The roles of macrophage migration inhibitory factor in retinal diseases

Macrophage migration inhibitory factor(MIF),a multifunctional cytokine,is secreted by various cells and participates in inflammatory reactions,including innate and adaptive immunity.There are some evidences that MIF is involved in many vitreoretinal diseases.For example,MIF can exacerbate many types of uveitis;measurements of MIF levels can be used to monitor the effectiveness of uveitis treatment.MIF also alleviates trauma-induced and glaucoma-induced optic nerve damage.Furthermore,MIF is critical for retinal/choroidal neovascularization,especially complex neovascularization.MIF exacerbates retinal degeneration;thus,anti-MIF therapy may help to mitigate retinal degeneration.MIF protects uveal melanoma from attacks by natural killer cells.The mechanism underlying the effects of MIF in these diseases has been demonstrated:it binds to cluster of differentiation 74,inhibits the c-Jun N-terminal kinase pathway,and triggers mitogen-activated protein kinases,extracellular signal-regulated kinase-1/2,and the phosphoinositide-3-kinase/Akt pathway.MIF also upregulates Toll-like receptor 4 and activates the nuclear factor kappa-B signaling pathway.This review focuses on the structure and function of MIF and its receptors,including the effects of MIF on uveal inflammation,retinal degeneration,optic neuropathy,retinal/choroidal neovascularization,and uveal melanoma.

Cloning and mRNA expression of macrophage migration inhibitory factor (MIF) gene of large yellow croaker (P seudosciaena crocea)

Mammalian macrophage migration inhibitory factor （MIF） plays an important role as an indispensable mediator in the pathogenesis of inflammatory disease like septicemia, but little is known about the role of MIF homologue in fish septicemia. The authors have cloned the MIF homologue in large yellow croaker Pseudosciaena crocea （LycMIF） using RACE approach. The full-length cDNA of LycMIF was 634 bases and contained an ORF of 345 bases encoding a protein of 115 amino acid residues. As demonstrated by RT-PCR and QRT-PCR assay, MIF mRNAs were constitutively expressed in 11 selected tissues and were abundant in brain and liver. Moreover, the LycMIF transcripts in the liver and head kidney were responsive to bacteria infection and could be significantly up-regulated. Our results provide the first direct evidence that fish MIF was implicated in pathogenesis of fish vibrosis and play an important role in response to bacteria infection.

Brain-derived neurotrophic factor mediates macrophage migration inhibitory factor to protect neurons against oxygen-glucose deprivation

Macrophage migration inhibitory factor(MIF)is a chemokine that plays an essential role in immune system function.Previous studies suggested that MIF protects neurons in ischemic conditions.However,few studies are reported on the role of MIF in neurological recovery after ischemic stroke.The purpose of this study is to identify the molecular mechanism of neuroprotection mediated by MIF.Human neuroblastoma cells were incubated in Dulbecco’s modified Eagle’s medium under oxygen-glucose deprivation(OGD)for 4 hours and then returned to normal aerobic environment for reperfusion(OGD/R).30 ng/mL MIF recombinant(30 ng/mL)or ISO-1(MIF antagonist;50μM)was administered to human neuroblastoma cells.Then cell cultures were assigned to one of four groups:control,OGD/R,OGD/R with MIF,OGD/R with ISO-1.Cell viability was analyzed using WST-1 assay.Expression levels of brain-derived neurotrophic factor(BDNF),microtubule-associated protein 2(MAP2),Caspase-3,Bcl2,and Bax were detected by western blot assay and immunocytochemistry in each group to measure apoptotic activity.WST-1 assay results revealed that compared to the OGD/R group,cell survival rate was significantly higher in the OGD/R with MIF group and lower in the OGD/R with ISO-1 group.Western blot assay and immunocytochemistry results revealed that expression levels of BDNF,Bcl2,and MAP2 were significantly higher,and expression levels of Caspase-3 and Bax were significantly lower in the MIF group than in the OGD/R group.Expression levels of BDNF,Bcl2,and MAP2 were significantly lower,and expression levels of Caspase-3 and Bax were significantly higher in the ISO-1 group than in the OGD/R group.MIF administration promoted neuronal cell survival and induced high expression levels of BDNF,MAP2,and Bcl2(anti-apoptosis)and low expression levels of Caspase-3 and Bax(pro-apoptosis)in an OGD/R model.These results suggest that MIF administration is effective for inducing expression of BDNF and leads to neuroprotection of neuronal cells against hypoxic injury.

Early treadmill exercise increases macrophage migration inhibitory factor expression after cerebral ischemia/reperfusion

The neuroprotective function of macrophage migration inhibitory factor(MIF) in ischemic stroke was rarely evaluated.This study aimed to investigate the effects of early treadmill exercise on recovery from ischemic stroke and to determine whether these effects are associated with the expression levels of MIF and brain-derived neurotrophic factor(BDNF) in the ischemic area.A total of 40 male Sprague-Dawley rats were randomly assigned to the ischemia and exercise group [middle cerebral artery occlusion(MCAO)-Ex,n = 10),ischemia and sedentary group(MCAO-St,n = 10),sham-surgery and exercise group(Sham-Ex,n= 10),or sham-surgery and sedentary group(Sham-St,n = 10).The MCAO-Ex and MCAO-St groups were subjected to MCAO for 60 minutes,whereas the Sham-Ex and Sham-St groups were subjected to an identical operation without MCAO.Rats in the MCAO-Ex and Sham-Ex groups then ran on a treadmill for 30 minutes once a day for 5 consecutive days.After reperfusion,the hanging time tested by the wire hang test was longer and the relative fractional anisotropy determined by MRI was higher in the peri-infarct region of the MCAO-Ex group compared with the MCAO-St group.The expression levels of MIF and BDNF in the peri-infarct region were upregulated in the MCAO-Ex group.Increased MIF and BDNF levels were positively correlated with relative fractional anisotropy changes in the peri-infarct region.There was no significant difference in the levels of MIF and BDNF in the peri-infarct region between the Sham-Ex and Sham-St groups.Our study demonstrated that early exercise(initiated 48 hours after the MCAO) could improve motor and neuronal recovery after ischemic stroke.Furthermore,the increased levels of MIF and BDNF in the peri-infarct region(penumbra) may be one of the mechanisms of enhanced neurological function recovery.All experiments were approved by the Institutional Animal Care and Use Committee in Asan Medical Center in South Korea(2016-12-126).

Macrophage migration inhibitory factor as a potential prognostic factor in gastric cancer

AIM:To investigate macrophage migration inhibitory factor(MIF) expression and its clinical relevance in gastric cancer,and effects of MIF knockdown on proliferation of gastric cancer cells. METHODS:Tissue microarray containing 117 samples of gastric cancer and adjacent non-cancer normal tissues was studied for MIF expression by immunohistochemistry(IHC) semiquantitatively,and the association of MIF expression with clinical parameters was analyzed. MIF expression in gastric cancer cell lines was detected by reverse transcriptionpolymerase chain reaction(RT-PCR) and Western blot. Two pairs of si RNA targeting the MIF gene(MIF si-1 and MIF si-2) and one pair of scrambled si RNA as a negative control(NC) were designed and chemically synthesized. All si RNAs were transiently transfected in AGS cells with OligofectamineTM to knock down the MIF expression,with the NC group and mock group(OligofectamineTM alone) as controls. At 24,48,and 72 h after transfection,MIF m RNA was analyzed by RTPCR,and MIF and proliferating cell nuclear antigen(PCNA) proteins were detected by Western blot.The proliferative rate of AGS cells was assessed by methylthiazolyl tetrazolium(MTT) assay and colony forming assay.RESULTS:The tissue microarray was informative for IHC staining,in which the MIF expression in gastric cancer tissues was higher than that in adjacent noncancer normal tissues(P < 0.001),and high level of MIF was related to poor tumor differentiation,advanced T stage,advanced tumor stage,lymph node metastasis,and poor patient survival(P < 0.05 for all). After si RNA transfection,MIF m RNA was measured by real-time PCR,and MIF protein and PCNA were assessed by Western blot analysis. We found that compared to the NC group and mock group,MIF expression was knocked down successfully in gastric cancer cells,and PCNA expression was downregulated with MIF knockdown as well. The cell counts and the doubling times were assayed by MTT 4 d after transfection,and colonies formed were assayed by colony forming assay 10 d after transfection; all these showed significant changes in gastric cancer cells transfected with specific si RNA compared with the control si RNA and mock groups(P < 0.001 for all).CONCLUSION:MIF could be of prognostic value in gastric cancer and might be a potential target for small-molecule therapy.

Serum and ascites levels of macrophage migration inhibitory factor, TNF-α and IL-6 in patients with chronic virus hepatitis B and hepatitis cirrhosis

Objective: To study the potential role of macrophage migration inhibitory factor (MIF), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the development of chronic virus hepatitis B (CH) and hepatitis cirrhosis (HC). Methods: The serum concentrations of MIF, TNF-α and IL-6 in 18 patients with chronic virus hepatitis B and in 14 patients with hepatitis cirrhosis without as- citic fluid, and the serum and ascites cytokine con- centrations in 22 HC patients with ascitic fluid were detected by enzyme linked immunity sorbed assay. Results: The cytokine concentrations of the patients were significantly higher than those of the controls. The serum levels of MIF, TNF-α and IL-6 of the 22 patients with ascitic fluid were higer than those of 14 HC patients without ascites. In the 18 patients with CH, the serum cytokine concentrations were the low- est. The serum cytokine concentrations of the 22 HC patients with ascites were significantly higher than those of the 14 HC patients without ascites (P< 0. 01). Their serum cytokine concentrations were sig- nificantly higher than those in the 18 patients with CH (P<0. 01). The concentration of IL-6 in ascites was the highest among all the groups. The serum le- vels of MIF, TNF-α and IL-6 are correlated with al- anine aminotransferase (ALT) in the patients with CH, but not in those with HC with or without asci- tes. Conclusions: These results indicated that MIF, TNF- α and IL-6 may participate in the pathological process of CH and cirrhosis, that IL-6 seems to play an important role in ascites formation, and that se- rum levels of MIF, TNF-α and IL-6 appear to reflect the severity of tissue injury in HBV disease.

Upregulation of macrophage migration inhibitory factor and calgizzarin by androgen in TM4 mouse Sertoli cells

Aim： To identify proteins induced by androgen in Sertoli cells during spermatogenesis. Methods： We analyzed protein profiles in TM4 Sertoli cells treated with dihydrotestosterone （DHT） using surface enhanced laser desorption ionization time-of-flight mass spectrometry （SELDI-TOF-MS）. Results： We found increases in the expression of a 5.0-kDa protein at 15 min, an 11.3-kDa protein at 24 h and 4.3 kDa, 5.7 kDa, 5.8 kDa, 9.95 kDa and 9.98 kDa proteins at 48 h after the treatment. In contrast, the expression of 6.3 kDa and 8.6 kDa proteins decreased at 30 min, and 4.9 kDa, 5.0 kDa, 12.4 kDa and 19.8 kDa proteins at 48 h after the treatment. The ll.3-kDa protein was identified as macrophage migration inhibitory factor （MIF） known to having various functions. The 9.98-kDa protein was identified as calgizzarin related to calcium channels. The timing of their expression suggests that MIF and calgizzarin are involved in late regulation of spermatogenesis in Sertoli cells by androgen. Conclusion： MIF and calgizzarin are two important androgen-responsive proteins produced by Sertoli cells and they might play a role in regulating spermatogenesis.

Expression of macrophage migration inhibitory factor in Aspergillus fumigatus keratitis

AIM: To investigate the expression of macrophage migration inhibitory factor(MIF) and detect its role in the innate immune response of fungal keratitis(FK). METHODS: We collected the paraffin-embedded cornea tissues from 10 FK and 6 ocular trauma patients to explore the MIF expression by immunohistochemistry. Then we cultured telomease-immortalized human corneal epithelial cells(THCEs), stimulated by the hyphae suspension of Aspergillus fumigatus(A. fumigatus) to detect the change of MIF with or without the pretreatment of MIF inhibitor [4-Iodo-6-phenylpyrimidine(4-IPP)] by real-time polymerase chain reaction(PCR). The protein level of MIF was also tested by immunohistochemistry, and the level of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) mRNA were compared between normal, hyphae stimulated and 4-IPP pretreated groups by real-time PCR to study the influence of MIF on the expression of TNF-α and IL-6. Corneal severity of rats' FK models was documented by clinical scores, and real-time PCR. Western blot and immunohistochemistry were used to test the expression of MIF, TNF-α and IL-6 in rats' corneas.RESULTS: In the corneas of FK patients, there was much stronger expression of MIF than that in the normal group showed by immunohistochemistry. In cultured THCEs stimulated by A. fumigatus, the expression of MIF became stronger in both immunohistochemistry and PCR at 16, 24, 32 and 48 h post infection(p.i.; P<0.01, P<0.01, P<0.01, P<0.05). After pretreated with 4-IPP, the expression of MIF reduced at 4, 8, 16 h p.i.(P<0.05, P<0.05, P<0.05) and the downstream TNF-α and IL-6 decreased obviously(P<0.05, P<0.01). In rats with A. fumigatus keratitis, the relative mRNA and protein level of MIF increased than thosein the normal group by PCR(at 1 d: P<0.01, 3 d: P<0.01, 5 d: P<0.01), Western blot and immunohistochemistry. After blocked MIF with 4-IPP, the clinical outcomes of rat keratitis showed markedly reduced inflammatory response(P<0.01), with TNF-α and IL-6 decreased in accordance with those in THCEs by PCR(P<0.05, P<0.01). CONCLUSION: The expression of MIF increased significantly in FK patients, THCEs and rats stimulated by A. fumigatus. After blocked with 4-IPP, the expression of MIF reduced, and so did its downstream cytokines: TNF-α and IL-6. The inflammation reaction of the rats' corneas lightened after pretreated with 4-IPP. MIF may play a role in the innate immune response of the corneal resistance against A. fumigatus.

Macrophage migration inhibitory factor regulates proliferation of gastric cancer cells via the PI3K/Akt pathway

AIM:To investigate the effects of macrophage migration inhibitory factor (MIF) on proliferation of human gastric cancer MGC-803 cells and expression of cyclin D1 and p27Kip1 in them,and further determine whether the effects are related to the PI3K/Akt signal transduction pathway. METHODS:Gastric cancer MGC-803 cells were cultured and then treated with 50 μg/L recombinant human MIF (rhMIF) with and without a PI3K inhibitor,LY294002 (25 μmol/L). MTT assay was used to detect the prolifer-ation of MGC-803 cells. Cell cycle was detected by flow cytometry. Expression of cyclin D1 and p27Kip1 mRNA was by reverse transcription-polymerase chain reaction. Protein expression of phosphorylated Akt (p-Akt),Akt,cyclin D1 and p27Kip1 was examined by immunocyto-chemistry and Western blotting. RESULTS:rhMIF signifi cantly stimulated the prolifera-tion of MGC-803 cells and cell cycle progression from G1 phase to S phase in a concentration-and time-de-pendent manner. After the MGC-803 cells were treated with rhMIF for 24 h,the expression of cyclin D1 was signifi cantly up-regulated compared with the cells not treated with rhMIF at both mRNA and protein levels(0.97 ± 0.02 vs 0.74 ± 0.01,P = 0.002; 0.98 ± 0.05 vs 0.69 ± 0.04,P = 0.003). The p27Kip1 was down-regulated but only statistically significant at the protein level. rhMIF significantly increased the expression of p-Akt,which reached the peak at 30 min,but did not affect the expression of Akt. However,LY294002 inhibited all the effects of rhMIF.CONCLUSION:Macrophage MIF increases the proliferation of gastric cancer cells,induces the expression of cyclin D1 at the transcriptional level and inhibits the expression of p27Kip1 at the post-transcriptional level via the PI3K/Akt pathway.

D-dopachrome tautomerase from Japanese sea bass(Lateolabrax japonicus) is a chemokine-like cytokine and functional homolog of macrophage migration inhibitory factor

D-dopachrome tautomerase(DDT),a member of the macrophage migration inhibitory factor(MIF)protein superfamily,is a newly described cytokine with chemokine-like characteristics.However,research on fish DDT remains limited.In this study,we identified a DDT homolog(LjDDT)from the Japanese sea bass,Lateolabrax japonicus.Sequence analysis showed that LjDDT had typical sequence features of known DDT and MIF homologs and was most closely related to DDT of rock bream(Oplegnathus fasciatus).LjDDT transcripts were detected in all tested tissues of healthy Japanese sea bass,with the highest expression found in the liver.Upon infection with Vibrio harveyi,LjDDT transcripts were significantly down-regulated in the three tested tissues,including the liver,spleen,and head kidney.Recombinant LjDDT(rLjDDT)and the corresponding antibody(anti-rLjDDT)were subsequently prepared.The administration of 100μg/g anti-rLjDDT had a statistically significant protective effect on the survival of V.harveyi-infected fish.Moreover,rLjDDT was able to induce the migration of monocytes/macrophages(MO/MФ)and lymphocytes both in vitro and in vivo,but without significant influence on the migration of neutrophils.rLjDDT exhibited chemotactic activity for lipopolysaccharide(LPS)-stimulated M1-type MO/MΦin vitro,but not for cAMP-stimulated M2-type MO/MΦ.Furthermore,the knockdown of LjCD74,but not LjCXCR4,significantly down-regulated the rLjDDT-enhanced migration of MO/MΦand relieved the rLjMIF-inhibited migration of MO/MΦ.These results indicate that LjCD74 may be the major chemotactic receptor of LjDDT and LjMIF in Japanese sea bass MO/MΦ.Combined rLjDDT+rLjMIF treatment had no significant effect on the migration of MsiRNA,LjCD74si-,or LjCXCR4sitreated MO/MΦcompared to the control group,suggesting that the roles of LjDDT and LjMIF may be antagonistic.In conclusion,our study demonstrates for the first time that DDT may play a role in the immune responses of fish against bacterial infection through chemotactic recruitment of MO/MΦvia mediation of CD74 as an antagonist of MIF.

Association of the macrophage migration inhibitory factor promoter polymorphisms with benign lymphoepithelial lesion of lacrimal gland

AIM： To identify the association of the macrophage migration inhibitory factor （MIF） gene polymorphism with the susceptibility of benign lymphoepithelial lesions （BLEL） of the lacrimal gland. METHODS： A total of 40 BLEL of lacrimal gland cases were matched with 40 healthy subjects （HS）. Extraction the plasma and whole blood DNA of patients of lacrimal gland BLEL and HS. Elisa and polymerase chain reaction was used to determine in plasma contents of MIF and MIF gene SNP-173G〉C and STR -794 CATT（8） polymorphism, respectively. RESULTS： The MIF levels in plasma were significantly higher in patients with lacrimal gland BI.EL versus HS （P〈0.001）. The -173 G〉C MIF polymorphism was significantly associated with lacrimal gland BLEL, with a significantly higher frequency of the C allele in lacrimal gland BLEL patients compared with HS （OR=2.38, 95% C1=1.07-5.31, P=0.032）, and the -173 C/x is more frequent in patients than in HS, P=0.037. Besides, we found that the carriage rate of the MIF -173C/x is associated with higher plasma levels of MIF in the BLEI. of lacrimal gland. CONCLUSION： MIF -173G/C variants play an insidious role in susceptibility of BLEL of lacrimal gland. Otherwise,there is no statistically significant correlation exists between MIF-794 CATT （） and BLEL of lacrimal gland.

Macrophage migration inhibitory factor decreased T-type Ca^(2+) channel current through activating Src

Aims T-type Ca2+ current(ICaT)plays an important role in the pathogenesis of atrial fibrillation（AF）.The present study sought to investigate the role of Macrophage migration inhibitory factor（MIF）,a pleiotropic cytokine,in the regulation of T-type Ca2+ channel in atrium myocytes.Methods We used whole-cell voltage-clamp technique and biochemical assays to study the regulation and expression of ICa,T in mouse atrium myocytes（HL-1 cells）.Results Serum MIF concentrations was slightly increased in patients with AF compared to sinus rhythm（SR） controls.In cultured HL-1 cells, significant amounts of MIF were produced in response to hydrogen peroxide（H2O2）,but not AngiotensinⅡ（AngⅡ）. Mouse recombinant MIF（rMIF）（20 or 40 nM,24 h） suppressed peak ICa,T by-38%and-60%in a concentration-dependent manner,impaired the voltage-dependent activation of ICa,T,and down-regulated of TCC alG mRNA.Src inhibitors genistein and PPl significantly enhanced ICaT.The depression of ICa,T induced by rMIF could be reversed by genistein and PP1.Conclusions MIFis involved in the pathogenesis of AF,probably by decreasing ICa,T through impairment of the channel function and activation of c-Src kinases in atrium myocytes.

Effects of antisense oligonucleotides on theexpression of macrophage migration inhibitoryfactor on macrophages

To investigate the effects of antisense oligonucleotides on the expression of macrophage migration inhibitory factor (MIF) on macrophages, the mouse phosphorothioate oligonucleotides were designed and synthesized with the sequences of antisense, 5′-TACGGATACAAGTAGCAC-3′; Sense, 5′-ATGCCTATGTTCATCGTG-3′; Missense, 5′-CTCTCAGACTCGATCTGT-3′. These phosphorothioate oligonucleotides were then transfected into cultured macrophages ( RAW264.7 ) by luciferase vector, and the transfected macrophages were incubated with Lipopolysaccharide (LPS) (1?ng/ml) for various periods of times and collected afterwards. The content of MIF protein in the cultural supernatants was determined by ELISA, cellular RNA extracted and the expression of MIF mRNA was examined by RT-PCR analysis. The experimental results showed that LPS could induce a time-dependent specific expression of MIF on macrophages, in which the MIF mRNA in cells and the MIF protein in cultural supernatants appeared after 3 h and reached their highest concentration at 9-12?h after LPS stimulation. The levels of mRNA and proteins in the macrophages treated with antisense olignucleotides were decreased significantly after stimulation with LPS in comparison with that of stimulation with LPS alone or with that with LPS plus sense or missense oligonucleotides. There were no differences among those without LPS stimulation. It is concluded that macrophages stimulated with LPS express MIF, and the antisense olignucleotides of MIF inhibit the expression of MIF mRNA as well as the secretion of MIF proteins in macrophages.

Moxibustion of Zusanli(ST36)and Shenshu(BL23)alleviates the inflammation of rheumatoid arthritis in rats through regulating macrophage migration inhibitory factor/glucocorticoids signaling

OBJECTIVE:To test the hypothesis that moxibustion may inhibit rheumatoid arthritis(RA)synovial inflammation by regulating the expression of macrophage migration inhibitory factor(MIF)/glucocorticoids(GCs).METHODS:Fifty male Sprague-Dawley rats were randomly divided into five groups(n=10 each):blank Control(CON)group,RA Model(RA)group,Moxibustion(MOX)group,MIF inhibitor(S,R)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid methyl ester(ISO-1)group,and Moxibustion+MIF inhibitor ISO-1(MOX+ISO-1)group.Rats in the ISO-1 group and ISO-1+MOX group were intraperitoneally injected with the inhibitor ISO-1.The rats in the RA group,ISO-1 group,MOX group,and ISO-1+MOX group were injected with Freund's complete adjuvant(FCA)in the right hind footpad to establish an experimental RA rat model.In the MOX group and MOX+ISO-1 group,rats were treated with Moxa.The thickness of the footpads of the rats in each group was measured at three-time points before,after modeling and after moxibustion treatment.The contents of serum MIF,corticosterone(CORT),tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)were detected by enzyme-linked immunosorbent assay;and the contents of synovial MIF were detected by Western blot.Hematoxylin-eosin(HE)staining method was used to observe the pathological changes of synovial tissue under a section light microscope,and pathological scoring was performed according to the grading standard of the degree of synovial tissue disease.RESULTS:Moxibustion was found to reduce the level of MIF and alleviate inflammation in RA rats in this study.In addition,after inhibiting the expression of MIF,the level of CORT increased,and the level of TNF-α decreased.Treating RA rats with inhibited MIF by moxibustion,the level of CORT was almost unchanged,but the level of TNF-α further decreased.The correlation analysis data suggested that MIF was positively related to the expression of TNF-α and negatively correlated with the expression of CORT.CONCLUSION:Reducing MIF to increase CORT and decrease TNF-α by moxibustion treatment in RA.MIF may be a factor for moxibustion to regulate the expression of CORT,but the expression of TNF-α is due to the incomplete regulation of the MIF.This study added to the body of evidence pointing to moxibustion's antiinflammatory mechanism in the treatment of RA.

血清MIF、MCP-1、suPAR水平与脓毒症严重程度及合并ARDS风险的关系

目的探讨血清巨噬细胞迁移抑制因子(MIF)、单核细胞趋化蛋白-1(MCP-1)、可溶性尿激酶型纤溶酶原激活物受体(suPAR)水平与脓毒症严重程度及合并急性呼吸窘迫综合征(ARDS)风险的关系。方法回顾性分析2022年2月至2023年5月太原钢铁(集团)有限公司总医院收治的86例脓毒症患者的临床资料。依据病情程度不同将患者分为脓毒症组(n=20)、严重脓毒症组(n=48)和脓毒症休克组(n=18)。入院72 h内参考ARDS诊断标准将患者分为ARDS组(n=27)和非ARDS组(n=59)。检测并比较各组脓毒症患者血清MIF、MCP-1、suPAR水平。收集ARDS组与非ARDS组患者年龄、性别、体重指数、合并症、感染类型、既往史、心率、急性生理学和慢性健康状况评价Ⅱ(APACHEⅡ)、脓毒症相关性器官衰竭评价(SOFA)评分、白细胞计数、血乳酸、天冬氨酸转移酶(AST)、丙氨酸转移酶(ALT)、总胆固醇等指标。采用多因素Logistic回归分析对影响脓毒症患者并发ARDS的危险因素进行分析。通过受试者工作特征(ROC)曲线分析血清MIF、MCP-1、suPAR水平预测脓毒症患者并发ARDS的价值。结果脓毒症休克组患者血清MIF、MCP-1、suPAR水平分别为(94.02±10.13)、(506.55±45.15)、(13.89±3.95)ng/mL,均高于脓毒症组[(76.93±7.01)、(148.38±35.74)、(6.07±2.13)ng/mL]和严重脓毒症组[(85.46±8.74)、(327.08±40.62)、(8.42±1.07)ng/mL],而严重脓毒症组患者血清MIF、MCP-1、suPAR水平均高于脓毒症组,差异均有统计学意义(P<0.05)。ARDS组与非ARDS组患者的年龄、性别构成比、体重指数、合并症、感染类型、白细胞计数、心率、吸烟史、饮酒史、血乳酸、AST、ALT、总胆固醇比较,差异均无统计学意义(P>0.05);ARDS组患者APACHEⅡ评分、SOFA评分、有急腹症和胰腺炎占比及血清MIF、MCP-1、suPAR水平均高于非ARDS组,差异均有统计学意义(P<0.05)。经多因素Logistic回归分析结果显示,急腹症、胰腺炎、APACHEⅡ评分、SOFA评分、MIF、MCP-1、suPAR是影响脓毒症患者并发ARDS的独立危险因素(P<0.05)。经ROC曲线分析结果显示,血清MIF、MCP-1、suPAR水平均能预测脓毒症患者ARDS的发生,曲线下面积分别为0.904、0.910、0.917,预测价值较好(P<0.05)。结论血清MIF、MCP-1、suPAR水平与脓毒症患者病情程度、并发ARDS密切相关,且血清MIF、MCP-1、suPAR水平对ARDS的发生有较好的预测价值。

sVCAM1、MIF及IgE在支气管哮喘急性发作患者血清中的表达水平及其意义

【目的】探讨可溶性血管内皮细胞表达的粘附因子(sVCAM1)、巨噬细胞迁移抑制因子(MIF)和免疫球蛋白E(IgE)在支气管哮喘急性发作期患者血清中的表达水平及其意义。【方法】选取2018年10月至2021年3月渭南市妇幼保健院收治的126例支气管哮喘急性发作患者(急性发作组)和110例支气管哮喘非急性发作患者(非急性发作组),另选择100例体检健康志愿者作为对照组,比较三组血清sVCAM1、MIF及IgE水平。统计支气管哮喘急性发作患者住院期间预后情况分为预后不良组和预后良好组,比较预后不良组和预后良好组的临床资料。Logistic多因素回归分析支气管哮喘急性发作患者预后的影响因素;采用受试者工作特征(ROC)曲线分析血清sVCAM1、MIF及IgE对支气管哮喘急性发作患者预后的预测价值。【结果】急性发作组、非急性发作组血清sVCAM1、MIF及IgE水平均高于对照组(P<0.05),且急性发作组患者血清sVCAM1、MIF及IgE水平高于非急性发作组(P<0.05)。本研究中支气管哮喘急性发作患者住院期间不良预后发生率为16.67%。预后不良组住院时间、病情为重度占比及sVCAM1、MIF、IgE水平均高于预后良好组(P<0.05),一秒用力呼气容积(FEV 1)/用力肺活量(FVC)低于预后良好组(P<0.05)。Logistic多因素分析结果显示:病情为重度及血清sVCAM1升高、MIF升高、IgE升高均是影响支气管哮喘急性发作患者预后的危险因素(OR=2.795、3.142、3.022、3.449,P<0.05)。ROC曲线分析结果显示:血清sVCAM1、MIF及IgE预测支气管哮喘急性发作患者预后的最佳截断点分别为467.92 ng/L、51.02 ng/mL、285.96 IU/mL,三者联合预测支气管哮喘急性发作患者预后的曲线下面积(AUC)为0.890,高于血清sVCAM1、MIF及IgE单独预测的AUC(P<0.05)。【结论】血清sVCAM1、MIF及IgE在支气管哮喘急性发作期患者血清中的表达水平异常升高,且三者联合预测支气管哮喘急性发作期患者预后的效能较高。

Design of Agents Interacting with Immunoregulating Proteins:Potential Inhibitors of the Phenylpyruvate Tautomerase Activity Catalysed by Macrophage Migration Inhibitory Factor (MIF)

The macrophage migration inhibitory factor has been implicated in a number ofimmune and inflammatory processes. MIF presents particular opportunities for drug design anddevelopment with potential therapeutic applications. Drug design strategies taking intoconsideration of specific stereochemical and tautomeric requirements in the interaction of MIF withsubstrates and inhibitors allow several novel structures to be designed. Our investigationssuccessfully explored the tautomeric and stereochemical aspects of new compounds of the2-phenylpyruvic acid type, both experimentally, through synthesis and structural investigations andcomputationally, through molecular mechanics and quantum mechanics calculations.

MIF通过调节急性幽门螺杆菌感染介导的炎症反应、自噬和凋亡促进胃癌细胞增殖及细胞周期进展

目的:探究巨噬细胞迁移抑制因子(MIF)在幽门螺杆菌(Hp)感染的胃癌细胞中的表达及MIF对Hp感染的胃癌细胞增殖和凋亡的影响及相关机制。方法:体外培养胃癌细胞系AGS,Western blot检测随Hp感染时间的延长,AGS细胞中MIF蛋白表达变化;siRNA干扰与脂质体瞬时转染技术靶向沉默胃癌细胞中MIF表达,RT-PCR与Western blot检测si-MIF转染效率;Hp感染转染后的胃癌细胞,并将其分为:si-NC组、si-MIF组、Hp+si-NC组、Hp+si-MIF组,MTT检测各组胃癌细胞增殖活力,Annexin V-FITC/PI双染色联合流式细胞术检测各组细胞凋亡率,DCFH-DA法检测各组细胞中活性氧(ROS)含量,JC-1染色观察各组细胞线粒体膜电位水平,Western blot检测各组细胞线粒体自噬信号通路PINK1/Parkin中蛋白PINK1、Parkin和自噬标志蛋白LC3Ⅱ/LC3Ⅰ、p62表达,免疫荧光检测各组胃癌细胞中自噬蛋白LC3B与线粒体特征蛋白Mito Tracker的共定位水平。结果:胃癌细胞中MIF蛋白水平随Hp刺激时间延长而逐渐升高,且与0 h相比,Hp感染12 h时MIF升高最为显著(P<0.05);与转染si-NC相比,si-MIF能显著下调胃癌细胞中MIF mRNA与蛋白表达(P<0.05);与si-NC组相比,Hp+si-NC组和Hp+si-MIF组胃癌细胞增殖能力与凋亡率均显著升高(P<0.05),si-MIF组增殖能力明显降低(P<0.05),但凋亡率无明显变化(P>0.05),而Hp+si-MIF组增殖能力较Hp+si-NC组显著降低(P<0.05),但凋亡率明显升高(P<0.05);DCFH-DA与JC-1染色检测结果显示,与si-NC组相比,Hp+si-NC组与Hp+si-MIF组ROS显著增加,线粒体膜电位明显降低(P<0.05),si-MIF组无明显变化(P>0.05),Hp+si-MIF组ROS水平与线粒体膜电位水平变化趋势较Hp+si-NC组更为明显(P<0.05)。Western blot与免疫荧光染色检测结果显示,与si-NC组相比,Hp+si-NC与Hp+si-MIF组的LC3Ⅱ/LC3Ⅰ、PINK1、Parkin蛋白表达及与LC3B共定位的线粒体数均显著升高(P<0.05),p62蛋白表达明显降低(P<0.05),si-MIF组均无明显变化(P>0.05);与Hp+si-NC组相比,Hp+si-MIF组的上述检测指标变化均显著降低(P<0.05)。结论:Hp感染能促进胃癌细胞中MIF表达,并通过上调MIF介导的PINK1/Parkin线粒体自噬水平起到保护线粒体功能的作用,从而促进肿瘤细胞增殖,抑制Hp诱导的细胞凋亡,而沉默胃癌细胞中MIF表达,能显著削弱Hp的上述作用。

甲状腺乳头状癌患者术后血清MMIF和IL-6表达水平与甲状旁腺功能减退持续时间的相关性研究

目的探究甲状腺乳头状癌患者术后血清巨噬细胞移动抑制因子(macrophage movement inhibitory factor,MMIF)和白细胞介素(interleukin,IL)-6表达水平与甲状旁腺功能减退持续时间的关系。方法选取北京市平谷区医院2020年5月~2022年2月133例甲状腺乳头状癌术后甲状旁腺功能减退患者,根据甲状旁腺功能减退持续时间的中位数将其分为长时组(>8周,n=65)和短时组(≤8周,n=68)。酶联免疫吸附法检测血清MMIF和IL-6水平,并分析二者与甲状旁腺功能减退持续时间的关系。结果长时组和短时组的年龄、女性占比、体质量指数、肿瘤直径、双侧中央区淋巴结清扫占比、术后引流量、包膜侵犯占比、甲状旁腺意外切除占比、术前血钙和甲状旁腺激素、术后第1天的肿瘤坏死因子-α水平,差异均无统计学意义(t/χ^(2)/Z=0.123~1.401,均P>0.05)。与短时组相比,长时组的手术时间[114(102,139)min vs 101(95,114)min]、术后第1天的MMIF(2.38±0.26 ng/ml vs 2.09±0.19ng/ml)和IL-6水平[14.10(12.47,15.13)ng/L vs 12.42(11.61,13.38)ng/L]均升高,差异具有统计学意义(Z/t=3.419~7.185,均P<0.05)。Pearson分析结果显示log10(甲状旁腺功能减退持续时间)与MMIF呈正相关关系(r=0.397,P=0.000)。Spearman分析结果显示甲状旁腺功能减退持续时间与手术时间和IL-6均呈正相关关系(rs=0.363,P=0.000;rs=0.281,P=0.001)。线性回归分析结果显示手术时间(t=2.734,P=0.007)、MMIF(t=4.265,P=0.000)和IL-6(t=3.006,P=0.003)均与甲状旁腺功能减退持续时间独立相关。结论MMIF和IL-6与甲状旁腺功能减退持续时间呈正相关关系,二者水平高提示患者甲状旁腺功能减退持续时间长。

慢性牙周炎患者龈沟液MIF、β-catenin表达与疾病严重程度的关系

目的探讨慢性牙周炎患者龈沟液巨噬细胞移动抑制因子(MIF)、β-连环蛋白(β-catenin)表达与疾病严重程度的关系。方法选取2019年1月至2021年1月丽水市人民医院口腔科就诊的慢性牙周炎患者80例,其中轻中度牙周炎组36例、重度牙周炎组44例。另选取同期就诊的40名牙周组织健康者作为对照组。采用ELISA法检测龈沟液MIF表达水平,采用RT-qPCR法检测龈沟液β-catenin表达水平。比较3组龈沟液MIF、β-catenin表达水平。分析龈沟液MIF、β-catenin表达水平与慢性牙周炎疾病严重程度的相关性及对慢性牙周炎的诊断价值。结果重度牙周炎组、轻中度牙周炎组、对照组龈沟液MIF表达水平分别为(14.63±1.95)、(12.38±1.74)、(10.16±1.61)ng/ml,β-catenin表达水平分别为(3.18±0.98)、(1.85±0.57)、(1.29±0.12)g/L。重度牙周炎组龈沟液MIF、β-catenin表达水平>轻中度牙周炎组>对照组(均P<0.05)。Spearman等级相关性分析显示,龈沟液MIF、β-catenin表达水平与慢性牙周炎疾病严重程度均呈正相关(rs=0.744、0.808,均P<0.05)。ROC曲线分析显示,龈沟液MIF、β-catenin表达水平诊断慢性牙周炎的最佳截断值分别为11.930 ng/ml、1.500 g/L,灵敏度均为0.900,特异度分别为0.950、0.990,均有较高的诊断价值(均P<0.05)。结论慢性牙周炎患者龈沟液MIF、β-catenin表达水平均升高,且与疾病严重程度呈正相关。该两项指标可用于评估疾病严重程度,且有较高的诊断价值,或可为治疗方案的选择提供参考。