| 1164 Hyperglycemia Upregulates Selectin Expression in the Gingiva of Akita Mice | ||
|
K. RHOURIDA, T. VAN DYKE, and R. GYURKO, Boston University, MA, USA OBJECTIVE: Leukocyte rolling and attachment is increased in gingival vessels of the diabetic Akita mice, as we have previously shown using gingival intravital microscopy. Since selectins play a prominent role in the initial steps of leukocyte attachment to the vascular endothelium, we have monitored P-selectin expression in the gingival tissues of wild-type and Akita mice. METHODS: Under ketamine/xylazine anesthesia gingival inflammation was induced in wild-type and Akita mice by injecting TNFα (100 ng each quadrant) or PBS as control. Two hours later mice were euthanized and gingival tissue was collected. Gingival protein extracts (60 μg/lane) were loaded on 6% SDS-polyacrylamide gel and electrophoresed at 120 V for 1 hour. Proteins were transferred to a PVDF membrane and incubated with P-selectin antibody (1:200 dilution) overnight. Secondary antibody was applied for 2 hours (1:5000 dilution). Finally, protein bands were detected with luminal-enhanced chemiluminescence detection reagent, followed by autoradiography. P-selectin levels are expressed as relative band intensity as determined with densitometry. RESULTS: P-selectin was detected at its expected molecular weight as a ~140 kDa band. Under baseline conditions (PBS injection only) Akita P-selectin signal was 44 ± 8 % higher than the wild-type. Following TNFα injection both wild-type and Akita P-selectin signal was enhanced (wild-type: 111 ± 80 %, Akita: 139 ± 79 % increase compared to PBS-injected gingiva), and Akita gingival P-selectin expression was 64 ± 7 % higher when compared to that of wild type. CONCLUSION: These results indicate that chronic hyperglycemia enhances the expression of P-selectin in the absence or in the presence of inflammation. Increased P-selectin expression is likely to contribute to the enhanced leukocyte rolling and attachment observed in Akita gingival vessels. Moreover, enhanced selectin expression may contribute to the pro-inflammatory phenotype observed in the periodontium of diabetic patients. Supported by DE014568 (NIDCR), DE016933 (NIDCR) and DE015566 (NIDCR). | ||
| Seq #126 - Microbes and Oral Infections 3:30 PM-4:45 PM, Thursday, March 22, 2007 Ernest N. Morial Convention Center Exhibit Hall I2-J | ||
|
Back to the Microbiology / Immunology and Infection Control Program
| ||