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1.
Fig. 2

Fig. 2. Dose-response of LPS-induced increases in serum levels of sVCAM-1 and sICAM-1 (A), MCP-1 (B), and TNFα (C) in wild-type mice. From: Genetic Deficiency of NADPH Oxidase Does Not Diminish but Rather Enhances LPS-Induced Acute Inflammatory Responses in Vivo.

Wild-type mice were injected i.p. with HBSS (control) or different doses of LPS as described in Methods. Three hours after LPS injection, the animals were sacrificed and blood was collected. Serum sVCAM-1, sICAM-1, MCP-1, and TNFα were measured by ELISA. Data shown are mean values ± SEM of 4–8 animals.

Wei-Jian Zhang, et al. Free Radic Biol Med. ;46(6):791-798.
2.
Fig. 7

Fig. 7. LPS-induced endotoxemic death in wild-type, gp91phox−/− (A), and p47phox−/− mice. From: Genetic Deficiency of NADPH Oxidase Does Not Diminish but Rather Enhances LPS-Induced Acute Inflammatory Responses in Vivo.

Wide-type (closed circles), gp91phox−/− (open circles, panel A) and p47phox−/− mice (open circles, panel B) were injected i.p. with 1.5 mg LPS as described in Methods. The animals were monitored for survival three times daily for up to five days. The numbers of animals used were: 10 and 19 wild-type mice, respectively, in Panels A and B; 10 gp91phox−/− mice; and 11 p47phox−/− mice.

Wei-Jian Zhang, et al. Free Radic Biol Med. ;46(6):791-798.
3.
Fig. 5

Fig. 5. Time-course of LPS-induced activation of lung NFκB and AP-1 in wild-type mice. From: Genetic Deficiency of NADPH Oxidase Does Not Diminish but Rather Enhances LPS-Induced Acute Inflammatory Responses in Vivo.

Wild-type mice were injected i.p. with HBSS (0 time point control) or 50µ g LPS as described in Methods. One, 3, and 24 hours after LPS injection, the animals were sacrificed and nuclear extracts were isolated from lung. NFκB (p65)/DNA (closed circles) and AP-1 (c-fos)/DNA (open circles) binding activities were quantified by ELISA. Data were expressed as fold of HBSS-treated control and are presented as mean values ± SEM of 4–8 animals.

Wei-Jian Zhang, et al. Free Radic Biol Med. ;46(6):791-798.
4.
Fig. 1

Fig. 1. Time-course of LPS-induced increases in serum levels of sVCAM-1 (A), sICAM-1 (B), MCP-1 (C), and TNF (D) in wild-type and gp91phox−/− mice. From: Genetic Deficiency of NADPH Oxidase Does Not Diminish but Rather Enhances LPS-Induced Acute Inflammatory Responses in Vivo.

Wild-type (solid circles) and gp91phox−/− (open circles) mice were injected i.p. with HBSS (0 time point control) or 50µ g LPS as described in Methods. At the indicated time points after LPS injection, the animals were sacrificed and blood was collected. Serum sVCAM-1, sICAM-1, MCP-1, and TNFα were measured by ELISA. Data shown are mean values ± SEM of 4–8 animals.

Wei-Jian Zhang, et al. Free Radic Biol Med. ;46(6):791-798.
5.
Fig. 4

Fig. 4. Dose-response of LPS-induced increases in lung VCAM-1 and ICAM-1 (A), MCP-1 and IL-6 (B), IL-1β (C), and TNFα (D) mRNA levels in wild-type mice. From: Genetic Deficiency of NADPH Oxidase Does Not Diminish but Rather Enhances LPS-Induced Acute Inflammatory Responses in Vivo.

Wild-type mice were injected i.p. with HBSS (control) or different doses of LPS as described in Methods. Three hours after LPS injection, the animals were sacrificed and total RNA was isolated from lung. Real-time quantitative PCR analysis was performed for VCAM-1, ICAM-1, MCP-1, TNFα, IL-1β, IL-6, and GAPDH mRNA. Data were expressed as fold of HBSS-treated control after normalization to the internal control gene, GAPDH, and are presented as mean values ± SEM of 4–8 animals.

Wei-Jian Zhang, et al. Free Radic Biol Med. ;46(6):791-798.
6.
Fig. 6

Fig. 6. LPS-induced increases in myeloperoxidase mRNA levels in various organs of wild-type, gp91phox−/− (A), and p47phox−/− (B) mice. From: Genetic Deficiency of NADPH Oxidase Does Not Diminish but Rather Enhances LPS-Induced Acute Inflammatory Responses in Vivo.

Wild-type, gp91phox−/−, and p47phox−/− mice were injected i.p. with HBSS (control) or 50µ g LPS as described in Methods. Three hours after LPS injection, the animals were sacrificed and total RNA was isolated from various organs. Real-time quantitative PCR analysis was performed for MPO and GAPDH mRNA. After normalization to the internal control gene, GAPDH, the results for the MPO gene were expressed as fold of control. Data shown are mean values ± SEM of 4–8 animals. *P<0.05 compared to WT mice

Wei-Jian Zhang, et al. Free Radic Biol Med. ;46(6):791-798.
7.
Scheme 1

Scheme 1. Diagram depicting the TLR4 and NOX-ROS pathways. From: Genetic Deficiency of NADPH Oxidase Does Not Diminish but Rather Enhances LPS-Induced Acute Inflammatory Responses in Vivo.

LPS binds to LPS-binding protein (LBP) and CD14, thus activating TLR4. TLR4 activates two major kinases: IκB kinase (IKK), which phosphorylates IκB, leading to its ubiquitylation and subsequent degradation and NFκB translocation to the nucleus; and mitogen-activated protein kinase kinases (MKK), which phosphorylate and activate c-Jun kinase (JNK), extracellular receptor-activated kinase (ERK), and p38 kinase, leading to AP-1 activation. Activated NFκB and AP-1 in the nucleus bind to DNA promoter regions to induce inflammatory gene transcription. In the NOX-ROS pathway, LPS activates TLR4, which in turn activates NADPH oxidase through Rac1 and Nox4. While it has been hypothesized that ROS generation by NADPH oxidase enhances IKK and MKK activity (arrows marked “?”), the results presented in this paper indicate that the NOX-ROS pathway contributes to the resolution of LPS-induced inflammation.

Wei-Jian Zhang, et al. Free Radic Biol Med. ;46(6):791-798.
8.
Fig. 3

Fig. 3. Time-course of LPS-induced increases in lung VCAM-1 (A), ICAM-1 (B), MCP-1 (C), TNFα (D), IL-1β (E), and IL-6 (F) mRNA levels in wild-type and gp91phox−/− mice. From: Genetic Deficiency of NADPH Oxidase Does Not Diminish but Rather Enhances LPS-Induced Acute Inflammatory Responses in Vivo.

Wild-type (solid circles) and gp91phox−/− (open circles) mice were injected i.p. with HBSS (0 time point control) or 50µ g LPS as described in Methods. At the indicated time points after LPS injection, the animals were sacrificed and total RNA was isolated from lung. Real-time quantitative PCR analysis was performed for VCAM-1, ICAM-1, MCP-1, TNFα, IL-1β, IL-6, and GAPDH mRNA. Data were expressed as fold of HBSS-treated control after normalization to the internal control gene, GAPDH, and are presented as mean values ± SEM of 4–8 animals. *P<0.05 compared to WT mice.

Wei-Jian Zhang, et al. Free Radic Biol Med. ;46(6):791-798.

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