Staphylococcus aureus biofilm influences the expression of lysozyme, SLPI and gp340 in a human sinonasal explant model / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the influences of staphylococcus aureus in planktonic and biofilm forms on the expression of lysozyme, SLPI and gp340 in the human sinonasal explant model.@*METHOD@#Mucosa samples from ethmoid sinus were collected from ten patients of cerebrospinal fluid leak and were cultured with and without S. aureus biofilms and planktonic cells. After the infection, the explant model was confirmed by CLSM, and the secretion of lysozyme, SLPI and gp340 was detected by enzyme-linked immunosorbent assay (ELISA) at 8, 16, and 24 h after S. aureus challenge. Expressions of lysozyme, SLPI and gp340 in mRNA and protein levels after 24 h S. aureus challenge were detected using RT-PCR, immunohistochemistry and Western bolt assay respectively.@*RESULT@#The secretion of lysozyme, SLPI and gp340 in the explant model was observed with a trend to increase in a time-dependent manner. At 8 and 16 h after S. aureus challenge, the secretion of lysozyme, SLPI and gp340 in biofilms group was significantly higher than these in planktonic cells group and control group (P 0.05), the biofilms enhanced the expressions of lysozyme, SLPI and gp340 significantly compared with planktonic cells and controls (P < 0.05).@*CONCLUSION@#S. aureus biofilm induced the expressions of lysozyme, SLPI and gp340 to a higher level than planktonic cells, indicating that S. aureus biofilm was an influencing factor on the innate immune system.

Nonoxynol-9 berberine plural gel has little effect on expression of SLPI, SP-D and lactoferrin in mice's vagina

The most frequently used spermicide Nonoxynol-9 [N-9] in the clinic alters the vaginal flora, which will result in an increased risk of opportunistic infection. So development of a novel spermicidal and microbicidal drug appears to be inevitable. Vaginal local immune is an important part of vaginal flora. Secretory leukocyte protease inhibitor [SLPI], surfactant proteins D [SP-D], and lactoferrin [LF] are anti-microbial molecules with important roles in immune system of female vaginas. To observe effect of a vaginal spermicide nonoxynol-9 [N-9] berberine plural gel on the expression of SLPI SP-D and LF in mice's vaginas. Female BABL/C mice were randomly divided into following 5 groups: normal control group, blank gel group, berberine gel group, 12% N-9 gel group and N-9 berberine plural gel group. Estradiol benzoate at physiological dose was done by hypodermic injection to every group's mice. After 72h, drug gels were separately injected into the mice's vaginas, while immunohistochemistry and Western blot were taken to detect the expression of the 3 indexes in mice's vaginas respectively after 24h and 72h of gel injection. The differences in the three indexes between normal control group and blank gel group were not significant statistically [p>0.05]. The expression of the three indexes in 12% N-9 gel group was decreased compared to that in blank gel group [p<0.05]. The differences in the three indexes between N-9 berberine plural gel group and blank gel group were not significant statistically [p>0.05]. Also, the three index's level of 24h and 72h in sub observation groups after treatment were without statistical significance [p>0.05]. Application of N-9 berberine plural gel had little impact on antimicrobial peptides in normal mice's vaginas

Expression and clinical significance of secretory leucocyte protease inhibitor in colon carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of secretory leucocyte protease inhibitor (SLPI) in colon cancer and their clinical significance.</p><p><b>METHODS</b>Immunohistochemistry was performed to detect the SLPI expression in colon cancer tissue microarray. The expression of SLPI was scored by two pathologists and was analyzed using Χ(2) test to explore its influence on the pathologic characteristics of colon carcinoma.</p><p><b>RESULTS</b>SLPI was up-regulated in colon cancer tissue compared to normal mucosa. Overexpression of SLPI protein was correlated with differentiation grade (low differentiation: 42.1% vs 57.9%; moderate/well differentiation: 2.3% vs 97.7%, TNM stages(III-IV:29.4% vs 70.6%;I-II:3.1% vs 96.9%), lymph node metastasis (28.6% vs 71.4%) and distant metastasis (84.6% vs 15.4%), but not with patient age or sex.</p><p><b>CONCLUSION</b>SLPI overexpression correlates with aggressive pathologic characteristics of colon cancer and it may server as prognostic factor of colon cancer patients. Further research will be carried out to verify whether SLPI can become a new target for colon cancer treatment.</p>

Application of genome-wide microarray for screening genes related to peritoneal metastasis of colorectal cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen genes related to peritoneal metastasis of colorectal cancer.</p><p><b>METHODS</b>Specimens of primary cancer and normal mucosa tissues were collected from 3 patients with peritoneal metastasis of colorectal cancer. The total RNA were extracted and inversely transcribed into cDNA to synthesize aRNA using in vitro RNA synthesis. The synthesized aRNA, after labeling with Cy3, were hybridized with the whole human genome oligo microarray. The Empirical Bayes method was used to screen the differentially expressed genes, followed by confirmation of the selected genes by semi-quantitative RT-PCR.</p><p><b>RESULTS</b>With a threshold of P≤0.05, a total of 105 differentially expressed genes were identified in primary cancer lesions, including 42 up-regulated and 63 down-regulated genes. Three of the up-regulated genes (S100P, PRDX1 and SLPI) were selected and confirmed by RT-PCR, which yielded results consistent with those from gene microarray.</p><p><b>CONCLUSION</b>Gene microarray technique can provide valuable clues for locating the tumor markers of peritoneal metastasis in colorectal cancer patients.</p>

The change of secretory leukocyte protease inhibitor and its correlation with elastase in the development of periodontal inflammation / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the change of secretory leukocyte protease inhibitor (SLPI) and elastase (EA) in the different stages of periodontal inflammation and to evaluate the possibility of the two proteins as saliva markers reflecting overall periodontal health status.</p><p><b>METHODS</b>Unstimulated whole saliva were collected from 86 subjects (divided into 4 groups as healthy, gingivitis, moderate periodontitis and severe periodontitis). Fifteen patients with moderate or severe periodontitis were only given scaling and root planning (SRP). Whole saliva was collected and clinical patameters were recorded at baseline and four weeks after the treatment. SLPI concentrations were determined with enzyme linked immunosorbent assay (ELISA) systems, while EA with low-molecular-weight substrate reaction.</p><p><b>RESULTS</b>There were no statistical differences of SLPI concentrations among four groups (P > 0.05). However, EA activities in moderate periodontitis and severe periodontitis groups [0.077 (0.060) and 0.077 (0.489)] were higher than in healthy and gingivitis group [0.058 (0.028) and 0.058 (0.024)] (P < 0.05). SLPI only showed a weak negative correlation with age (r = -0.301, P < 0.05), rather than with EA or clinical parameters. In 15 patients with chronic periodontitis the mean concentration of SLPI and EA activity was 2.031 (2.449) µg/L and 0.075 (0.118), and both decreased significantly to 1.405 (0.659) µg/L and 0.055 (0.028) respectively 4 weeks after SRP.</p><p><b>CONCLUSIONS</b>After SRP, the decrease of SLPI concentration and EA activity in saliva may reflect the periodontal inflammation subsiding. SLPI in saliva was not correlated with the development of periodontal inflammation.</p>

Proteomic and Transcriptomic Analysis of Interleukin-1beta Treated Airway Epithelial Cells / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Mucin hypersecretion is one of the main symptoms of inflammatory diseases in the respiratory tract. The authors previously reported that pleiotypic pro-inflammatory cytokine, interleukin (IL)-1beta, plays significant roles in the respiratory tract inflammation by inducing mucins (MUC2, MUC5AC, MUC8). However, the molecular mechanism for mucin hypersecretion in the respiratory tract is still unclear. MATERIALS AND METHOD: In order to understand the mechanisms of mucin hypersecretion in the airway epithelium, the differentially expressed proteins and genes in the lung mucoepidermoid carcinoma cell line (NCI-H292 cells), which were treated for 6 and 24 hours with IL-1beta (10 ng/ml), were identified using 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE) proteomics and cDNA microarray analysis (8.6 K). RESULTS: In the 2-D PAGE, 8 differentially expressed proteins and 14 post-translational modification proteins were identified 6 and 24 hrs after the IL-1beta treatment. Microarray analysis identified a total of 413 genes (6.6%) in the 6-hour treatment group and 115 genes (2.0%) in the 24-hour treatment group that were regulated after the IL-1beta treatment. The differentially expressed genes that were regulated by the IL-1beta treatment were mostly found in the metabolic pathway rather than in the regulatory pathway. A comparison of the proteomic and microarray data showed that there was a large discrepancy between the protein expression and the gene expression levels. Among the genes encoding the proteins secreted in the airway, MUC5B was down-regulated but sialomucin CD 164, lysozyme, and the secretory leukocyte protease inhibitor (SLPI) were up-regulated. CONCLUSION: These results clearly show that the transcript levels have little value in predicting the extent of protein expression. Genomics and proteomics have different evaluation fields. Therefore, they may not provide all the information on the gene and protein profiles.

Morphological Differences of the Wound Healing in Secretory Leukocyte Protease Inhibitor Knockout Mice / 체질인류학회지

Secretory leukocyte protease inhibitor (SLPI) is a serine protease inhibitor with anti-microbial properties found in mucosal fluids. At the tissue level, the ability of this 12kDa protein is to counteract the excessive degradation of functional and structural proteins such as collagen and fibronectin. Impaired healing states are characterized by excessive proteolysis and often bacterial infection, leading to the hypothesis that SLPI may have a role in this process. To investigate the role of SLPI in skin how it contributes to tissue repair, we have generated mice null for the gene encoding SLPI (Slpi), which show impaired cutaneous wound healing with increased inflammation. For the purpose of this, we have performed wound experiment in skin tissue with morphometrical analyses, immunohistochemistry, and Rnase protection assay. From these analyses, the results were that delayed healing in KO mice wounds compared to that of WT, prolonged inflammatory phase and increased TGF-beta1 in KO wounds, and lower mechanical properties in KO wounds. Taken together, SLPI may play a cruical role in cutaneous wounds healing especially in matrix reorganization that suggests the development as a clinical drug for wound healing.

Regulation of Mucin and Non-Mucin Secretions and Gene Expression by Triiodothyronine and Collagen Gel in Human Airway Epithelium / 대한이비인후과학회지

BACKGROUND AND OBJECTS: We have been interested in elucidating the role of hormones and growth factors in regulating differentiation and mucin and non-mucin secretions. Our purpose is to investigate the effects of each supplement contained in the culture medium for mucin and non-mucin secretions. MATERIALS AND METHODS: Individual factors were removed from the culture media of normal human tracheobronchial epithelial (NHTBE) cells grown in air-liquid interface cultures. The effects on the cell phenotype, mucin, lysozyme (LZ), and secretory leukocyte protease inhibitor (SLPI) secretion and gene expression were examined. RESULTS: Deletion of hydrocortisone, epinephrine, transferrin or amphotericin-gentamycin from the media had no reproducible effects; Deletion of insulin was incompatible with culture growth. Removal of triiodothyronine selectively increased mucin secretion, but did not affect the gene expression. However, MUC5AC mRNA levels were reproducibly increased, suggesting that the expressions of these two mucin genes were differentially regulated. LZ and SLPI secretion levels were not significantly affected by the deletion of triiodothyronine from the culture media. The LZ mRNA levels were increased in the absence of triiodothyronine whereas the SLPI transcript levels were not affected. Omission of the attachment substratum and the type 1 collagen gel resulted in a significant increase in all 3 secretory products. MUC2 and MUC5AC steady state mRNA levels were not consistently affected. In contrast, LZ and SLPI gene expressions were reproducibly increased. CONCLUSION: This study shows that individual factors in the epithelial environment can regulate the expression of specific secretory cell gene products in a highly selective manner.

Regulation of Mucin and Non-Mucin Secretions and Gene Expression by Retinoic Acid in Human Airway Epithelium / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Airway hypersecretion is a frequent feature of several respiratory tract diseases including rhinitis, sinusitis, and otitis media. Efforts are being made in several laboratories to elucidate mechanisms involved in the regulation of secretion. There are several factors which modulate expression of the secretory phenotype, such as retinoic acid (RA), triiodothyronine, steroid, and extracellular matrix. We have been interested in elucidating the role of retinoids in regulating differentiation of mucin and non-mucin secretions. MATERIALS AND METHODS: Retinoic acid was removed from the culture media of normal human tracheobronchial epithelial cells grown in the air-liquid interface cultures. The effects on cell phenotype and mucin, lysozyme (LZ), and the secretory leukocyte protease inhibitor (SLPI) secretion and gene expression were examined. RESULTS: Removal of RA from the media induced squamous differentiation and caused a drastic decrease in mucin secretion and a decrease in expression of the mucin genes, MUC2 and MUC5AC. Lysozyme and SLPI secretions were increased in RA-depleted cultures. Paradoxically, LZ mRNA was decreased, while the SLPI mRNA levels were increased. A most intriguing finding was the paradoxical response of LZ to RA-depletion. The reason for this apparant incongruity between mRNA and protein levels is currently under investigation. CONCLUSION: Our studies show that RA is an important factor for mucous differentiation.