The interaction between Cyclophilin A and CD147 and its clinical significance in periodontal diseases / 口腔疾病防治

@#Cyclophilin A (CypA) is the first foldable enzyme in human cells that has peptidyl proliferase-trans isomerase activity and has a strong proinflammatory effect. CD147 can act as the signal receptor of CypA. The interaction of the two through cell-surface heparin binding activates extracellular regulated protein kinases (ERK1/2) and nuclear factor kappa-B (NF-κB) signaling pathways in macrophages and increases the expression of MMPs and other inflammatory factors. The CypA/CD147 interaction regulates inflammation, promotes the inflammatory response and bone resorption and is involved in the pathological processes of a variety of systemic diseases. CypA and CD147 may take part in the chemotaxis of inflammatory cells, increase white blood cell infiltration in tissues, and increase CypA and CD147 expression in periodontitis gum tissue and gingival groove liquid with inflammation, prompting their interaction to promote the progression of periodontitis. However, the specific function of the signaling pathways in the periodontitis mechanism still requires further elucidation.

Expression of cyclophilin A in oral squamous cell carcinoma and its effect on cell proliferation and invasion / 华西口腔医学杂志

OBJECTIVES@#To investigate the expression of cyclophilin A (CyPA) in oral squamous cell carcinoma (OSCC) and explore the effect of downregulating the expression of CyPA gene on the proliferation and invasion of SCC-25 cells.@*METHODS@#A total of 77 cases of patients with OSCC were selected. The expression levels of CyPA proteins in OSCC and adjacent normal tissues were evaluated. SCC-25 cells were cultured and divided into the CyPA interference sequence group, negative control group, and blank group. The expression levels of CyPA mRNA and protein in cells were detected by using real-time fluorescent quantitative polymerase chain reaction and Western blot, respectively. Cell proliferation was detected by using methyl thiazolyl tetrazolium and plate colony formation assays. Cell invasion was detected by using Transwell assay.@*RESULTS@#The positive expression rate of CyPA protein in OSCC tissues was 76.62%, which was higher than that in adjacent tissues (@*CONCLUSIONS@#The CyPA protein is highly expressed in OSCC tissues, and the downregulation of CyPA gene expression in SCC-25 cells can reduce cell proliferation and inhibit cell invasion.

Role of cyclophilin A during coronavirus replication and the antiviral activities of its inhibitors / 生物工程学报

Cyclophilin A (CypA) is a widely distributed and highly conserved protein in organisms. It has peptidyl-prolyl cis/trans isomerase activity and is a receptor for cyclosporin A (CsA). Coronaviruses are enveloped, single-stranded, positive-sense RNA viruses. Seven types of coronaviruses are currently known to infect humans, among which SARS-CoV, MERS-CoV, and SARS-CoV-2 are fatal for humans. It is well established that CypA is essential for the replication of various coronaviruses such as SARS-CoV, CoV-229E, CoV-NL63, and FCoV. Additionally, CsA and its derivatives (ALV, NIM811, etc.) have obvious inhibitory effects on a variety of coronaviruses. These results suggest that CypA is a potential antiviral target and the existing drug CsA might be used as an anti-coronavirus drug. At the end of 2019, SARS-CoV-2 raged in China, which seriously theatern human health and causes huge economic lases. In view of this, we describe the effects of CypA on the replication of coronaviruses and the antiviral activities of its inhibitors, which will provide the scientific basis and ideas for the development of antiviral drugs for SARS-CoV-2.

Mass Spectrometric Analysis of Proteoforms of Cyclophilin A from Human Colorectal Carcinoma Tissues / 分析化学

Cyclophilin A was isolated from human colorectal carcinoma tissues by ultrafiltration,affinity chromatography,and two dimensional electrophoresis.By the aid of Western blot analysis,several isoforms of cyclophilin A were detected.,The tryptic digests from cyclophilin A were analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry as well as electrospray tandem mass spectrometry techniques.Several different modifications such as N-terminal acetylation (Ac-1Met and Ac-2Val) and phosphorylation of 5Thr were identified from these isoforms of cyclophilin A,which represented the majority of the proteoforms of cyclophilin A in this study.In addition,other kinds of modifications such as oxidation and deamidation were also found in these proteoforms.The results indicated that the developed method could be used to rapidly and correctly identify the several proteoforms of cyclophilin A isolated from human colorectal carcinoma tissues.

Expression of cyclophilin A in foaming of macrophages induced by silica / 中国职业医学

OBJECTIVE: To investigate the role of cyclophilin A in the foaming process of macrophages induced by free silica( SiO_2). METHODS: The human peripheral blood mononuclear cell THP-1 in the logarithmic growth phase was induced and differentiated into human macrophages with phorbol 12-myristate 13-acetate at 100 μg/L for 48 hours. The cells were divided into 4 groups. The cells in the blank control group were not treated. The cells in the oxidized low-density lipoprotein( ox-LDL) control group were treated with a final concentration of 50 mg/L ox-LDL. The cells of 50 mg/L SiO_2 group were treated with 50 mg/L of SiO_2 and ox-LDL. The cells of 100 mg/L SiO_2 group were treated with 100 mg/L of SiO_2 and 50 mg/L of ox-LDL. After treatment of cells for 48 h,cell viability was measured by MTS method. The lipid accumulation of cells was observed by oil red O staining and colorimetric method; the expression of cyclophilin A in cell supernatant was detected by enzyme-linked immunosorbent assay,and the expression of cyclophilin A in the cells was detected by Western blotting. RESULTS: The cell viability was the highest when the concentration of SiO_2 was 100 mg/L compared with the control and other four SiO_2groups( P < 0. 01). The cell foaming change in the 100 mg/L SiO_2 group observed by oil red O staining significantly increased compared with the blank control group. The expression of total cholesterol,free cholesterol increased in the ox-LDL control group,50 mg/L SiO_2 group and 100 mg/L SiO_2group( P <0. 05),and the cholesterol specific gravity increased( P < 0. 05) compared with the blank control group,meanwhile the expression of cyclophilin A in the cell supernatant increased( P < 0. 05),and the expression in the macrophages cells decreased( P < 0. 05). CONCLUSION: Cyclophilin A is involved in the foaming process of macrophages induced by SiO_2.

Effect of extracellular cyclophilin A on inflammatory response and anti-inflammatory activity of antibody against cyclophilin A / 生物工程学报

Cyclophilin A (CypA) is a member of peptidyl prolylisomerases (PPIase) family. CypA is best known as a ubiquitously distributed intracellular protein. It has also been shown to be secreted by cells in response to inflammatory stimuli and oxidative stress. Extracellular CypA (eCypA) interacts with CD147 to initiate inflammatory responses via recruiting leucocytes into inflamed tissue. Recombinant CypA was expressed in Escherichia coli and then purified using Superdex 75™ 16/60. The results of Real-time PCR and ELISA showed that the expression levels of proinflammatory cytokines, such as IL-1β, secreted by eCypA stimulated BMDM were significantly up-regulated, indicating that eCypA played an important role in promoting inflammatory responses. In addition, anti-CypA antibody was prepared using purified CypA protein for therapeutic evaluation in a mouse model of LPS-induced acute lung inflammation. Antibody-treated mice showed reduced lung injury and the expression levels of IL-1β in the lung tissue and blood were decreased significantly, indicating that anti-CypA antibody exerted a potent anti-inflammatory activity. Our findings provide a potential therapeutic antibody for inflammation-mediated diseases.

Selective targeting p53WT lung cancer cells harboring homozygous p53 Arg72 by an inhibitor of CypA / 中国药理学与毒理学杂志

OBJECTIVE To explored the potential of pharmacological stabilization and reactivation of p53 for targeted cancer therapies. METHODS The cytotoxicity of a potent Cyclophilin A (CypA) inhibitor HL001 was tasted against a panel of cancer cell lines. The genotypes and activation of p53 were compared with the cytotoxicity profile of HL001. Two-dimensional (2D) PAGE analysis was performed to investigate differentially expressed proteins that involves in the anti-proliferation effects of HL001. Pull-down and Co-IP were used to confirmed the new identified PPI between CypA and G3BP1 and orthotopic animal model of lung cancer was used to tested the anti- tumor activity of HL001 in vivo. RESULTS We identify a novel CypA small molecule inhibitor HL001 that induces non-small cell lung cancer (NSCLC) cell cycle arrest and apoptosis via restoring p53 expression. We find that HL001 stabilizes p53 through inhibiting the MDM2-mediated p53 ubiquitination. Further mechanistic studies reveal that the downregulation of G3BP1 and the induction of reactive oxygen species and DNA damage by HL001 contribute to p53 stabilization. Surprisingly, HL001 selectively suppresses tumor growth in p53 wildtype NSCLC harboring Arg72 homozygous alleles (p53- 72R) through disrupting interaction between MDM2 and p53-72R in a CypA dependent manner. Moreover, combining HL001 with cisplatin synergistically enhance tumor regression in orthotopic NSCLC mouse model. CONCLUSION Pharma?cologic inhibition of CypA offers a potential therapeutic strategy via specific activation of p53-72R in NSCLC.

Effects of Tongxinluo Capsules on Serum CyPA and MMP-9 Levels in Patients with Coronary Heart Disease after PCI / 中国药房

OBJECTIVE:To investigate the effects of Tongxinluo capsules on the levels of serum cyclophilin A (CyPA) and matrix metalloproteinase-9 (MMP-9) in patients with coronary heart disease (CHD) after percutaneous coronary intervention (PCI).METHODS:A total of 115 CHD patients undergoing PCI were randomly divided into control group (59 cases) and observation group (56 cases).Control group was given conventional treatment.Observation group was additionally given Tongxinluo capsules 0.78 g,3 times a day,for consecutive 6 months,on the basis of control group.The levels of CyPA and MMP-9 as well as the occurrence of ADR were observed in 2 groups before and after treatment.RESULTS:Before treatment,there was no statistical significance in serum level of CyPA and MMP-9 between 2 groups (P＞0.05).The levels of CyPA in 2 groups 1 d and 1 month after surgery as well as the level of CyPA in control group 6 months after surgery were all significantly higher than before surgery,but the observation group was significantly lower than the control group 1 and 6 months,this index was decreased gradually as time,with statistical significance (P＜0.05).There was no statistical significance in the levels of CyPA between 2 groups 1 d after surgery or in observation group between 6 months after surgery and before surgery (P＞0.05).The levels of MMP-9 in 2 groups 1 d and 1 month after surgery were significantly higher than before surgery,and the observation group was significantly lower than the control group,this index was decreased gradually as time,with statistical significance (P＜0.05).There was no statistical significance in the levels of MMP-9 of 2 groups between 6 months after surgery and before surgery(P＞0.05).There was no statistical significance in the incidence of ADR between 2 groups (P＞0.05).CONCLUSIONS:Based on routine treatment,Tongxinluo capsules can significantly reduce the levels of CyPA and MMP-9 in CHD patients after PCI.

Endothelial-dependent Vasodilatation and Expressions of CyPA, p-ERK1/2 in Experimental Rats With Obesity Combining Atherosclerosis / 中国循环杂志

Objective: To observe the endothelial-dependent vasodilatation and expressions of cyclophilin A (CyPA), phosphorylated extracellular signal regulated kinase1/2 (p-ERK1/2) in experimental rats with obesity combining atherosclerosis. Methods: A total of 30 male Wistar rats were randomly divided into 3 groups:Control group, the rats received basic diet followed by intraperitoneal injection of normal saline;Atherosclerosis (AS) group, the rats received basic diet for 8 weeks followed by high cholesterol diet with intraperitoneal injection of a single dose vitamin D3 600,000 IU/kg; Obesity+AS group, the rats received high cholesterol diet for 8 weeks (which made their body weights at 20%higher than the other 2 groups) followed by intraperitoneal injection of a single dose vitamin D3 600,000 IU/kg. n=10 in each group. 16 weeks later, the endothelial-dependent vasodilatation was examined in all rats, expressions of CyPA and p-ERK1/2 in arterial wall were detected by HE staining and immunohistochemistry. Results: Compared with Control group, both AS group and Obesity+AS group had reduced endothelial-dependent vasodilatation (72.49 ± 3.27)%and (42.28 ± 2.62)%vs (96.63 ± 3.85)%, such reduction was even more in Obesity+AS group (42.28 ± 2.62)%vs (72.49 ± 3.27)%, all P Conclusion: The rats with obesity and AS had decreased endothelial-dependent vasodilatation, severe atherosclerosis and calciifcation plaques, increased expressions of CyPA and p-ERK1/2, which speculated that obesity might be an independent risk factor for atherosclerosis.

Relationship Between Blood Levels of Cyclophilin A and Chronic Heart Failure / 中国循环杂志

Objective: To explore the relationship between blood levels of cyclophilin A (CyPA) and chronic heart failure (CHF). Methods: A total of 166 CHF patients were enrolled as CHF group, according to NYHA classiifcation, it was further divided into 4 sub-groups: Class I,n=37, Class II,n=39, Class III,n=46 and Class IV,n=44. In addition, there was a Normal control group,n=52. Blood levels of CyPA, B-type natriuretic peptide (BNP) and high sensitivity C-reactive protein (hs-CRP) were examined and compared among different groups. Results: Compared with Normal control group, CHF group had elevated CyPA (5.11 ± 2.43) ng/ml vs (2.28 ± 0.61) ng/ml, BNP (385.65 ± 184.06) pg/ml vs (90.37 ± 18.44) pg/ml and hs-CRP (11.74 ± 5.44) mg/L vs (5.99 ± 1.14) mg/L, all P0.05. Pearson correlation analysis indicated that blood levels of CyPA were positively related to BNP and hs-CRP in CHF patients (r=0.838,P Conclusion: Blood levels of CyPA were elevated in CHF patients and it’s obviously related to NYHA classiifcation, which might have certain effects on CHF diagnosis and evaluation.

Mechanism of Cyclophilin A in tumor / 国际肿瘤学杂志

Cyclophilin A (CypA) is found to be highly expressed in different kinds of tumor cells,which could regulate the occurrence and development of many kinds of tumor through multiple signal transduction pathways such as inducing the formation of inflammatory carcinoma,accelerating the transcription cycle of tumor cells,promoting the invasion and metastasis of tumor cells,inhibiting the apoptosis of tumor cells and reducing the sensitivity of tumor cells to chemotherapy drugs.It suggests that CypA might be considered as a kind of oncogene,which is expected to be a novel target for tumor treatment.

Expression of cyclophilin A in gastric cancer and its relationship with proliferation of gastric cancer cells / 第二军医大学学报

Objective To examine the expression of cyclophilin A (CypA) in gastric cancer tissues and cell lines, and to explore the effect and mechanism of CypA on the proliferation of gastric cancer cells. Methods Real-time quantitative PCR and Western blotting analysis were used to detect the expression of CypA in gastric cancer tissues, the corresponding adjacent tissues and gastric cancer cell lines. Small interfering RNAs targeting CypA were synthesized and transfected into the gastric cancer cell line MKN45. The effect of CypA-siRNA on endogenous CypA expression was observed; meanwhile, nonspecific siRNA control group and negative control group were also designed. Cell proliferation was measured with MTT assay in each group; cell cycle was detected with flow cytometry. The expression of PCNA, P21, P16, and CyclinD1 gene, involved in cell proliferation, were also detected by real-time quantitative PCR and Western blotting analysis. Results CypA expression was significantly up-regulated in gastric cancer tissues compared with para-cancer tissues; and CypA expression in gastric cancer cell lines was also significantly higher than that in the gastric epithelial cell line GES-1, with the highest expression found in the poorly-differentiated gastric cancer cell line MKN45(P<0. 05). The expression of CypA was significantly inhibited by the CypA-siRNA in MKN45 (P<0. 05). MTT assay showed that the inhibition of CypA by CypA-siRNA significantly suppressed MKN45 cell proliferation (P<0. 05). The ratio of G0/G1 phase cells was significantly increased in MKN45 cells after CypA-siRNA transfection, while the ratio of G2/M phase was significantly decreased (P<0. 05). In addition, the expression of PCNA, Cyclin D1 was significantly decreased at mRNA and protein levels when CypA was inhibited, and the expression of P21 was significantly increased (P<0. 05). Conclusion CypA is overexpressed in gastric cancer cells; CypA gene can effectively promote gastric cancer cell proliferation by regulating some proliferation related genes.

Study on the Relationship between Cyclophilin A and Chronic Heart Failure / 现代检验医学杂志

Objective To investigate the diagnostic value of cyclophilin A(CyPA)for chronic heart failure (CHF).Methods Eighty pateints,made a definite diagnosis as CHF,were classified 30 cases as A phase,29 cases as C phase,21 cases as D phase of them.30 healthy subjects were enrolled in this study.Serum CyP A was detcted by ELISA.NT-proBNP was detec-ted with Roche Cobas 6 000 automatic electrochemilu-minescence immunoassay system.Results The amounts of serum CyP A(x-±s,ng/ml)in healthy subjects and each group with CHF were 110.10±49.73,327.85±82.67,331.70±69.34 and 342.46±92.55.Using Oneway Anova,CyP A concentration of CHF was significantly higher than healthy controls (F=58.45,P0.05).Results of ROC curves showed the AUC (CyP A)was 0.97.The best cutoff value was 198.39 (ng/ml).The sensitivity was 91.30%,the specificity was 93.30%.The correlation analysis showed that CyP A and NT-proBNP were correlated (r=0.30,P<0.01). CyP A and NT-proBNP combined detections showed the sensitivity was 93.80% and the specificity was 100%.Conclusion The Cyp A levels in CHF group are significantly higher than the control group,suggesting that Cyp A may be a factor for CHF appearance.These results indicate that combined detections may helpful to early diagnosis of CHF.

The Changes of Endothelial-dependent Vasodilatation and Cyclophilin A Expression at Different Phases of Atherosclerosis in Experimental Rats / 中国循环杂志

Objective: To observe the changes of endothelial-dependent vasodilatation and cyclophilin A (CyPA) expression at different phases of atherosclerosis in experimental rats. Methods: A total of 30 male Wistar rats were divided into 4 groups: Control group, the rats received normal diet, and 3 atherosclerosis groups, the rats received high cholesterol diet for different period of time and a single dose intraperitoneal injection of Vitamin D3 at 600,000 IU/kg, as Atherosclerosis at 8 weeks (AS 8W) group, AS 12W group and AS 15W group, n=8 in each AS group. The rats were sacriifced at the same time to isolate aorta abdominalis. The expression of CyPA at the wall of aorta abdominalis was detected by HE staining and immunohistochemistry. In addition, the aorta abdominalises was cut into 5 mm rings to observe its response to acethylcholine in exvivo organ bath. Results: With prolonged modeling time, the rats at different groups presented normal vessel structure, endothelial cell damage, vessel smooth muscle cell proliferation, atherosclerosis plaque formation and calciifed plaque formation for differentpathological characteristics. The endothelial-dependent vasodilatation was decreased and the maximum vasodilatation percentage in Control group, AS 8W group, AS 12W group and AS 15W group were at (93.46 ± 2.80) %, (82.58 ± 3.25) %, (61.19 ± 3.72)% and (41.28 ± 2.68)% respectively,P<0.05 between each group. The CyPA expression in endothelial cell and vessel smooth muscle cell were increased accordingly in 4 groups by OD value as (0.25 ± 0.06), (0.34 ± 0.09), (0.53 ± 0.09) and (0.68 ± 0.13) respectively,P<0.05 between each group. Conclusion: The CyPA expression increased and the endothelial-dependent vasodilatation decreased with the progress of atherosclerosis accordingly in experimental rats. The expression level of CyPA is related to atherosclerosis degree and it is one of the initial factors for atherosclerosis in rats.

Effects of CyPA signal pathway in myocardial tissue after cardiopulmonary resuscitation in rats / 中华危重病急救医学

Objective To investigate the changes in cyclophilin A (CyPA) signal pathway in rat myocardial tissue after cardiopulmonary resuscitation (CPR).Methods Sixty-eight healthy male Sprague-Dawley (SD) rats were randomly divided into sham operation group (n =8),instantancous CPR after cardiac arrest (CA),immediate CPR after CA (CRP),and 15,30,60 and 120 minutes after CPR groups,respectively,with 10 rats in each group.Asphyxia was simulated by occlusion of the tracheal tube at the end of exhalation.Mechanical ventilation,compression and epinephrine injection were given for restoration of spontaneous circulation (ROSC) in order to reproduce cardiopulmonary resuscitation after cardiac arrest (CA-CPR) models in rats.Hemodynamic changes were recorded at different time points.The blood was collected from abdominal aorta,and myocardial tissue was also harvested.The serum CyPA and CD147 levels were determined by enzyme-linked immunosorbent assay (ELISA).The protein positive cells and mRNA expression of CyPA and CD147 in myocardial tissue of the rats were determined by immunohistochemical and real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR).The protein expressions of matrix metalloproteinases (MMP-2,MMP-9) and myeloperoxidase (MPO) in rat myocardial tissue were determined by Western Blot.The neutrophil infiltration in rat myocardial tissue was assessed by hematoxylin and eosin (HE) staining.Results The heart rate of rat was lowered to 0 with arterial pressure lowered immediately after CA.Arterial pressure was elevated to normal range immediately after CPR.Heart rate was restored at about 30 minutes later,and the dose of epinephrine was 50-60 μg,and ROSC time was 1-4 minutes.Compared with those of the sham group,serum CyPA and CD147 levels were gradually increased along with elongation of ROCS time within 120 minutes,and peaked at 120 minutes,CyPA was increased from (786.11 ± 3.93) μg/L to (2 001.80 ± 10.61) μg/L,and CD147 was increased from (2.94±0.02) μg/L to (5.99±0.023) μg/L (P ＜ 0.05 or P ＜ 0.01).CyPA and CD147 mRNA expressions (A value) in rat myocardial tissue were gradually increased,and peaked at 120 minutes.Relative expression of CyPA at 120 minutes was 2.42 ± 0.05 when it was 1 in the control,and relative expression of CD147 at 120 minutes was 1.88 ±0.10 (both P ＜ 0.01).The immunohistochemical results under light microscope showed that the brown positive cells were gradually increased,which indicated that the expressions of CyPA and CD147 were increased.Expressions of MMP-2,MMP-9 and MPO (gray value) in myocardial tissue were also gradually increased,peaking at 120 minutes,and MMP-2 was increased from 0.396 ± 0.021 to 0.879 ± 0.020,MMP-9 was increased from 0.372 ± 0.009 to 0.819±0.012,and MPO was increased from 0.176±0.005 to 0.829±0.018 (P ＜ 0.05 or P ＜ 0.01).No obvious neutrophil infiltration in myocardial tissue was found with HE staining.Conclusion Expressions of CyPA and CD 147 were up-regulated in serum and myocardial tissue after CPR in rats,which may be the markers of inflammatory reaction.

Expression changes of cyclophilin A,Krüppel-like factor 2,and endothelial nitric oxide synthase in the pathological process of atherosclerosis in rats / 中国脑血管病杂志

Objective To observe the expression changes of cyclophilin A ( CyPA ) , Krüppel-like factor 2 (KLF2),and endothelial nitric oxide synthase (eNOS) in the pathological process of atherosclerosis in rats. Methods Thirty-two male Wistar rats were randomly divided into four groups:a control group and atherosclerosis 8,12,and 15 week groups (n=8 in each group). The atherosclerosis groups were fed with high-fat diet,and they were given vitamin D3 600 000 IU for a single intraperitoneal injection,and the rats in the control group were fed with the basal feed. The rats were sacrificed at 8,12,and 15 weeks. Their abdominal aortas were separated and stained with hematoxylin-eosin staining. Immunohistochemical method was used to detect the expression levels of CyPA,KLF2,and eNOS on arterial walls. Results With the progression of atherosclerosis lesion,the rats in the control group,and atherosclerosis 8,12,and 15 week groups showed different characteristics, including normal vessel structure, endothelial cell damage, smooth muscle cell proliferation,formation of atherosclerotic plaque,and formation of calcified plaque. There were significantly differences in the expression levels of CyPA,KLF2,eNOS on vessel walls among the control group,atherosclerosis 8,12,and 15 week groups (CyPA:0. 0037 ± 0. 0018,0. 0276 ± 0. 0090,0. 0526 ± 0. 0129,and 0. 1080 ± 0. 0388;KLF2:0. 0932 ± 0. 0331,0. 0415 ± 0. 0076,0. 0207 ± 0. 0059, and 0. 0014 ± 0. 0003;eNOS:0. 0358 ± 0. 0185,0. 0148 ± 0. 0080,0. 0049 ± 0. 0025,0. 0037 ± 0. 0026;F=36. 395,42. 108,and 21. 255,respectively,all P<0. 05),and the CyPA expression showed an increasing trend among groups, and there were significant differences between the pairwise comparisons (all P<0. 05). The expression level of KLF2 showed a progressive declining trend,and there were significant differences between pairwise comparisons (all P <0. 05);there was significant difference in eNOS expression between the atherosclerosis 12-week group and the atherosclerosis 15-week group, and there were significant differences among other groups (all P<0. 05). Conclusion With the progression of atherosclerotic lesions, the expression of CyPA increases progressively, and the expression levels of KLF2 and eNOS decrease progressively. This may be one of the approaches of CyPA caused atherosclerotic lesion.

Correlation of serum cyclophilin A level with occurance of essential hypertension in Xinjiang Xibo and Mongolia populations and its clinical significance / 天津医药

Objective To investigate the correlation of cyclophilin A (CyPA) level in patients with essential hyperten?sion(EH)in Xinjiang Xibo and Mongolia populations and to investigate its clinical significance. Methods EH patients (n=76) and healthy control subjects (n=62) were selected from Xinjiang Xibo populations. On the other hand, EH patients (n=66)and healthy control subjects (n=57) were selected from Xinjiang Mongolia populations. Serum cyclophilin A was measured using enzyme-linked immunosorbent assay(ELISA). Parameters such as age, body mass index(BMI), waist, waist hip ratio, blood pressure were measured in each group. The correlation of cyclophilin A with each parameter was analyzed. Results The levels of serum cyclophilin A in EH patients in Xinjiang Xibo and Xinjiang Mongolia populations were significantly high?er than those in their corresponding healthy control groups(P<0.05). Serum cyclophilin A in patients with essential hyperten?sion(EH)in Xinjiang Mongolia population was signifantly higher than that in EH patients from Xinjiang Xibo population (P<0.05). There was a positive correlation between serum cyclophilin A level and occurance of EH in both Xinjiang Xibo and Xinjiang Mongolia populations. Conclusion The serum cyclophilin A is significant higher in EH patients in both Xinji?ang Xibo and Xinjiang Mongolia populations so it is positively correlated with occurance of EH (P<0.05). Cyclophilin A lev?el is a risk factor of essential hypertension in both Xinjiang Xibo and Xinjiang Mongolia populations.

Cloning and expression of cyclophilin A gene from Echinoccocus granulosus / 中华检验医学杂志

Objective In order to assess the value of Echinoccocus granulosus cyclophilin A (EgypA) in immune diagnosis,this novel gene was cloned and expressed.Methods By screening the EST library,the coding region of EgCypA was identified,and the PCR primers were designed based on this sequence.Bioinformatic tools were used to deduce the amino acid sequence of EgCypA and analyzed its biological characteristics.EgCypA was amplified from E.granulosus cDNA library by using PCR.Then,it was cloned into the prokaryotic expression vector pET28a and transformed into E.coli BL21.The recombinant protein was expressed in E.coli BL21 after IPTG induction.The immunogenicity of EgCypA was evaluated by Western blot using the Echinoccocus granulosus and other parasites infected animals' sera.Results Results of SDS-PAGE electrophoresis show that the recombinant BL21 expressed 18 400-25 000 protein,which was identical with the molecular weight calculated by bioinformatic analysis.Western blot shows that the recombinant protein only reacted with its immune serum and E.granulosus cystic fluid immune serum,and EgCypA immune serum could react with the excretion and secretion antigen from E.granulosus protoscolexes,and no cross-reaction between EgCypA and sera from other parasites infected animals.Conclusions Cloning and expression of EgCypA are successful.EgCypA has good immunologic activity and could be a candidate molecular for immune diagnosis of echinococcosis in early stage.

Serum CyPA level change in patients with coronary heart disease and its clinical significance / 重庆医学

Objective To observe the serum cyclophilin A(CyPA)level change in the patients with coronary heart disease (CHD)and to investigate its clinical significance.Methods 64 patients with CHD(CHD group)were divided into three groups ac-cording to the clinical types:stable angina pectoris(SAP)group,unstable angina pectoris(UAP)group and acute myocardial infarc-tion(AMI)group;which were divided into three groups according to the coronary artery lesion range:single vessel lesion group, double vessels lesions group and multi vessels lesions group;while 26 controls with normal coronary arteries were select as the con-trol group.Serum levels of CyPA and matrix metalloproteinase-9 (MMP-9 )were detected by ELISA,and C-reactive protein(CRP) concentration was detected by the immune scattering turbidimetry test.Results The CyPA,MMP-9 and CRP levels in the CHD group were significantly higher than those in the control group(P0.05);serum CyPA,MMP-9 and CRP levels in the single vessel lesion group,the double vessels lesion group,multi vessels lesion group were significantly higher than those in the control group(P<0.05),serum CyPA,MMP-9 and CRP levels were elevated with the increase of coronary artery lesion vessels (P<0.05).In the CHD group,serum CyPA with MMP-9 and CRP showed the significantly positive correlation(r=0.772,0.749, P<0.01).Conclusion Serum CyPA level is significantly increased in the patients with CHD,CyPA may have some relationship to CHD and the plaque stability.

Análise da expressão imuo-histoquímica da ciclofilina A, EMMPRIN e MMP-7 na doença periodontal / Expression analysis imuo staining of cyclophilin A, EMMPRIN and MMP-7 in periodontal disease

A doença periodontal é uma entidade infecciosa que resulta da resposta imuno-inflamatória do hospedeiro aos microrganismos presentes no biofilme dentário, levando à destruição tecidual. O propósito do presente estudo foi avaliar a expressão imuno-histoquímica da ciclofilina A (CYPA), do indutor de metaloproteinases da matriz extracelular (EMMPRIN) e da metaloproteinase da matriz 7 (MMP-7) em espécimes humanos de gengiva clinicamente saudável (n=32), gengivite induzida por biofilme dentário (n=28) e periodontite crônica (n=30). Foram realizadas biópsias das três condições clínicas e feita a análise imuno-histoquímica através da contagem total do número de células positivas, correlacionando-a com parâmetros clínicos. A imunopositividade da CYPA, do EMMPRIN e da MMP-7 revelou diferença estatisticamente significativa entre os três grupos, com maiores percentuais de positividade nos espécimes de periodontite crônica, seguidos pelos espécimes de gengivite crônica e de gengiva saudável (p < 0,001). Foi evidenciada maior expressão de CYPA e MMP-7 nos grupos que tinham infiltrado inflamatório mais intenso. Foram observadas correlações das imunoexpressões de EMMPRIN, MMP-7 e CYPA, tanto entre si como com parâmetros clínicos (profundidade de sondagem e perda de inserção clínica). Foram verificadas correlações positivas entre a expressão de CYPA e as expressões da MMP-7 (r = 0,831; p < 0,001) e do EMMPRIN (r = 0,289; p = 0,006). Além disso, a profundidade de sondagem revelou correlação positiva, estatisticamente significativa, com as expressões de MMP-7 (r = 0,726; p < 0,001), EMMPRIN (r = 0,345; p = 0,001) e CYPA (r = 0,803; p < 0,001). Esses resultados evidenciam que a CYPA, o EMMPRIN e a MMP-7 podem estar associadas à patogênese e progressão da doença periodontal. (AU)

Periodontal disease is an infectious disease resulting from the immunoinflammatory response of the host to microorganisms present in the dental biofilm which causes tissue destruction. The objective of this study was to evaluate the immunohistochemical expression of cyclophilin A (CYPA), extracellular matrix metalloproteinase inducer (EMMPRIN) and matrix metalloproteinase 7 (MMP-7) in human specimens of clinically healthy gingiva (n=32), biofilm-induced gingivitis (n=28), and chronic periodontitis (n=30). Immunopositivity for CYPA, EMMPRIN and MMP-7 differed significantly between the three groups, with higher percentages of staining in chronic periodontitis specimens, followed by chronic gingivitis and healthy gingiva specimens (p < 0.001). Immunoexpression of CYPA and MMP-7 was higher in the intense inflammatory infiltrate observed mainly in cases of periodontitis. Analysis of possible correlations between the immunoexpression of EMMPRIN, MMP-7 and CYPA and between the expression of these proteins and clinical parameters (probing depth and clinical attachment loss) showed a positive correlation of CYPA expression with MMP-7 (r = 0.831; p < 0.001) and EMMPRIN (r = 0.289; p = 0.006). In addition, there was a significant positive correlation between probing depth and expression of MMP-7 (r = 0.726; p < 0.001), EMMPRIN (r = 0.345; p = 0.001), and CYPA (r = 0.803; p < 0.001). These results suggest that CYPA, EMMPRIN and MMP-7 are associated with the pathogenesis and progression of periodontal disease. (AU)