1.Effects of combination of dihydroartemisinin and Huobahua on expression of p-p38 MAPK and ICAM-1 in mice with delayed-type hypersensitivity.
Min-Xia ZHAN ; Du WEI ; Li HENG-HUA ; Jin-Ping LUO ; Xiao-Li MEI ; Tu RU-XIA ; Li ZHANG
China Journal of Chinese Materia Medica 2020;45(22):5561-5566
The aim of this paper was to investigate the immunosuppressive effects of dihydroartemisinin and Huobahua compatibility in mice with delayed hypersensitivity and explore its possible mechanism. The delayed-type hypersensitivity(DTH) model in mice was established to observe the immunosuppressive effects of dihydroartemisinin and Huobahua compatibility in DTH mice. ELISA assay was used to detect the contents of interferon(IFN-γ); histopathological changes and degree of mononuclear infiltration of right ear tissues were examined by HE staining; the expression level of intercellular cell adhesion molecule-1(ICAM-1) in the right ear of mice was detected by immunohistochemistry; the protein expression levels of p38 phospho mitogen activated protein kinase(p-p38 MAPK) was detected by Western blot analysis. As compared with the control group, the degree of ear swelling, thymus/spleen index, serum IFN-γ as well as the number and degree of infiltration of monocytes were significantly increased in the model group. As compared with the model group, the degree of ear swelling and thymus/spleen index of the mice in the combination group were significantly reduced; the number and degree of infiltration of monocytes were significantly relieved; the serum levels of IFN-γ and the expression levels of p-p38 MAPK and ICAM-1 proteins in the right ear were also significantly reduced. The combination of dihydroartemisinin and Huobahua can significantly inhibit the DTH response, and it may regulate the production and secretion of related inflammatory factor IFN-γ by inhibiting the phosphorylation activity of p38 MAPK, thereby further reducing the expression of ICAM-1 and thus exerting the immunosuppressive effect.
Animals
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Artemisinins
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Intercellular Adhesion Molecule-1/genetics*
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Mice
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Monocytes
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p38 Mitogen-Activated Protein Kinases/genetics*
2.A study on K469E polymorphism of ICAM1 gene and ICAM1 plasma level in patients with coronary heart disease.
Fang-hong LU ; Qing SHANG ; Pei-e WEN ; Guo-hai SU ; Jian-mei WU ; Qi TIAN ; Ying-xin ZHAO
Chinese Journal of Medical Genetics 2006;23(2):195-197
OBJECTIVETo study the linkage between K469E polymorphism of intercellular adhesion molecule 1(ICAM1) gene with ICAM1 plasma level and coronary heart disease (CHD) in Han population of China.
METHODSOne hundred and sixty-four controls without CHD and 160 patients with CHD were enrolled in our study. By nested PCR with allele-specific oligonucleotide primers, all patients and controls were genotyped for the ICAM1 polymorphism. And the ICAM1 plasma level was measured by ELISA.
RESULTSIn the patients with CHD, both K allele frequency and the plasma level of ICAM1 were higher than those in control (P<0.05). The individual with K allele had higher plasma level of ICAM1 than that without K allele (344.34+/-128.59 microg/L vs 303.54+/-108.74 microg/L, P=0.008). K allele enhanced the risk of CHD (P<0.01, OR=2.158, 95%CI: 1.250-3.727). There was the K allele cooperation with smoking in influencing the risk of CHD.
CONCLUSIONThere is the polymorphism of ICAM1 K469E gene in Han population of China, and the K allele may be a genetic factor influencing the risk of CHD.
China ; ethnology ; Coronary Disease ; blood ; genetics ; Gene Frequency ; Humans ; Intercellular Adhesion Molecule-1 ; blood ; genetics ; Polymorphism, Genetic ; Potassium ; antagonists & inhibitors
3.Hemangiopoiet in modulates adhesive properties of endothelial cells.
Xiao-Jing WANG ; Yong-Jun LIU ; Shi-Hong LU ; Lin-Hui LIANG ; Han WANG ; Zhong-Chao HAN
Acta Academiae Medicinae Sinicae 2006;28(3):364-367
OBJECTIVETo explore the effect of hemangiopoietin (HAPO) on the adhesive properties of human umbilical vein endothelial cells (HUVEC).
METHODSThe adhesion of HUVEC and the expressions of CD54, CD102, CD106, CD31, CD62E, and CD62P were measured by adhesion assay, flow cytometry, and semi-quantitative RT-PCR.
RESULTSHAPO enhanced the total adherence of HUVEC in a concentration-dependent manner. Flow cytometry analysis revealed that the treatment of HAPO resulted in a significantly increased expression of CD106 and CD62E on HUVEC in a time-dependent manner. When HUVEC were incubated with HAPO for 6 h, the percentage of CD106 + HUVEC and CD62E HUVEC increased about 2.10 folds and 5.84 folds, respectively, compared with control. The time-course of adhesive molecules mRNA expression indicated that the expression of CD106 and CD62E reached at the maximum 1.86 folds and 6.16 folds, respectively, compared with control.
CONCLUSIONHAPO may facilitate the homing of hematopoietic stem/progenitor cells.
Cell Adhesion Molecules ; metabolism ; Cells, Cultured ; E-Selectin ; biosynthesis ; genetics ; Endothelial Cells ; cytology ; Humans ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Proteoglycans ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Umbilical Veins ; cytology ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics
4.Effects of ICAM-1 gene transfection on the adipogenic differentiation of MSCs.
Fen-fen XU ; Heng ZHU ; Ji-de CHEN ; Yuan-lin LIU ; Yu-xiao LIU ; Rong-xiu ZHENG ; Yi ZHANG
Chinese Journal of Hematology 2013;34(5):435-439
OBJECTIVETo explore the effects of ICAM-1 gene transfection on the differentiation of MSCs to adipocytes.
METHODSThe recombinant retroviral expression plasmid MIGR1-ICAM-1 containing full length of mouse ICAM-1 gene was constructed. The constructed plasmid MIGR1-ICAM-1, empty plasmid MIGR1 and packaging plasmid ECOS were transfected into T293 cell lines and then the supernatant generated from T293 cells were used to infect mouse MSCs cell line C3H10T 1/2. The transfective efficiency was determined by inverted fluorescence microscope, real-time PCR and flow cytometry. Furthermore, ICAM-1 overexpressing MSCs (C3H10T 1/2-ICAM-1) and empty vector transfection MSCs (C3H10T 1/2-MIGR1) were cultured in medium with or without induction reagents, Oil-red-O staining was used to detect the lipid accumulation, and the expression of transcriptional factors C/EBPα and PPARγ, which were key factors in the differentiation of MSCs to adipocytes, were tested by real-time-PCR.
RESULTSThe recombinant retrovirus vector containing mouse ICAM-1 gene was successful constructed. After transfection into MSCs cell line C3H10T 1/2, the overexpression ICAM-1 MSCs cell line (C3H10T 1/2-ICAM-1) and control cell line (C3H10T 1/2-MIGR1) were obtained. Furthermore, these two cell lines were treated without or with adipocytic induction reagents, C3H10T 1/2-ICAM-1 showed significantly lower mRNA expression level for C/EBPα [(1.2 ± 0.7), (2.9 ± 0.9)] and PPARγ [(1557.6 ± 70.2), (7547.0 ± 442.2)] when compared with C3H10T 1/2-MIGR1 [(5.8 ± 0.5), (23.0 ± 2.3) and (2453.0 ± 215.6), (9856.3 ± 542.2)](P < 0.05). Moreover, little lipid droplet and decreased quantity of adipocytes were detected in C3H10T 1/2-ICAM-1 [(3.2 ± 0.5)/well, (12.2 ± 3.8)/well] than that in C3H10T 1/2-MIGR1 [(11.2 ± 0.4)/well, (51.3 ± 2.8)/well] (P < 0.05).
CONCLUSIONOverexpression of ICAM-1 in MSCs can inhibit its adipocytic differentiation.
Adipocytes ; cytology ; Animals ; Cell Differentiation ; Cell Line ; Intercellular Adhesion Molecule-1 ; genetics ; Mesenchymal Stromal Cells ; cytology ; Mice ; Transfection
5.The role of adhesion molecules in rat liver allograft immune response.
Feng LIU ; Yu-lan LIU ; Jun-hong SUN
Chinese Journal of Hepatology 2003;11(8):460-463
Animals
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E-Selectin
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biosynthesis
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genetics
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Graft Rejection
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immunology
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pathology
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Intercellular Adhesion Molecule-1
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biosynthesis
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genetics
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Liver Transplantation
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immunology
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RNA, Messenger
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biosynthesis
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genetics
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Rats
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Rats, Inbred Lew
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Vascular Cell Adhesion Molecule-1
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biosynthesis
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genetics
6.Polymorphisms of intercellular adhesion molecule-1 gene K469E and platelet endothelial cell adhesion molecule-1 gene C373G in patients with preeclampsia.
Yanhong HE ; Zhijian WANG ; Jin JIN ; Xiaowen ZHU ; Yuying CHEN ; Shuying CHEN ; Yanwen CAO
Journal of Southern Medical University 2014;34(4):552-559
OBJECTIVETo investigate the distributions of genotypic and allelic frequencies of intercellular adhesion molecule-1 (ICAM-1) gene K469E and platelet endothelial cell adhesion molecule-1 (PECAM-1) gene C373G in patients with preeclampsia.
METHODSPolymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) and DNA sequencing were used for detecting ICAM-1 gene K469E and PECAM-1 gene C373G genotypes in 110 women with preeclampsia and 110 normotensive pregnant women in comparison with their clinical characteristics.
ESULTSThe distributions of observed and expected genotype frequencies were consistent with Hardy-Weinberg equilibrium. No significant differences were found in the genotype and allele frequencies of ICAM-1 gene K469E between the two groups (P>0.05), but the CC and the CG genotype frequencies of PECAM-1 gene C373G were significantly different between them (P<0.05). The relative risk for preeclampsia of CG genotype was 1.959 folds of that in CC genotype carriers (OR=1.959, 95%CI: 1.090-3.520, P=0.024), and this association still existed after adjustment for age, gravidity, parity and BMI in logistic regression models. The C373G allele frequencies showed no significant difference between the two groups (P>0.05).
CONCLUSIONSThe CG genotype of PECAM-1 gene C373G genetically predispose the carriers to preeclampsia, while ICAM-1gene K469E polymorphisms is not associated with preeclampsia.
Adult ; Case-Control Studies ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Platelet Endothelial Cell Adhesion Molecule-1 ; genetics ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Pre-Eclampsia ; genetics ; Pregnancy ; Sequence Analysis, DNA
7.Study on anti-angiogenesis effect of three curcumin pigments and expression of their relevant factors.
Yan-fen HUANG ; Xue-xin ZHU ; Zhi-shan DING ; Gui-yuan LV
China Journal of Chinese Materia Medica 2015;40(2):324-329
To study the in vitro anti-angiogenesis effect of three curcumin pigments (curcumin, demethoxycurcumin, bisdemethoxycurcumin). In the study, the inhibitory effect of the three curcumin pigments on proliferation of HUVEC cells induced by OX-LDL and the effect on migration of HUVEC cells were detected. The effect on neovascularization was observed by chorioallantoic membrane (CAM) test. The effect on cell adhesion factors ICAM-1 and VCAM-1 of HUVECs were tested by Real-time RT-PCR. It was found that the three curcumins could inhibit the proliferation of HUVEC cells induced by OX-LDL within the dosage range 4, 8, 16 mg x L(-1), with a dose-dependence. The proliferative effect of curcumins on HUVECs was greater than the other two derivatives (P < 0.01). All of the three curcumin pigments inhibited the migration of HUVEC cells and the angiogenesis of chick chorioallantoic membrane (CAM). The migration inhibition rate of curcumins at middle and high concentrations was greater than the other two (P < 0.01). All of the three curcumin could down-regulate the expression of VEGF and ICAM-1, and curcumins showed more obvious effect in down-regulating VEGF than demethoxycurcumin and bisdemethoxycurcumin(P < 0.01); Bisdemethoxycurcumin showed the most significant effect in down-regulating ICAM-1 (P < 0.01). All of the three showed no remarkable effect on expression of VCAM-1, and only bisdemethoxycurcumin showed the down-regulating effect (P < 0.05). According to the findings, all of the three curcumin pigments could resist angiogenesis by inhibiting proliferation and migration of endothelial cells and down-regulating the expression of VEGF and adhesion molecules ICAM-1.
Angiogenesis Inhibitors
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pharmacology
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Animals
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Cell Movement
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drug effects
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Cells, Cultured
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Chorioallantoic Membrane
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drug effects
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Curcumin
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analogs & derivatives
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pharmacology
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Humans
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Intercellular Adhesion Molecule-1
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genetics
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RNA, Messenger
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analysis
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Vascular Cell Adhesion Molecule-1
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genetics
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Vascular Endothelial Growth Factor A
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genetics
8.Gene expression of adhesion molecules in pulmonary and hepatic microvascular endothelial cells during sepsis.
Rongqian WU ; Yingxin XU ; Xuhua SONG ; Xianjun MENG
Chinese Journal of Traumatology 2002;5(3):146-150
OBJECTIVETo study the gene expression of adhesion molecules in pulmonary and hepatic microvascular endothelial cells during sepsis in mice.
METHODSMale mice were subjected to cecal ligation and puncture (CLP) and microvascular endothelial cells in pulmonary and hepatic tissues were harvested at 3 hours (early sepsis) and 12 hours (late sepsis) after CLP, respectively. Gene expression of the adhesion molecules was assessed by reverse transcription polymerase chain reaction (RT-PCR). Simultaneously, the alterations of myeloperoxidase (MPO) activity in pulmonary and hepatic tissues were also examined.
RESULTSE-selectin mRNA levels markedly increased at 3 hours after CLP in both pulmonary and hepatic microvascular endothelial cells, then they returned to the normal level at 12 hours after CLP. Increases in intercellular adhesion molecule-1 (ICAM-1) mRNA levels were found at 3 hours after CLP in both pulmonary and hepatic microvascular endothelial cells, and these levels became higher at 12 hours after CLP. Adhesion molecule-1 (VCAM-1) mRNA expression of vascular cells also increased significantly at 3 hours and 12 hours after CLP in both pulmonary and hepatic microvascular endothelial cells. The level of VCAM-1 mRNA in hepatic microvascular endothelial cells was higher at 3 hours than that at 12 hours after CLP, while the level of VCAM-1 mRNA in pulmonary microvascular endothelial cells was higher at 12 hours than that at 3 hours after CLP. The MPO activity in pulmonary and hepatic tissues increased at 3 hours after CLP, compared with that of the sham group. They both declined significantly at 12 hours after CLP, but they were still higher than that of the sham group.
CONCLUSIONSThe up-regulation of the gene expression of adhesion molecules in pulmonary and hepatic microvascular endothelial cells is an important step for the migration and accumulation of leukocytes at the site of inflammation, which plays a critical role in organ damage during sepsis. And the contribution of the heterogeneity of endothelial cells in organs' vulnerability during sepsis is worth a further investigation.
Animals ; Endothelium, Vascular ; cytology ; Gene Expression ; Intercellular Adhesion Molecule-1 ; genetics ; Liver ; cytology ; Lung ; cytology ; Male ; Mice ; Peroxidase ; metabolism ; Sepsis ; metabolism ; Up-Regulation ; Vascular Cell Adhesion Molecule-1 ; genetics
9.Construction of ICAM-1-GFP and its binding with Molt-4 cells.
Wei-Hua CHEN ; Wan-Ming DA ; Chun-Ji GAO
Journal of Experimental Hematology 2009;17(3):650-655
This study was aimed to clone human intercellular adhesion molecule-1 (ICAM-1) gene, to transfect the constructed eukaryotic expression vector ICAM-1-GFP into CHO cells, as well as to detect ICAM-1-GFP expression in CHO cells binding with Molt-4 cells. ICAM-1 cDNA gene was amplified by RT-PCR and inserted in PMD(18)-T vector. Then ICAM-1 cDNA from pMD18-ICAM-1 vector was subcloned into eukaryotic expression vector pEGFP-C1 to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing were used to confirm the recombinant vector. After stable transfection of CHO-K1 cells with the recombinant vector, the expression and subcellular localization of ICAM-1-GFP were detected by RT-PCR, flow cytometry and fluorescence microscopy. The function of ICAM-1-GFP fusion protein was assessed by the binding of ICAM-1-GFP/CHO cells to Molt-4 cells. The results showed that 1622 bp full-length ICAM-1 cDNA obtained and was successfully ligated with pMD(18)-T-vector, subcloned to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing indicated that recombinant ICAM-1-GFP was successfully constructed and ICAM-1-GFP was expressed stably in CHO cells. ICAM-1-GFP expression was only observed in the cytoplasm of ICAM-1-GFP/CHO cells by fluorescence microscopy. The ICAM-1-GFP/CHO cells were bound to PMA-treated Molt-4 cells. The expression of MEM-148 was very weak in PMA-treated Molt-4 cells. It is concluded that the ICAM-1-GFP eukaryotic expression vector has been constructed successfully and expresses stably in CHO cells. PMA can increase the binding of Molt-4 cells to ICAM-1-GFP/CHO cells by inducing specialized form of ICAM-1 clustering.
Animals
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CHO Cells
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Cricetinae
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Cricetulus
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DNA, Complementary
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genetics
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Genetic Vectors
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Green Fluorescent Proteins
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genetics
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Humans
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Intercellular Adhesion Molecule-1
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genetics
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Recombinant Proteins
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genetics
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Transfection
10.Hepatocellular glycogen alleviates hepatic ischemia reperfusion injury and its relationship to ICAM-1 gene expression.
Li-jun TANG ; Fu-zhou TIAN ; Tao WANG ; Jian-feng CUI ; Hao LUO ; Dong-xuan LI ; Li SHI ; Tao CHEN ; Shu ZOU
Chinese Journal of Hepatology 2008;16(11):858-860
OBJECTIVETo investigate if higher hepatocellular glycogen contents can alleviate hepatic ischemia reperfusion injury and its relationship to ICAM-1 gene expression in hepatic sinusoidal cells (HSCs).
METHODSTwenty-one rabbits fed with a standard diet were randomly divided into three groups (n=7 in each). All the animals were subjected to hepatic ischemia reperfusion injury then sacrificed. Before the injury, group A rabbits fasted for 24 hours; group C rabbits had 6 intravenous glucose solution (25%, 20 ml) injections, 4 hours between two injections. Hepatic enzymological changes, hepatic ICAM-1 mRNA expressions and leukocytic counts in the sinusoids were observed.
RESULTSThe liver glycogen contents of the three groups were significantly different. Livers of group A had higher contents of glycogen (9.85+/-0.91 mg/g. wet tissue); in group B they were 38.93+/-5.72; and in group C they were 48.31+/-6.58. Group C animals had the slightest liver function damage. There were no differences in the pre- and post-ischemic ICAM-1 mRNA contents in the three groups. However, livers with a higher content of glycogen showed less expression of ICAM-1 mRNA (group A: 1.398+/-0.365 ng/mg wet tissue; group B: 0.852+/-0.297; group C: 0.366+/-0.183) and lower leukocytic counts. The relationship analysis showed a negative relationship between hepatocellular glycogen and hepatic ICAM-1 mRNA contents (r= -0.965, P less than 0.01).
CONCLUSIONSHepatocellular glycogen is important in protecting liver ischemic reperfusion injury. Also hepatocellular glycogen decreases the expression of ICAM-1 mRNA of HSCs.
Animals ; Female ; Glycogen ; pharmacology ; Hepatocytes ; chemistry ; metabolism ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Liver ; chemistry ; metabolism ; pathology ; Male ; RNA, Messenger ; genetics ; Rabbits ; Reperfusion Injury ; genetics ; metabolism ; pathology