RESUMEN
Objective To explore the mechanism of chitosan oligosaccharides(COS)in reducing atherosclerotic plaque formation from the perspective of protein glycosylation modification.Methods Totally 40 ApoE-/-mice were randomly divided into control group and COS group.The control group was given a high-fat diet for 12 weeks,and COS group was given a high-fat diet plus COS(gavage per day,500 mg/kg)for 12 weeks.Serum lipid detection,HE staining and Oil red O staining were used to detect plaque formation.Lectin chip,liquid chromatography tan-dem-mass spectrometry and ELISA were used to detect potential changes of glycoprotein in serum.Cholesterol ester outflow and free cholesterol ester determination experiment were used to evaluate the effect of changes in scavenger receptor class B type Ⅰ(SRBI)protein glycosylation modification site on cholesterol effluence in macrophages.Results COS significantly reduced the level of TC and LDL-C(P＜0.05)in mice,but had no effect on the level of TG,HDL-C,ApoA1 and ApoB100.The intima thickness and plaque size of the aorta were significantly thinner and smaller(P＜0.05)in the COS group compared with the control group.The molecular weight of lens culinaris ag-glutinin(LCA)binding protein with the most obvious change is 80-90 ku,and SRBI was one of them.COS promo-ted the cholesterol outflow and inhibited the accumulation of free cholesterol ester in RAW264.7 cell(P＜0.05).Knockdown or glycosylation site mutation with SRBI inhibited cholesterol outflow caused by COS,and increased the accumulation of intracellular free cholesterol(P＜0.05).Conclusions COS promotes lipid efflux by increasing SRBI glycosylation and expression,thereby alleviating atherosclerotic plaque formation.
RESUMEN
BACKGROUND:Compound Shengmai Chenggu capsule has good therapeutic effects on early steroid-induced osteonecrosis of the femoral head,but the exact mechanism of treatment is not fully understood. OBJECTIVE:To observe the effect of compound Shengmai Chenggu capsule on fucosyltransferase 8,osteogenic gene and Wnt/β-catenin in bone tissue of rats with steroid-induced osteonecrosis of the femoral head. METHODS:Sixty Sprague-Dawley rats were randomized into blank group,model group,low-,middle-,and high-dose drug groups(n=12 per group).In the latter four groups,animal models of steroid-induced osteonecrosis of the femoral head were established by subcutaneous injection of imiquimod(once every 2 weeks,2 times in total)and gluteal muscle injection of methylprednisolone(once a week,4 times in total).The low-,middle-and high-dose drug groups were given 1.89,3.78 and 7.56 g/kg per day compound Shengmai Chenggu capsule solution by gavage respectively on the second day after the last modeling.The same amount of saline was given by gavage to the model group.Administration lasted 8 weeks.After the administration,micro-CT scan,histological staining,compression test,RT-qPCR and western blot were performed on the femoral head. RESULTS AND CONCLUSION:Micro-CT scan results showed that compared with the blank group,trabecular volume fraction,trabecular number and trabecular thickness were significantly decreased(P<0.05),while trabecular separation was increased in the model group(P<0.05).Compared with the model group,the compound Shengmai Chenggu capsule could increase trabecular volume fraction,trabecular number and trabecular thickness(P<0.05),and decrease trabecular separation(P<0.05)in a dose-dependent manner.Hematoxylin-eosin staining results showed that compared with the model group,the rate of empty bone lacunae was reduced in a dose-dependent group in the low-,middle-,and high-dose compound Shengmai Chenggu capsule groups(P<0.05).Immunohistochemical staining results showed that compared with the blank group,the protein expression of fucosyltransferase 8,Runx2 and bone morphogenetic protein 2 was reduced in the model group(P<0.05);compared with the model group,the compound Shengmai Chenggu capsule increased the protein expression of fucosyltransferase 8,Runx2 and bone morphogenetic protein 2 in a dose-dependent manner(P<0.05).Results from the compression test showed that there was a dose-dependent increase in the maximum load and elastic modulus of the femoral head in the low-,middle-,and high-dose compound Shengmai Chenggu capsule groups compared with the model group(P<0.05).RT-qPCR and western blot results showed that the mRNA and protein expressions of fucosyltransferase 8,Runx2,alkaline phosphatase,osteocalcin,osteoblast-specific transcription factor and bone morphogenetic protein 2 were decreased in the model group compared with the blank group(P<0.05);compared with the model group,there was a dose-dependent increase in the mRNA and protein expressions of the above indicators in the low-,middle-,and high-dose compound Shengmai Chenggu capsule groups compared with the model group(P<0.05).Compared with the blank group,the mRNA and protein expression of Wnt2,low-density lipoprotein receptor-related protein 5 and β-catenin were decreased(P<0.05)and the mRNA and protein expressions of glycogen synthase kinase 3β were increased(P<0.05)in the model group;compared with the model group,there was a dose-dependent increase in the mRNA and protein expressions of Wnt2,low-density lipoprotein receptor-related protein 5 and β-catenin(P<0.05)but a dose-dependent decrease in the mRNA and protein expressions of lycogen synthase kinase 3β(P<0.05)in the low-,middle-,and high-dose compound Shengmai Chenggu capsule groups.To conclude,the mechanism by which the compound Shengmai Chenggu capsule treats steroid-induced osteonecrosis of the femoral head may activate the Wnt/β-catenin signaling pathway through the up-regulation of fucosyltransferase 8,thereby promoting bone formation.
RESUMEN
Objective To establish a platform for enrichment and purification of core fucose glycosylation and analyse its immu‐nity properties .Methods Serum immunoglobulin G(IgG) was seperated and purified by using protein G cross‐linked sepharose ,and core fucose glycosylation components of IgG were enriched and purified by using lens culinaris agglutinin (LCA)affinity chromatog‐raphy ,meanwhile ,the isoforms of LCA‐IgG were determined by using Western blotting .Results The purification rate of serum IgG was 62% and yield rate of LCA‐IgG was 4 .59% .Serum IgG1 ,IgG2 and IgG3 were identified except IgG4 in patients with schizo‐phrenia ,while serum IgG1 ,IgG2 ,IgG3 and IgG4 were all identified in healthy individuals .Conclusion A method for enrichment and purification of core fucose glycosylation in patients with schizophrenia have been successfully established and distribution character‐istics of the LCA‐IgG isoforms have been determined ,which could provide a new insight into immune pathomechanism of schizo‐phrenia .
RESUMEN
Objective To investigate the correlation between the levels of serum immunoglobulin core fucosylation and humoral immune parameters in patients with schizophrenia and its immunological significance .Methods The levels of serum lens culinaris agglutinin(LCA)-IgM ,LCA-IgG ,LCA-IgA ,IgM ,IgG ,IgA ,circular immune complex(CIC) in patients with schizophrenia(n= 46) and health people(n=47) were determined .Their correlations were investigated by linear regression analysis .Results There was a positive correlation between the LCA-IgA and IgG in healthy people(r=0 .311 7 ,P<0 .05) .However ,there was a negative correla-tion between LCA-IgG and IgA in patients with schizophrenia(r=0 .323 6 ,P<0 .05) .In addition ,there were positive correlations between LCA-IgM ,LCA-IgG ,LCA-IgA and CIC(r=0 .354 8 ,r=0 .189 8 ,r=0 .479 7 ,P<0 .05) .Conclusion Increased fucosyla-tion levels of immunoglobulin were detected in patients with schizophrenia ,which may play important roles in immunological inju-ries of central nervous system in schizophrenia .
RESUMEN
A glycomic method was used to screen the aberrantly ?1-6 fucosylated glycoproteins related to HCC metastasis and analyze the alteration of CK8 both in its expression level and its glycan parts associated with metastatic ability. Based on the approach, 2-DE coupled with lectin affinity blot, lectin affinity precipitation followed by MALDI-TOF-MS/MS, the lens culinaris agglutinin (LCA) affinity glycoprotein profiles from MHCC97-L and MHCC97-H cells, two higher metastatic HCC cell lines, were obtained, in which a differentially displayed protein spot was indicated when compared with Hep3B, in the region within 55～60 ku in molecular mass and 4～6 in isoelectric point. The identification result was CK8 by MALDI-TOF-MS/MS. To confirm the relation between increased core-fucosylation of CK8 and HCC metastasis, LCA affinity precipitation was used to extract the ?1-6 fucosylated glycoproteins, followed by Western blot. And it was found that CK8 was highly fucosylated in both MHCC97-L and MHCC97-H cells compared to Hep3B. Immunofluorescence analysis and Western blot were used to detect its intracellular localization and its protein expression levels, indicating that CK8 distributed in cytoplasm and increased protein expressions in MHCC97-L and MHCC97-H cell lines. And further lectin binding studies found that CK8 has a high affinity for Con A in both MHCC97-H and Hep3B cells, indicating that CK8 was a glycoprotein with high-mannose type N-glycans. But the amount of the lectin RCA-1 binding to CK8 was greater in MHCC97-H than Hep3B, suggesting that CK8 contained the increased terminal galactose residues ?-1, 4-linked to GlcNAc in MHCC97-H. All the results suggested that the increase of CK8 in its protein expression level, core-fucosylation and terminal gal ?1,4 GlcNAc disaccharides might be related to HCC metastatic ability.