Roles Of CD147 And RECK In The Pathogenesis And Therapy Of Systemic Lupus Erythematosus

BackgroundSystemic lupus erythematosus(SLE)is the most representative autoimmune disease in humans.It is characterized by the presence of activated T cells and B cells in conjunction with the development of many different auto antibodies,and chronic inflammation that can affect various parts of the body including skin and kidney,resulting in significant morbility and mortality.Despite a large number of studies,the etiology and pathogenesis of SLE are not completely understood yet.An extremely complicated and multifactorial interaction among various genetic,hormone and environmental factors is probably involved.As a basic pathology change happened in SLE,vasculitis appears in about fifty percents of SLE patients with various clinical manifestations.It could appear at any stage of SLE and has been estimated as the most effective index of disease activity.It is most commonly recognized in the skin but may be responsible for symptoms and findings in a number of organs and its presence is associated with increased mortality.For example, pulmonary hypertension will happen if vasculitis appears in lung;acute pancreatitis or haemorrhage,perforation or necrosis of esophagus,stomach,intestine or colon will happen if vasculitis appears in digestive system;crisis of lupus encephalopathy including anepia,hemiparalysis or major epilepsy caused by cerebral infarction or cerebral haemorrhage will happen ifvasculitis appears in central nervous system.The prognosis of patients with SLE has improved substantially over the past two decades,with a ten-year survival of more than ninety percents in some centers.The prognosis heterogeneity associated with SLE can be attributed to gender,age,genetic clinical factors and treatment.Education,earlier intervention,better disease damage control and improved general management of hypertension,co-morbid infections and other clinical risks have contributed to reduced end-organ damage and less morbidity and mortality.Nonetheless,the serious organ damage caused by vasculitis remains a challenge for physicians.Further study of the pathology and controlling methods of vasculitis in SLE helps to find new strategies to improve long-term prognostic and life quality of the SLE patients.Members of matrix metalloproteinases(MMPs)family can degenerate components of extracellular cell matrix(ECM)and thus play an important role in vasculitis.As a member of matrix metalloproteinases family,matrix metalloprteinases-9(MMP-9)has been associated with SLE for several years since 2002.It cleaves denatured collagens and type IV collagen,the major component of the basement membranes,and thus be involved in many physical and pathological progresses such as embryonic development,repair in trauma,vasculogenesis,inflammation,infiltration and metastasis of tumor and connective tissue diseases.MMP-9 triggers inflammation directly,by tissue destruction,or indirectly, by generation of an inflammatory signal or recruitment of inflammatory cells.Thus, overproduced MMP-9 may induce microvascular damage and leakage of substances that further augment endothelial cell damage and facilitate the movement of inflammatory cells across the basement membrane,ultimately leading to various vasculities of SLE. Inhibition of MMP-9 might be beneficial to preventing organ damage in SLE caused by vasculitis.However,studies on changes of serum concentration of MMP-9 in SLE and the correlation between serum concentration of MMP-9 and disease activity showed different results.Because recta-analysis is an efficient tool to systematically review those published articles in associated articles with different results which could lead to powerful evidence and right direction for further research,we collected the published articles and make meta-analysis of the serum MMP-9 concentration changed in SLE patients and the differences between patietns with different disease activity.In recent years,some MMPs inhibitors such as KR-31813 and BB-1101 were studied in therapy of certain human cancer and autoimmune diseases but no clinical efficacy was demonstrated.The non-specificity of inhibition or serious skeletal muscle damage of articular damage of these inhibitors calls for a new inhibition of MMPs.A new therapy targeted on MMP-9 which can improve long-term prognostic and life quality of patients with SLE is expected. CD147,also known as tumor cell-derived collagenase stimulatory factor(TCSF), extracellular matrix metalloproteinase inducer(EMMPIN),human leukocyte activated antigen M6,and basic immunoglobulin(Basigin),is a member of immunoglobulin superfamily(IgSF).It is capable of inducing the expression and activation of several MMPs of different cells and involves in intercellular action and interaction between cells and matrix thus play an important role in infiltration and metastasis of tumor and development of some autoimmune diseases by an as yet unknown mechanism.CD147 is highly expressed on activated T lymphocytes and was identified as an activation associated antigen on phytohemagglutinine(PHA)activated T cells.Reversion-inducing cysteine-rich protein with Kazal motifs(RECK),a novel MMPs inhibitor,is a membrane-anchored glycoprtein of approximately 110kD that contains multiple epidermal growth factor-like repeats and serine protease inhibitor-like domains. It inhibits MMP-2,MMP-9 and membrane type-1 secretion and activity.As a new MMPs inhibitor,RECK has been shown to be a prognostic marker in several kind of human tumor and plays an important role in angiogenesis.The expression of RECK in inflamed synovial membranes of patients with rheumatoid arthritis was decreased compared to those of patients with osculoarthritis or traumatic arthritis.Downregulation of RECK has been complicated with angiogenesis and infiltration of tumor.Apart of these studies,little was known on what role it plays in SLE.ObjectiveTo evaluate changes of serum concentration of MMP-9 in SLE patients and the correlation between serum concentration of MMP-9 with disease activity;determine the expression of CD147,RECK protein,RECK mRNA,MMP-9 mRNA and MMP-9 secretion of PBMCs in SLE patients;analyze correlation between these factors and clinical characters of SLE patients;study the roles of CD147 and RECK played in SLE and evaluate their value in SLE therapy.Objects and methods1.Objects(1)Reported articles about serum concentrations of MMP-9 and correlation between serum concentration of MMP-9 and disease activity in SLE;(2)Sixty nine SLE patients; (3)Twenty three healthy donors.2.Methods(1)Articles searched out were selected critically according to the criteria of Lichtenstein;(2)Peripheral blood mononuclear cells(PBMCs)were isolated by Ficoll gradient centrifugation;(3)Flow cytometry analysis(FCAS)was used to determine the expression of CD147 on PBMCs;(4)Semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)was used to determine the expression of RECK mRNA and MMP-9 mRNA;(5)Western blotting was used to determine the expression of RECK protein;(6)Enzyme linked immunosorbent assay(ELISA)was used to detect the concentrations of MMP-9 in serum and culture supernatants;(7)Statistical analysis①Meta analysis was used to evaluate the changes of serum concentration of MMP-9 in SLE patients and correlation between serum concentration of MMP-9 with disease activity;②Student's t test was used to analysis the differences between between variables in SLE patients and healthy donors;③Pearson r correlation test was used to evaluate the relationship between variables;④A 2×2 factorial experiment was designed to study the effects of anti-CD147 monoantibody on the expression of RECK,the expression of MMP-9 and secretion of MMP-9;⑤P values of less than 0.05 were considered significant.Results1.Meta analysis(1)There is no difference between serum concentrations of MMP-9 in SLE patients and healthy controls;(2)There is no difference between serum concentrations of MMP-9 in SLE patients with different disease activity. 2.Changes of expression of CD147,RECK and MMP-9,MMP-9 secretion of PBMCs and serum concentration of MMP-9 in patients with SLE and their clinical significance(1)CD147 expression of PBMCs from SLE patients and its relationship with clinical featuresCD147 was highly expressed on PBMCs from SLE patients than on PBMCs from healthy donors.PBMCs from SLE patients with oral ulcers,renal damage,or those with abnormal serum concentration of anticardiolipin antibody were more likely to have higher expression of CD147 than those from SLE patients without such manifestation.A positive correlation between CD147 expression and SLEDAI was observed,but no correlation between CD147 expression and SDI was found.(2)RECK expression of PBMCs from SLE patients and its relationship with clinical featuresPBMCs from SLE patients expressed lower RECK protein compared with those from healthy donors.PBMCs from SLE patients with malar rash,oral ulcers,renal damage or those with abnormal serum concentration of anti-Smith antibody or anticardiolipin antibody were more likely to have much lower expression of RECK protein than those from SLE patients without such manifestation.A negative correlation between RECK protein expression and SDI was observed,but no correlation between RECK expression and SLEDAI was found.PBMCs from SLE patients expressed lower RECK mRNA compared with those from healthy donors.PBMCs from SLE patients with discoid rash were more likely to have much lower expression of RECK mRNA than those from SLE patients without discoid rash.No correlation was found between RECK mRNA and SDI.No correlation was found between RECK mRNA and SLEDAI.(3)MMP-9 mRNA expression of PBMCs from SLE patients and its relationship with clinical featuresPBMCs from SLE patients expressed higher MMP-9 mRNA compared with those from healthy donors.PBMCs from SLE patients with oral ulcer or those with abnormal serum concentration of antinuclear antibodies were more likely to have much higher expression of MMP-9 mRNA than those from SLE patients without such manifestation. MMP-9 mRNA expression was positively correlated with SLEDAI.MMP-9 mRNA expression was positively correlated with SDI,too. (4)MMP-9 secretion of PBMCs from SLE patients and its relationship with clinical featuresPBMCs from SLE patients secreted more MMP-9 compared with those from healthy donors.PBMCs from SLE patients with malar rash,oral ulcer,renal damage or those with abnormal serum concentration of anti-Smith antibody or anticardiolipin antibody were more likely to secreted much more MMP-9 than those from SLE patients without such manifestation.MMP-9 secretion was positively correlated with SLEDAI.MMP-9 secretion was positively correlated with SDI,too.(5)Serum concentration of MMP-9 in SLE patients and its relationship with clinical featuresThere is no difference between the MMP-9 serum concentrations of SLE patients and healthy donors.The concentration of MMP-9 in serum of SLE patients with renal damage was lower than those of SLE patients without renal damage.No correlation between serum concentrations of MMP-9 with SLEDAI was found.No correlation between serum concentrations of MMP-9 with SDI was found.3.Correlation between expression of CD 147,RECK,MMP-9,secretion of MMP-9 and serum concentration of MMP-9 in patiens with SLE(1)Correlation between CD 147 expression and MMP-9 expressionCD147 expression was positively correlated with MMP-9 mRNA expression;(2)Correlation between CD147 expression and MMP-9 secretionCD147 expression was positively correlated with MMP-9 secretion;(3)Correlation between CD147 expression and serum concentration of MMP-9No correlation was found between CD147expression and serum concentation of MMP-9;(4)Correlation between RECK expression and MMP-9 expressionThe expression of RECK protein was negatively correlated with MMP-9 mRNA expression,but no correlation was found between the expression of RECK mRNA and MMP-9 mRNA expression;(5)Correlation between RECK expression and MMP-9 secretionThe expression of RECK protein was negatively correlated with MMP-9 secretion, but no correlation was found between the expression of RECK mRNA and MMP-9 secretion;(6)Correlation between RECK expression and serum concentration of MMP-9 Neither of RECK protein expression nor RECK mRNA expression was correlated with serum concentration of MMP-9;(7)Correlation between CD 147 expression and RECK expressionThe expression of CD147 was negatively correlated with RECK protein expression, but no correlation was found between the expression of CD147 and RECK mRNA;4.Effects of anti-CD147 monoantibody on expression of RECK,MMP-9 and secretion of MMP-9(1)As to PBMCs from SLE patients,anti-CD147 monoantibody could increase RECK protein expression,decrease MMP-9 mRNA expression and decrease MMP-9 secretion;(2)As to PBMCs from healthy donors,anti-CD147 monoantibody had no effect on expression of RECK protein,RECK mRNA,MMP-9 mRNA and secretion of MMP-9;(3)Anti-CD147 monoantibody could decrease the effects of PHA(MMP-9 secretion increased,MMP-9 mRNA expression increased,RECK protein expression decresed)on PBMCs from SLE patients and healthy donors.Conclusion1.The serum concentration of MMP-9 should not be used in SLE diagnosis,neither should it be used in judgement of disease activity;2.Compared with PBMCs from healthy donors,PBMCs from SLE patients expressed higher CD147,lower RECK protein,lower RECK mRNA,higher MMP-9 mRNA and secreted more MMP-9;3.PBMCs from SLE patients with oral ulcers,renal damage or those with abnormal serum concentration of anticardiolipin antibody were more likely to express much higher CD147,lower RECK protein,lower RECK mRNA,lower MMP-9 mRNA and secret more MMP-9 than those from SLE patients without such manifestation.Physicians should pay more attention to those patients because serious organ damage caused by vasculitis are more likely to appear;4.CD147 expression on PBMCs from SLE patients was positively correlated with disease activity and could be used to assess disease status and estimate the therapeutic effects;5.RECK protein expression on PBMCs from SLE patients was negatively correlated with damage accrual and might be regarded as an indicator of the disease course allowing clinicians to anticipate the intermediate and long-term outcomes of the disease; 6.MMP-9 mRNA expression was positively correlated with disease activity and damage accrual of SLE patients and could be used to assess disease status and anticipate the prognostics of SLE patients;7.MMP-9 secretion ability of PBMCs in SLE patients was positively correlated with disease activity and could be used to assess disease status and estimate the therapeutic effects;8.There is no difference between serum concentrations of MMP-9 in SLE patients and healthy donors;serum concentration was correlated with neither disease activity nor damage accrual;9.Over expression of CD147 inhibited the expression of RECK protein and this inhibition increased the secretion of MMP-9 which plays important role in SLE pathology;10.The decreased expression of RECK was correlated with overproduction of MMP-9.Vascular damage could be prevented by increasing the expression of RECK protein.This will be benefit for development of new therapy.11.Anti-CD147 monoantibody could inhibit MMP-9 secretion of PBMCs in SLE patients.This effect might be correlated with upregulation of RECK protein.Anti-CD147 monoantibody might be prospective in SLE treatment.