OBJECTIVESTo observe the expression of macrophage migration inhibition factor (MIF) and p27 in hepatocellular carcinoma tissue, and to investigate the effect of MIF on the expression of p27 in hepatocellular carcinoma (HCC) cells.

METHODSImmunohistochemistry and quantitative RT-PCR were performed to detect the expression of MIF and p27 in HCC tissues and peri-tumor tissues. Specific small interfering RNA (siRNA) targeting MIF gene was chemically synthesized and then transfected at the concentration of 50 nmol/L and 100 nmol/L into PLC cells and Hep3B cells. The mRNA levels of MIF and p27 after MIF siRNA treatment were quantified by real-time RT-PCR.

RESULTSMIF protein and mRNA were over-expressed in the HCC tumor tissues compared to these in the peri-tumor tissues (P less than 0.01). The expression of p27 protein and mRNA was significantly lower in the HCC tumor tissues compared to these in the peri-tumor tissues (P less than 0.01). Compared to normal liver cell line L-02, HCC cell lines expressed higher level of MIF (F=61.036, P less than 0.01) and lower level of p27 (F=529.853, P less than 0.01). In MIF siRNA treated PLC and Hep3B cells, the MIF mRNA was decreased in a dose-dependent manner (F=320.1, P less than 0.01; F=201.2, P less than 0.01). The p27 mRNA was significantly up-regulated in MIF siRNA treated PLC and Hep3B cells compared to control siRNA transfected cells (F=419.4, P less than 0.01; F=459.9, P less than 0.01).

CONCLUSIONSMIF is over-expressed in HCC tumor tissues, and the expression of p27 is repressed by MIF.

Carcinoma, Hepatocellular ; Cell Line, Tumor ; Humans ; Immunohistochemistry ; Liver Neoplasms ; Macrophage Migration-Inhibitory Factors ; RNA, Messenger ; genetics

OBJECTIVETo investigate the relationship between the plasma macrophage migration inhibitory factor (MIF), activator protein-1 (AP-1) and MMP-9 concentrations and the severity of coronary artery lesions in coronary heart disease (CHD) patients.

METHODSPatients were divided into normal controls (n = 35), stable angina pectoris (SAP, n = 32) and acute coronary syndrome (ACS, n = 75) according to the coronary angiography (CAG), clinical and laboratory examinations. The CAG severity and extent of coronary lesions were analyzed by means of Gensini coronary score system. Enzyme linked immunosorent assay was used to measure the plasma MIF, AP-1 and MMP-9 concentrations.

RESULTSPlasma MIF, AP-1 and MMP-9 concentrations were significant increased in CHD patients [MIF: (14.97 +/- 2.11) microg/L, AP-1: 1.43 +/- 0.33, MMP-9: (1.48 +/- 0.14) microg/L] compared to those in control group [MIF: (9.07 +/- 1.28) microg/L, AP-1: 0.71 +/- 0.13, MMP-9: (1.01 +/- 0.07) microg/L, all P < 0.05]. The MIF, AP-1 and MMP-9 concentrations in ACS group [MIF: (16.66 +/- 2.56) microg/L, AP-1: 1.56 +/- 0.22, MMP-9: (1.58 +/- 0.14) microg/L] were also significant higher than those in SAP group [MIF: (11.01 +/- 2.12) microg/L, AP-1: 1.04 +/- 0.25, MMP-9: (1.25 +/- 0.07) microg/L, all P < 0.05] and there was significant positive correlation between MIF, AP-1 and MMP-9 concentrations and the Gensini score of coronary artery lesions (all P < 0.05). AP-1 was positively correlated with MMP-9 in CHD patients (P < 0.05).

CONCLUSIONSPlasma MIF, AP-1 and MMP-9 concentrations were positively correlated to the severity of coronary lesions in CHD patients. Higher MIF, AP-1 and MMP-9 concentrations in ACS patients than in SAP patients might suggest higher plaque instability in ACS patients.

Coronary Angiography ; Coronary Artery Disease ; blood ; Humans ; Macrophage Migration-Inhibitory Factors ; Plaque, Atherosclerotic

Macrophage migration inhibitory factor (MIF) is originally identified in the culture medium of activated T lymphocytes as a soluble factor that inhibits the random migration of macrophages. MIF is now recognized as a multipotent cytokine involved in the regulation of immune and inf lammatory responses. In rheumatoid arthritis (RA), MIF promotes inf lammatory responses by inducing proinflammatory cytokines and tissue-degrading molecules, promoting the proliferation and survival of synovial fibroblasts, stimulating neutrophil chemotaxis, and regulating angiogenesis and osteoclast differentiation. Expression of MIF in synovial tissue and synovial fluid levels of MIF are elevated in RA patients. Specifically, MIF levels correlate with RA disease activity and high levels are associated with bone erosion. In animal models of RA, the genetic and therapeutic inhibition of MIF has been shown to control inflammation and bone destruction. Based on the role of MIF in RA pathogenesis, small molecular inhibitors targeting it or its receptor pathways could provide a new therapeutic option for RA patients.
Arthritis, Rheumatoid* ; Chemotaxis ; Cytokines ; Fibroblasts ; Humans ; Inflammation ; Macrophage Migration-Inhibitory Factors ; Macrophages* ; Models, Animal ; Neutrophils ; Osteoclasts ; Synovial Fluid ; T-Lymphocytes

In recent years, there has emerged academic tendency towards the neurogenic mechanism of ulcerative colitis (UC). As one of the supports to the hypothesis of UC being a neurogenic inflammation, we have built a colitis model by intrathecal (ith) injection of a haptten 2,4-dinitrochlorobenzene (DNCB) to DNCB-sensitized rats. In order to explore further the neuroimmunal mechanism of this colitis model, we here focused on a pro-inflammatory cytokine, migration inhibitory factor (MIF), to observe its expression in rat colon nervous tissue and spinal cord in the colitis induced by ith injection of DNCB. At the same time we also observed the effect of MIF antibody pretreatment on the disease active index (DAI) score and the colon pathology by HE staining in the colitis rats. The results obtained showed that the immunofluorescence intensity of double staining of MIF protein in colon nervous tissue and spinal cord was increased in 0.8% and 1.6% DNCB-induced colitis groups than that in the control (60% ethanol) group. Both the colon pathology and the DAI score were significantly reduced by MIF antibody ith pretreatment. Ith injection of 0.8% DNCB after MIF antibody (1:10, 1:5) pretreatment could only induce lower DAI score (P<0.01 as compared, respectively, to the IgG pretreatment group). The colon pathological changes in ith 0.8% DNCB rats were mild, even little after MIF antibody (1:10, 1:5) pretreatment. These results suggest that MIF in spinal cord and enteric nervous system is possibly involved in the rat colitis induced by ith injection of DNCB, which reflects a neuroimmunal mechanism underlying this kind of colitis. MIF is possibly one of the important neurochemical factors in this experimental colitis, even in the UC.
Animals ; Antibodies ; pharmacology ; Colitis, Ulcerative ; chemically induced ; metabolism ; Haptens ; adverse effects ; Injections, Spinal ; Macrophage Migration-Inhibitory Factors ; metabolism ; Rats

OBJECTIVE: To determine the effect of tetramethylpyrazine (TMP) on the expression of migration inhibitory factor (MIF) in acute spinal cord injury (ASCI) in rats. METHODS: Allen's weight-drop method was used to establish a rat model of ASCI at T10. A total of 110 adult SD rats were divided into a sham operation group (group S, n=10), a control group (group C, n=50), and a TMP group (group T, n=50). Spinal cord functionality was measured by a modified Rivilin loxotic plate degree, BBB score, and combined behavioral score (CBS) at 1, 3, 5, 7, 14 and 21 d postoperatively. The injured spinal cord tissue samples were harvested at 1, 3, 6, 12 h and 1, 3, 5, 7, 14, 21 d postoperatively (n=5 at each time point) and used to prepare continuous histological sections, in which the expression of MIF was analyzed by immunohistochemistry. RESULTS: The degree in group T measured by modified Rivlin loxotic plate test after the ASCI was significantly higher than that in group C at 7, 14, and 21 d (P<0.05). BBB score in group T was significantly higher than that in group C at 5, 7, 14, and 21 d after the ASCI (P<0.05). CBS score in group C was significantly higher than that in group T at 5, 7, 14, and 21 d after the ASCI (P<0.05). The significantly low number of MIF positive cells was shown in group T when compared with that in group C at 12 h and 1, 3, 5, 7 d after the ASCI (P<0.05). As time passed, there was negative correlation between modified Rivlin loxotic plate degree and MIF expression and also between BBB score and MIF, and there was positive correlation between CBB score and MIF expression. CONCLUSION TMP has protective effect after the ASCI, and may promote the repair of injured spinal cord tissues. TMP may decrease the MIF expression in cells after the ASCI.
Animals ; Immunohistochemistry ; Intramolecular Oxidoreductases ; metabolism ; Macrophage Migration-Inhibitory Factors ; metabolism ; Pyrazines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; metabolism

OBJECTIVESTo investigate the inhibitory effect of macrophage migration inhibitory factor (MIF) antibody on the proliferation of HepG-2 cells and its mechanism.

METHODSHepG-2 cells were stimulated by different concentrations of MIF antibody (50, 100, 200 and 400 microg/L). The cell survival rates were evaluated by MTT assay. The cell cycles were assessed by flow cytometry (FCM) analysis. Cyclin D1 protein expression was examined by immunohistochemical methods. Vascular endothelial growth factor (VEGF) protein expression was examined by Western blot. ELISA was applied to detect the influence of MIF antibody on the production of IL-6 of HepG-2 cells.

RESULTSHepG-2 cells were inhibited by MIF antibody in a dose and time dependent manner. FCM analysis showed the cell cycles of HepG-2 cells were blocked at G0/G1 phase. With concentrations of MIF antibody of 0, 50, 100, 200, 400 microg/L, the percentages of cells in G0/G1 phase at 48 h were 61.34%+/-1.08%, 65.08%+/-2.71%, 71.19%+/-1.19%, 78.39%+/-1.00%, 83.92%+/-0.51%. With concentrations of MIF antibody of 50, 100, 200, 400 microg/L, the expressions of cyclin D1 protein were 26.06%+/-0.47%, 22.34%+/-0.75%, 18.06%+/-1.16%, 14.03%+/-0.59%, significantly lower than those of the control group (29.51%+/-1.28%). When HepG-2 cells were treated with different concentrations of MIF antibodies of 50, 100, 200, 400 microg/L the expressions of VEGF protein were 21.22%+/-0.68%, 19.64%+/-0.54%, 18.04%+/-0.42%, 16.59%+/-0.66%, significantly lower than those of the control group (23.23%+/-0.51%). With MIF antibody concentrations of 0, 50, 100, 200, 400 microg/L, the exudation amount of IL-6 were correspondingly lower [(210.67+/-9.31) pg/ml, (181.67+/-10.05) pg/ml, (160.50+/-6.60) pg/ml, (143.67+/-10.56) pg/ml, (118.01+/-7.48) pg/m].

CONCLUSIONThe proliferation of HepG-2 cells was inhibited after treatment with MIF antibody. The inhibiting effect is caused by blocking cell cycle progression at G0/G1 phase, decreasing cyclin D1 protein expression, decreasing VEGF protein expression and decreasing the exudation amount of IL-6.

Antibodies ; pharmacology ; Cell Cycle ; Cell Proliferation ; drug effects ; Cyclin D1 ; metabolism ; Hep G2 Cells ; Humans ; Interleukin-6 ; metabolism ; Macrophage Migration-Inhibitory Factors ; immunology ; Vascular Endothelial Growth Factor A ; metabolism

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; blood ; pathology ; Female ; Humans ; Interleukin-8 ; blood ; Liver Neoplasms ; blood ; pathology ; Macrophage Migration-Inhibitory Factors ; blood ; Male ; Middle Aged

Carcinoma, Hepatocellular ; metabolism ; pathology ; Gene Silencing ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Macrophage Migration-Inhibitory Factors ; genetics ; RNA, Small Interfering ; Transfection ; Tumor Cells, Cultured ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo observe the effect of compound shenhua tablet (CST) on the residual kidney expressed macrophage migration inhibition factor (MIF) in rats.

METHODSCST was used to treat 5/6 nephrectomized rats for 12 weeks and the conditions of blood pressure, urinary protein, blood biochemical indices (creatinine, blood urea nitrogen), kidney pathologic change and MIF expression were observed.

RESULTSCST could significantly lower the serum levels of creatinine (P < 0.05), and 24 hrs urinary protein (P < 0.01), reduce the MIF expression and macrophage infiltration in renal glomerulus and tubular mesenchym, and lower the degree of renal glomerular sclerosis and interstitial fibrosis.

CONCLUSIONThe inhibition on the highly expressed MIF may be an important mechanism of the drug in restraining chronic inflammation in residual kidney, delaying the sclerosis and fibrosis progression and protecting renal function.

Albuminuria ; blood ; Animals ; Creatinine ; blood ; Drugs, Chinese Herbal ; pharmacology ; Fibrosis ; pathology ; Kidney ; metabolism ; pathology ; Macrophage Migration-Inhibitory Factors ; metabolism ; Male ; Nephrectomy ; Rats ; Rats, Wistar ; Tablets

OBJECTIVESTo investigate the expression of macrophage migration inhibitory factor (MIF) mRNA in Schwann cells after peripheral nerve injury and roles of Schwann cells and MIF in macrophages activation and nerve regeneration.

METHODSFifty SD rats were divided into 10 groups. One group served as normal control. The rest were anesthetized with 3% sodium pentobarbital (30 - 60 mg/kg, i.p) and sciatic nerves were transected distal to the obturator tendon respectively 1 h, 12 h, 1 d, 3 d, 7 d, 10 d, 14 d, 17 d and 21 d before being killed. Sciatic nerves were resected and connective tissues excised. Schwann cells were obtained by digesting the nerve tissues with trypsin and collagenase. RNA was isolated and reverse-transcription-polymerase chain reaction (RT-PCR) was carried out. cDNA was analyzed by automatic system and the parameters were assessed to define the status of MIF mRNA expression in different groups.

RESULTSThe level of MIF mRNA started to increase 12 h after the nerve transection. The level remained high from day 7 up to 10 after the injury. During the period from days 10 to 21, MIF mRNA decreased slowly to the pre-transection level.

CONCLUSIONAfter peripheral nerve injury, Schwann cells can secrete MIF which may play a pivotal role as an immunomodulatory cytokine in macrophage activation and inflammatory reaction.

Animals ; Female ; Macrophage Migration-Inhibitory Factors ; genetics ; Male ; Peripheral Nerve Injuries ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Schwann Cells ; metabolism