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Baicalin induced dendritic cell apoptosis in vitro.

Zhang H, Jiao Q, Gong Q, Zhang Y, Zhang W, Hu Z - Front Pharmacol (2011)

Bottom Line: This study was aimed to investigate the effects of baicalin (BA), a major flavonoid constituent found in the herb Baikal skullcap, on dendritic cells (DCs).Exposure of DCs to BA (2-50 μM) during BM cell differentiation showed no effects on the up-regulation of CD80/CD86 expression on DCs in response to LPS stimulation, but reduced DCs recovery by inducing apoptosis, and significantly inhibited the release of IL-12 to culture supernatants.These results demonstrate that BA induces selective apoptosis in immature DCs possibly through mitochondria-mediated pathway.

View Article: PubMed Central - PubMed

Affiliation: School of Pharmacy, Second Military Medical University Shanghai, China.

ABSTRACT
This study was aimed to investigate the effects of baicalin (BA), a major flavonoid constituent found in the herb Baikal skullcap, on dendritic cells (DCs). DCs were generated by culturing murine bone marrow (BM) cells for 6 days with granulocyte-macrophage colony-stimulating factor and interleukin (IL)-4, and lipopolysaccharide (LPS) was added on day 5 to stimulate DCs maturation. The expression levels of DC maturity markers (CD80/CD86) were assessed by flow cytometry using direct immunofluorescence method. IL-12 levels in the culture supernatants were assayed by ELISA. Apoptosis of DCs was analyzed by flow cytometry after annexin V/propidium iodide staining. The mitochondrial membrane potential (Δψ(m)) changes were measured by using the J-aggregate forming lipophilic cation 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide (JC-1). Exposure of DCs to BA (2-50 μM) during BM cell differentiation showed no effects on the up-regulation of CD80/CD86 expression on DCs in response to LPS stimulation, but reduced DCs recovery by inducing apoptosis, and significantly inhibited the release of IL-12 to culture supernatants. BA-induced DC apoptosis in a time- and dose-dependent way, and immature DCs were more sensitive for BA-induced apoptosis than mature DC. BA also induced Δψ(m) changes in DCs. These results demonstrate that BA induces selective apoptosis in immature DCs possibly through mitochondria-mediated pathway.

No MeSH data available.


Related in: MedlinePlus

The impact of BA on mitochondrial transmembrane potential in immature and mature DC. Immature and mature DCs were treated with BA (0–50 μM) for 6 h, stained with JC-1 dye probe, and analyzed by flow cytometry. Numbers in lower right quadrant indicate the percentage of cells that emit only green fluorescence which is attributed to depolarized mitochondrial membrane. Representative of three individual experiments with similar results are shown.
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Figure 5: The impact of BA on mitochondrial transmembrane potential in immature and mature DC. Immature and mature DCs were treated with BA (0–50 μM) for 6 h, stained with JC-1 dye probe, and analyzed by flow cytometry. Numbers in lower right quadrant indicate the percentage of cells that emit only green fluorescence which is attributed to depolarized mitochondrial membrane. Representative of three individual experiments with similar results are shown.

Mentions: One of the early critical events in apoptosis is the loss/disruption of mitochondrial membrane potential (Δψm) in the cells, which eventually causes the initiation and activation of apoptotic cascades. We sought to determine whether BA-treatment had any effect on the Δψm in DCs using JC-1 staining. Figure 5 shows treatment of DCs with BA resulted in dose-dependent increase in the proportion of green fluorescence-positive cells (indicating loss of mitochondrial membrane potential) in both immature DCs and mature DCs, and higher proportion of green fluorescence-positive cells were detected in BA-treated immature DCs. These results confirm that BA induced the loss of Δψm in DCs, which is more profound in immature DCs than in mature DCs.


Baicalin induced dendritic cell apoptosis in vitro.

Zhang H, Jiao Q, Gong Q, Zhang Y, Zhang W, Hu Z - Front Pharmacol (2011)

The impact of BA on mitochondrial transmembrane potential in immature and mature DC. Immature and mature DCs were treated with BA (0–50 μM) for 6 h, stained with JC-1 dye probe, and analyzed by flow cytometry. Numbers in lower right quadrant indicate the percentage of cells that emit only green fluorescence which is attributed to depolarized mitochondrial membrane. Representative of three individual experiments with similar results are shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3108619&req=5

Figure 5: The impact of BA on mitochondrial transmembrane potential in immature and mature DC. Immature and mature DCs were treated with BA (0–50 μM) for 6 h, stained with JC-1 dye probe, and analyzed by flow cytometry. Numbers in lower right quadrant indicate the percentage of cells that emit only green fluorescence which is attributed to depolarized mitochondrial membrane. Representative of three individual experiments with similar results are shown.
Mentions: One of the early critical events in apoptosis is the loss/disruption of mitochondrial membrane potential (Δψm) in the cells, which eventually causes the initiation and activation of apoptotic cascades. We sought to determine whether BA-treatment had any effect on the Δψm in DCs using JC-1 staining. Figure 5 shows treatment of DCs with BA resulted in dose-dependent increase in the proportion of green fluorescence-positive cells (indicating loss of mitochondrial membrane potential) in both immature DCs and mature DCs, and higher proportion of green fluorescence-positive cells were detected in BA-treated immature DCs. These results confirm that BA induced the loss of Δψm in DCs, which is more profound in immature DCs than in mature DCs.

Bottom Line: This study was aimed to investigate the effects of baicalin (BA), a major flavonoid constituent found in the herb Baikal skullcap, on dendritic cells (DCs).Exposure of DCs to BA (2-50 μM) during BM cell differentiation showed no effects on the up-regulation of CD80/CD86 expression on DCs in response to LPS stimulation, but reduced DCs recovery by inducing apoptosis, and significantly inhibited the release of IL-12 to culture supernatants.These results demonstrate that BA induces selective apoptosis in immature DCs possibly through mitochondria-mediated pathway.

View Article: PubMed Central - PubMed

Affiliation: School of Pharmacy, Second Military Medical University Shanghai, China.

ABSTRACT
This study was aimed to investigate the effects of baicalin (BA), a major flavonoid constituent found in the herb Baikal skullcap, on dendritic cells (DCs). DCs were generated by culturing murine bone marrow (BM) cells for 6 days with granulocyte-macrophage colony-stimulating factor and interleukin (IL)-4, and lipopolysaccharide (LPS) was added on day 5 to stimulate DCs maturation. The expression levels of DC maturity markers (CD80/CD86) were assessed by flow cytometry using direct immunofluorescence method. IL-12 levels in the culture supernatants were assayed by ELISA. Apoptosis of DCs was analyzed by flow cytometry after annexin V/propidium iodide staining. The mitochondrial membrane potential (Δψ(m)) changes were measured by using the J-aggregate forming lipophilic cation 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide (JC-1). Exposure of DCs to BA (2-50 μM) during BM cell differentiation showed no effects on the up-regulation of CD80/CD86 expression on DCs in response to LPS stimulation, but reduced DCs recovery by inducing apoptosis, and significantly inhibited the release of IL-12 to culture supernatants. BA-induced DC apoptosis in a time- and dose-dependent way, and immature DCs were more sensitive for BA-induced apoptosis than mature DC. BA also induced Δψ(m) changes in DCs. These results demonstrate that BA induces selective apoptosis in immature DCs possibly through mitochondria-mediated pathway.

No MeSH data available.


Related in: MedlinePlus