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  Indian J Med Microbiol
 

Figure 2: Lactoferrin inhibits cyclooxygenase.2 expression and prostaglandin E2 production. (a) Effect of lactoferrin (L) and nuclear factor.kappa B inhibitor (Bay 11–7082) on the gene expression of cyclooxygenase.2 in interleukin-1β .stimulated human osteoarthritis chondrocytes determined by quantitative real time-polymerase chain reaction. Primary human chondrocytes (70.80% confluent) were pretreated with L (50–75 μg/ml) or Bay 11–7082 (10 μM) for 2 h and stimulated with interleukin.1β (10 ng/ml) for 24 h. Expression of cyclooxygenase-2 mRNA was normalized to glyceraldehyde 3-phosphate dehydrogenase and compared to the levels present in control. Results are representative (mean ± standard error of mean) of duplicate experiments with chondrocytes obtained from osteoarthritis donors and values differ without a common letter P < 0.01. (b) Effect of lactoferrin and nuclear factor-kappa B inhibitor on the protein expression of cyclooxygenase-2 in interleukin-1β.stimulated human osteoarthritis chondrocytes. Primary chondrocytes were pretreated with L (50.75 μg/ml) for 2 h and stimulated with interleukin-1β (10 ng/ml) for 24 h and cell lysates were prepared. Cyclooxygenase-2 protein was analyzed by Western immunoblotting. Band images were digitally captured and the band intensities were obtained using the Un-Scan-It software and are expressed in average pixels of five independent scans. (c) Effect of lactoferrin and nuclear factor-kappa B inhibitor on the production of prostaglandin E2 in interleukin-1β.stimulated osteoarthritis chondrocytes culture medium. Primary chondrocytes were pretreated with L (50.75 μg/ml) for 2 h and stimulated with interleukin-1β (10 ng/ml) for 24 h. Prostaglandin E2 production was analyzed in cell culture supernatant by ELISA. Data shown are cumulative of three experiments and differ without a common letter P < 0.01

Figure 2: Lactoferrin inhibits cyclooxygenase.2 expression and prostaglandin E2 production. (a) Effect of lactoferrin (L) and nuclear factor.kappa B inhibitor (Bay 11–7082) on the gene expression of cyclooxygenase.2 in interleukin-1β .stimulated human osteoarthritis chondrocytes determined by quantitative real time-polymerase chain reaction. Primary human chondrocytes (70.80% confluent) were pretreated with L (50–75 μg/ml) or Bay 11–7082 (10 μM) for 2 h and stimulated with interleukin.1β (10 ng/ml) for 24 h. Expression of cyclooxygenase-2 mRNA was normalized to glyceraldehyde 3-phosphate dehydrogenase and compared to the levels present in control. Results are representative (mean ± standard error of mean) of duplicate experiments with chondrocytes obtained from osteoarthritis donors and values differ without a common letter <i>P</i> < 0.01. (b) Effect of lactoferrin and nuclear factor-kappa B inhibitor on the protein expression of cyclooxygenase-2 in interleukin-1β.stimulated human osteoarthritis chondrocytes. Primary chondrocytes were pretreated with L (50.75 μg/ml) for 2 h and stimulated with interleukin-1β (10 ng/ml) for 24 h and cell lysates were prepared. Cyclooxygenase-2 protein was analyzed by Western immunoblotting. Band images were digitally captured and the band intensities were obtained using the Un-Scan-It software and are expressed in average pixels of five independent scans. (c) Effect of lactoferrin and nuclear factor-kappa B inhibitor on the production of prostaglandin E2 in interleukin-1β.stimulated osteoarthritis chondrocytes culture medium. Primary chondrocytes were pretreated with L (50.75 μg/ml) for 2 h and stimulated with interleukin-1β (10 ng/ml) for 24 h. Prostaglandin E2 production was analyzed in cell culture supernatant by ELISA. Data shown are cumulative of three experiments and differ without a common letter <i>P</i> < 0.01