ± 44.98 × 109/L vs.822.00 ± 48.60 × 109/L,t =19.63,p =0.0000）.p-selectin activity was significantly increased in septic group compared to sham group （16.54 ± 1.60% vs.1.90 ± 0.29%,t =15.64,p =0.0000） but decreased significantly in platelet depletion group compared to septic group （3.62 ± 0.68% vs.16.54 ± 1.60%,t =12.89,p =0.0002）.IHC analysis shown that neutrophil infiltration increased in septic mice compared to sham group （36.67 ±

关键词： Acute Kidney Injury Neutrophils p-selectin Sepsis

Temporal changes in circulating p-selectin,plasminogen activator inhibitor-1,magnesium,and creatine kinase after percutaneous coronary intervention

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2010年第8期11卷 575-582页

作者： Shu-qin YING Mei-xiang XIANG Lu FANG Jian-an WANG Department of Cardiology Hangzhou 310009 China) Cardiovascular Key Lab of Zhejiang province the Second Affiliated Hospital School of Medicine Zhejiang University Hangzhou 310009 China Baker IDI Heart and Diabetes Institute Melbourne Victoria 3004 Australia

Objective:This study aims to determine the mechanisms underlying restenosis and ischemia-reperfusion injury of the myocardium after percutaneous coronary intervention(pCI).Methods:The present study examined serial changes(5 min,30 min,2 h,6 h,and 24 h after pCI) in circulating p-selectin,plasminogen activator inhibitor-1(pAI-1),magnesium(Mg),and creatine kinase-myocardial band fraction(CK-MB) levels,which may be associated with restenosis and myocardial injury in patients undergoing pCI.The occurrence rates of major adverse cardiovascular events were collected over a six-month follow-up.Results:pCI induced an early elevation of p-selectin,which correlated positively with the inflation pressure used in the pCI procedure.pCI also caused a significant and sustained decrease in serum Mg in pCI patients,without an effect on pAI-1.An increase in CK-MB was observed in pCI patients,although values were within normal reference range.In addition,elevated p-selectin and decreased Mg measured shortly after the coronary angioplasty procedure were associated with recurrent treatment and heart failure,respectively.Conclusions:Our study demonstrates that pCI induces temporal changes of p-selectin,Mg,and CK-MB,which may be involved in restenosis and ischemia-reperfusion injury.These findings highlight the need for using antiplatelet therapy and Mg to reduce the risks associated with pCI.

关键词：关键词经皮的冠的干预狭窄 Ischemia-reperfusion 损害 p-selectin 镁

Effect of Aqueous Extracts of Several Kinds of Herbs on Human platelet Aggregation and Expression of p-selectin in Vitro

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Chinese Journal of Integrative Medicine 2015年第4期21卷 286-290页

作者：严令耕阮君山张蕾范方田张峰王爱云郑仕中曾莉李文林陆茵 Department of Clinical pharmacy College of PharmacyNanjing University of Chinese Medicine Library of Nanjing University of Chinese Medicine Fujian provincial Hospital Clinical College of Fujian Medical University

Objective： To study the effect of aqueous extract of several kinds of herbs on human platelet aggregation and expression of p-selectin in vitro. Methods： Blood was collected from volunteers. Effects of the prepared water extracts of herbs on platelet aggregation were monitored on a packs-4 aggregometer. The fluorescence intensity of water extracts of Caulis Spatholobi, Flos Carthami and Rhizoma Curcumae on the expression of p-selectin in human platelets of healthy persons was measured with flow cytometry. Results： Out of several herbs investigated, Flos Carthami and Rhizoma Curcumae potently inhibited platelet aggregation after incubation with platelet-rich plasma （pRp） for 15 min. Caulis Spatholobi, Flos Carthami and Rhizoma Curcumae inhibited adenosine-5＇-diphosphate （ADp） or platelet activating factor （pAF）-induced platelet aggregation in pRp in a dose-dependent manner. In contrast to Flos Carthami and Rhizoma Curcumae, Caulis Spatholobi could not inhibit thrombin-induced platelet aggregation. Despite its inability to inhibit thrombin-induced platelet aggregation in pRp, Caulis Spatholobi had a greater anti-aggregating activity in pRp induced by ADp or pAF. Caulis Spatholobi and Flos Carthami showed significant inhibitory effects on the expression of p-selectin. Conclusions： Caulis Spatholobi, Flos Carthami and Rhizoma Curcumae have potent anti-platelet properties, and their inhibitory actions are mediated via different mechanisms. Caulis Spatholobi inhibited ADp-induced platelet aggregation but not by thrombin, indicating that its mechanism of action might be independent of the thromboxane pathway. The effect of Caulis Spatholobi and Flos Carthami were associated with suppressing the expression of p-selectin.

关键词： platelets herbs platelet aggregation Caulis Spatholobi Flos Carthami Rhizoma Curcumae,platelet-rich plasma p-selectin

Role of p-selectin and anti-p-selectin monoclonal antibody in apoptosis during hepatic/renal ischemia-reperfusion injury

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World Journal of Gastroenterology 2000年第2期6卷 244-247页

作者： pei Wu Xiao Li Tong Zhou Ming Jun Zhang Jin Lian Chen Wei Ming Wang Nan Chen De Chang Dong Department of Nephrology Ruijin HospitalShanghai Second Medical UniversityShanghai 200025China Animal Laboratory Ruijin HospitalShanghai Second Medical University Department of Gastroenterology Ruijin HospitalShanghai Second Medical University

AIM To evaluale the potential role of p-selectinand anti-p-selectin monoclonal antibody(mAb)in apoptosis during hepatic/renal ischemia-reperfusion injury.METHODS plasma p-selectin level,hepatic/renal p-selectin expression and cell apoptosiswere detected in rat model of hepatic/ renalischemia-reperfusion injury.ELISA,immunohist-ochemistry and TUNEL were used.Someischemia-reperfusion rats were treated with anti-p-selectin mAb.RESULTS Hepatic/renal function insuffic-iency,up-regulated expression of p-selectin inplasma and hepatic/renal tissue,hepatic/renalhistopathological damages and cell apoptosiswere found in rats with hepatic/renal ischemia-reperfusion injury,while these changes becameless conspicuous in animals treated with anti-p-selectin mAb.CONCLUSION p-selectin might mediateneutrophil infiltration and cell apoptosis andcontribute to hepatic/renal ischemia-reperfusioninjury,anti-p-selectin mAb might be an efficientapproach for the prevention and treatment ofhepatic/renal ischemia-reperfusion injury.

关键词： hepatic ischemia reperfusion injury renal ischemia reperfusion injury p-selectin antibody monoclonal apoptosis

Fibrinogen and p-selectin expression in atherosclerosis model of Sprague Dawley rat

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Chinese Medical Journal 2011年第22期124卷 3768-3772页

作者： ZHOU Bi-rong pAN Ying ZHAI Zhi-min Department of Hematology Second Affiliated Hospital of Anhui Medical University Hefei Anhui 230601 China

Background platelet p-selectin plays an important role in inflammation and contributes to thrombosis and hemostasis. Fibrinogen may take part in inflammation, thrombosis, and hemostasis via enhancement of platelet p-selectin expression. This study aimed to discover the correlation between them in atherosclerosis model of Sprague Dawley （SD） rat. Methods Diet-induced atherosclerosis SD rats were adopted as experimental models. The blood from the common abdominal aorta of the rats was obtained to measure the biochemical characteristics and for the check of flow cytometry. Then the aortas were separated carefully, taken out, put into 10% （w/v） neutral formalin for later use. Then fibrinogen and p-selectin expression were detected by flow cytometry and immunohistochemistry. Results SD rats were induced to atherosclerosis model by high fat diet and vitamin D2 injected. It was discovered that the binding of fibrinogen and the expression of p-selectin on the platelet increase in atherosclerosis model （Group H） than in that in the control group （Group Z）, there were closely interrelated. High levels of fibrinogen and p-selectin express on the artery of atherosclerosis rat model. Conclusions Fibrinogen and p-selectin are concerned with atherosclerosis. Fibrinogen can interact with p-selectin in order to contribute to the development of atherosclerosis, high levels of fibrinogen and p-selectin can be regarded as risk factors for markers of atherosclerosis.

关键词： fibrinogen p-selectin atherosclerosis

Relationship between platelet p-selectin and severity of acute coronary syndromes

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Journal of Geriatric Cardiology 2008年第3期5卷 146-149页

作者： Rui Wang~1 Jiyuan Lu~1 Yongping Jia~1 Yuping Gao~1 Chunyu Fan~1 Fang Li~2 1 Department of Cardiology, The First Hospital of Shanxi Medical University. 2 Department of Biochemisry and Molecular Biology, Shanxi Medical University. Taiyuan, Shanxi 030001, China Department of Cardiology First Hospital Shanxi Medical University 85 Jiefang South Rode Taiyuan 030001 Shanxi China Department of Biochemisry and Molecular Biology Shanxi Medical University Taiyuan Shanxi 030001 China

Objective Recent studies reveal important roles ofplatelet p-selectin on progression of atherosclerosis.In the present study, we examine the relation between platelet p-selectin expression and severity of acute coronary syndromes.Methods One hundred and eighty-four consecutive patients with proven or clinically suspected acute coronary syndromes (ACS) were enrolled in the study. Level of p-selectin expression was determined by flow cytometry.platelet p-selectin level was expressed as the percentage of p- selectin positive platelet.Results The level of p-selectin was higher in patients with a single diseased coronary artery or multiple diseased arteries compared to those with normal coronary arteries.p-selectin expression was significantly and positively correlated with angiographic Gensini score (r=0.323,p=0.029).Multiple regression analyses showed that the association of the percentage of p- selectin-positive platelets with ACS was independent of other clinical factors.Conclusions platelet p-selectin is associated with severity of acute coronary syndromes in patients with acute coronary syndromes.

关键词： platelets p-selectin acute coronary syndromes

探讨丹红注射液对早期急性脑梗死血管再通治疗后p-selectin,vWF的影响

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探讨丹红注射液对早期急性脑梗死血管再通治疗后P-selectin,vWF的...

作者：关卓杰湖南中医药大学

学位级别：硕士

目的:探讨丹红注射液对急性脑梗死血管再通治疗后大鼠vWF、p-selectin的影响及安全性,为临床AIS静脉溶栓后的早期应用提供新的思路和科学依据。方法:将60只大鼠随机分为假手术组、梗死组、溶栓组、丹红注射液组,每组再分为1d、3d、7d三... 详细信息

目的:探讨丹红注射液对急性脑梗死血管再通治疗后大鼠vWF、p-selectin的影响及安全性,为临床AIS静脉溶栓后的早期应用提供新的思路和科学依据。方法:将60只大鼠随机分为假手术组、梗死组、溶栓组、丹红注射液组,每组再分为1d、3d、7d三个时间亚组,每个时间亚组5只动物,每组各15只大鼠。假手术组在1h、1.5h及每日后予以等体积0.9%氯化钠溶液静脉注射;梗死组在梗死后1h、1.5h及每日后予以等体积0.9%氯化钠溶液静脉注射;溶栓组在梗死后1h静脉注入阿替普酶（10mg/kg）,1.5h及每日后给药同梗死组;丹红组在梗死后1h同溶栓组,溶栓后30min及每日还序贯静脉注入丹红注射液（5ml/kg/d）。计算各组大鼠死亡率,在相应时间点完善大鼠神经功能缺损评分,然后采血,断头取脑,TTC染色观察梗死面积,HE染色观察皮层神经元形态,ELISA法检测血清中p-selectin、vWF浓度。结果:1.死亡率:与假手术组相比,各组大鼠死亡率明显升高（p（27）0.05）;其余各组之间两两比较,死亡率均无明显差异（p（29）0.05）。2.神经功能缺损评分:与假手术组比较,各组在不同时间点的评分均明显升高（p（27）0.05）;与梗死组比较,溶栓组及丹红组在不同时间点的评分均明显降低（p（27）0.05）;与溶栓组比较,丹红组在7d时的评分明显降低（p（27）0.05）,但在1d、3d时的评分无显著差异（p（29）0.05）。3.TTC染色:与假手术组比较,梗死组、溶栓组、丹红组在各时间点的梗死面积均明显增加（p（27）0.05）;与梗死组比较,溶栓组、丹红组在各时间点的梗死面积均明显减小（p（27）0.05）;与溶栓组比较,丹红组在3d、7d时的梗死面积明显减小（p（27）0.05）,但在1d时的梗死面积无显著差异（p（29）0.05）。4.HE染色:假手术组各时间点大鼠神经元结构清晰,排列整齐,细胞核淡染;梗死组可见神经元萎缩变形,变性坏死,核仁固缩,胞质深染,细胞周围间隙增大,部分细胞水肿;与梗死组相比,溶栓组及丹红组细胞空泡样变,核仁皱缩,变性坏死明显减轻,胶质细胞增生减少,细胞周围间隙增宽减小;丹红组较溶栓组细胞排列更加整齐。5.血清p-selectin浓度:（1）与假手术组比较:梗死组、溶栓组在1d、3d、7d时的p-selectin浓度均明显升高（p（27）0.05）;丹红组在1d时的p-selectin浓度明显升高（p（27）0.05）,在3d、7d时的p-selectin浓度无明显差异（p（29）0.05）。（2）与梗死组比较:丹红组在1d、3d、7d时的p-selectin浓度均明显降低（p（27）0.05）;而溶栓组在1d、3d、7d时的p-selectin浓度均无明显差异（p（29）0.05）。（3）与溶栓组比较:丹红组在1d、3d、7d时的p-selectin浓度均明显降低（p（27）0.05）。血清vWF浓度:（1）与假手术组比较:梗死组、溶栓组在1d、3d、7d时的vWF浓度均明显升高（p（27）0.05）;丹红组在1d时的vWF浓度明显升高（p（27）0.05）,在3d、7d时的vWF浓度无明显差异（p（29）0.05）。（2）与梗死组比较:丹红组在1d、3d、7d时的vWF浓度均明显降低（p（27）0.05）;而溶栓组在1d、3d、7d时的vWF浓度均无明显差异（p（29）0.05）。（3）与溶栓组比较:丹红组在1d、3d、7d时的vWF浓度均明显降低（p（27）0.05）。结论:1.丹红注射液对早期急性脑梗死静脉溶栓模型大鼠的神经功能缺损症状有明显改善,且在早期应用是安全的。2.丹红注射液能够通过降低早期急性脑梗死静脉溶栓模型大鼠血清中的vWF、p-selectin水平,从而抑制血小板的活化,改善神经功能。

关键词：急性脑梗死血管再通丹红注射液 p-selectin vWF

Significance　of　p-selectin　expression　in　human　glomerulonephritis

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Chinese Medical Journal 1997年第7期 16-18页

作者：李晓周同郝翠兰董德长程砜陈诗书

Objective To examine the relationship between p selectin expression and human glomerulonephritis. Methods p selectin expression was investigated in renal biopsies of glomerulonephritis patients (n=133) by immunohistochemical and in situ hybridization analyses. They were divided into three groups based on the degrees of cells proliferation or sclerosis. Group Ⅰ: histological lesions in glomeruli are mild (n=23); Group Ⅱ: glomerular cells proliferation is prominent (n=91); Group Ⅲ: glomerular sclerosis is severe (n=19). The Kruskal Wallis test, Chisquare test and Spearman's correlation were used in our study. Results In normal controls, renal p selectin expression was negative (n=10). In glomerulonephritis, the up regulated p selectin expression on tubular epithelium was significantly higher than that on glomeruli and interstitium (pp selectin was expressed predominantly on platelets in glomeruli, and glomerular p selectin expression was more significantly up regulated in Group Ⅱ than in Group Ⅰ or Group Ⅲ (pppectively). There were strong correlations between the degree of tubulointerstitial lesions and the expression of p selectin on tubular epithelium or within interstitium (rs=0.395 and rs=0.337, pp slectin messenger RNA (mRNA) signals were also detected in the cytoplasm of glomerulus, tubular epithelial cells, intertitium and vascular endothelium. Conclusions p selectin might mediate intraglomerular platelets aggregation, activation and leukocyte accumulation in the early stages of human proliferative glomerulonephritis. The up regulation of p selectin in interstitium was associated with interstitial fibrosis and tubular atrophy and may contribute to the progression of glomerulonephritis.

关键词：德长 expression Significance glomerulonephritis human 周同 p-selectin