Limits...
Qingkailing Suppresses the Activation of BV2 Microglial Cells by Inhibiting Hypoxia/Reoxygenation-Induced Inflammatory Responses.

Mana L, Wang S, Zhu H, Xing Y, Lou L, Wu A, Dong B, Sun Y, Yang S, Wang L, Gao Y - Evid Based Complement Alternat Med (2014)

Bottom Line: Our previous experiments confirmed that QKL exerts an inhibitory effect on cerebral ischemia-induced inflammatory responses.In this study, BV2 microglial cells were used to validate the protective effects of QKL treatment following ischemia-reperfusion injury simulated via hypoxia/reoxygenation in vitro.However, QKL treatment also displayed dose-dependent differences in its inhibitory effects on p38 phosphorylation and inflammatory factor expression.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine, Beijing 100700, China.

ABSTRACT
Qingkailing (QKL) is a well-known composite extract used in traditional Chinese medicine. This extract has been extensively administered to treat the acute phase of cerebrovascular disease. Our previous experiments confirmed that QKL exerts an inhibitory effect on cerebral ischemia-induced inflammatory responses. However, whether QKL suppresses the activation of microglia, the primary resident immune cells in the brain, has yet to be determined. In this study, BV2 microglial cells were used to validate the protective effects of QKL treatment following ischemia-reperfusion injury simulated via hypoxia/reoxygenation in vitro. Under these conditions, high expression levels of ROS, COX-2, iNOS, and p-p38 protein were detected. Following ischemia/reperfusion injury, QKL significantly increased the activity of BV2 cells to approximately the basal level by modulating microglial activation via inhibition of inflammatory factors, including TNF- α , COX-2, iNOS, and p-p38. However, QKL treatment also displayed dose-dependent differences in its inhibitory effects on p38 phosphorylation and inflammatory factor expression.

No MeSH data available.


Related in: MedlinePlus

Effects of different durations of hypoxia/reoxygenation on BV2 microglia cells. (a) BV2 cells were exposed to hypoxia for 6 or 12 hours followed by reoxygenation for 12 hours. Morphological changes were observed under an inverted microscope (×100). (b) After exposure to hypoxia for 12 hours, the BV2 cells were reoxygenated for 12 or 24 hours. Cell viability was assed using the MTT method. Each value indicates the mean ± SD. N = 5.  ##P < 0.01 compared to the control group. (c) ROS level in BV2 microglial cells after hypoxia for 12 hours and reoxygenation for 12 hours based on flow cytometry. N = 4.  ##P < 0.01 compared to the control group.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4020462&req=5

fig1: Effects of different durations of hypoxia/reoxygenation on BV2 microglia cells. (a) BV2 cells were exposed to hypoxia for 6 or 12 hours followed by reoxygenation for 12 hours. Morphological changes were observed under an inverted microscope (×100). (b) After exposure to hypoxia for 12 hours, the BV2 cells were reoxygenated for 12 or 24 hours. Cell viability was assed using the MTT method. Each value indicates the mean ± SD. N = 5.  ##P < 0.01 compared to the control group. (c) ROS level in BV2 microglial cells after hypoxia for 12 hours and reoxygenation for 12 hours based on flow cytometry. N = 4.  ##P < 0.01 compared to the control group.

Mentions: Based on inverted phase contrast microscopy, approximately 60–80% of the cells in the control group were well-anchored contained, a polygonal cell body; approximately 20–40% of the cells were suspended. As hypoxia continued, the cell body gradually became rounded, and the attached cells became suspended and gathered into clusters. Hypoxia for 12 h resulted in significant changes compared to the control group, and these changes were exacerbated after reoxygenation for 24 h (Figure 1(a)). MTT colorimetry suggested that, compared to the control group, after hypoxia for 12 h followed by reoxygenation for 12 h or 24 h in an three-gas incubator, the number of surviving BV2 cells in the model group were significantly reduced (P < 0.01) (Figure 1(b)). As shown in Figure 1(c), the level of ROS in the model group (hypoxia for 12 h and reoxygenation for 12 h) was significantly higher than the normal group (P < 0.01).


Qingkailing Suppresses the Activation of BV2 Microglial Cells by Inhibiting Hypoxia/Reoxygenation-Induced Inflammatory Responses.

Mana L, Wang S, Zhu H, Xing Y, Lou L, Wu A, Dong B, Sun Y, Yang S, Wang L, Gao Y - Evid Based Complement Alternat Med (2014)

Effects of different durations of hypoxia/reoxygenation on BV2 microglia cells. (a) BV2 cells were exposed to hypoxia for 6 or 12 hours followed by reoxygenation for 12 hours. Morphological changes were observed under an inverted microscope (×100). (b) After exposure to hypoxia for 12 hours, the BV2 cells were reoxygenated for 12 or 24 hours. Cell viability was assed using the MTT method. Each value indicates the mean ± SD. N = 5.  ##P < 0.01 compared to the control group. (c) ROS level in BV2 microglial cells after hypoxia for 12 hours and reoxygenation for 12 hours based on flow cytometry. N = 4.  ##P < 0.01 compared to the control group.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4020462&req=5

fig1: Effects of different durations of hypoxia/reoxygenation on BV2 microglia cells. (a) BV2 cells were exposed to hypoxia for 6 or 12 hours followed by reoxygenation for 12 hours. Morphological changes were observed under an inverted microscope (×100). (b) After exposure to hypoxia for 12 hours, the BV2 cells were reoxygenated for 12 or 24 hours. Cell viability was assed using the MTT method. Each value indicates the mean ± SD. N = 5.  ##P < 0.01 compared to the control group. (c) ROS level in BV2 microglial cells after hypoxia for 12 hours and reoxygenation for 12 hours based on flow cytometry. N = 4.  ##P < 0.01 compared to the control group.
Mentions: Based on inverted phase contrast microscopy, approximately 60–80% of the cells in the control group were well-anchored contained, a polygonal cell body; approximately 20–40% of the cells were suspended. As hypoxia continued, the cell body gradually became rounded, and the attached cells became suspended and gathered into clusters. Hypoxia for 12 h resulted in significant changes compared to the control group, and these changes were exacerbated after reoxygenation for 24 h (Figure 1(a)). MTT colorimetry suggested that, compared to the control group, after hypoxia for 12 h followed by reoxygenation for 12 h or 24 h in an three-gas incubator, the number of surviving BV2 cells in the model group were significantly reduced (P < 0.01) (Figure 1(b)). As shown in Figure 1(c), the level of ROS in the model group (hypoxia for 12 h and reoxygenation for 12 h) was significantly higher than the normal group (P < 0.01).

Bottom Line: Our previous experiments confirmed that QKL exerts an inhibitory effect on cerebral ischemia-induced inflammatory responses.In this study, BV2 microglial cells were used to validate the protective effects of QKL treatment following ischemia-reperfusion injury simulated via hypoxia/reoxygenation in vitro.However, QKL treatment also displayed dose-dependent differences in its inhibitory effects on p38 phosphorylation and inflammatory factor expression.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine, Beijing 100700, China.

ABSTRACT
Qingkailing (QKL) is a well-known composite extract used in traditional Chinese medicine. This extract has been extensively administered to treat the acute phase of cerebrovascular disease. Our previous experiments confirmed that QKL exerts an inhibitory effect on cerebral ischemia-induced inflammatory responses. However, whether QKL suppresses the activation of microglia, the primary resident immune cells in the brain, has yet to be determined. In this study, BV2 microglial cells were used to validate the protective effects of QKL treatment following ischemia-reperfusion injury simulated via hypoxia/reoxygenation in vitro. Under these conditions, high expression levels of ROS, COX-2, iNOS, and p-p38 protein were detected. Following ischemia/reperfusion injury, QKL significantly increased the activity of BV2 cells to approximately the basal level by modulating microglial activation via inhibition of inflammatory factors, including TNF- α , COX-2, iNOS, and p-p38. However, QKL treatment also displayed dose-dependent differences in its inhibitory effects on p38 phosphorylation and inflammatory factor expression.

No MeSH data available.


Related in: MedlinePlus