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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

Baicalin-induced apoptosis in immature and mature DCs in a time-dependent way. Immature and mature DCs were treated with 50 μM of BA for different time (24–72 h), then cells were harvested and apoptosis was assayed by flow cytometry, using annexin V/PI staining. Representative results of three independent experiments were presented.
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Figure 3: Baicalin-induced apoptosis in immature and mature DCs in a time-dependent way. Immature and mature DCs were treated with 50 μM of BA for different time (24–72 h), then cells were harvested and apoptosis was assayed by flow cytometry, using annexin V/PI staining. Representative results of three independent experiments were presented.

Mentions: We further compared BA-induced apoptosis in mature DCs and immature DCs. In this experiment, the day-6 DCs stimulated with LPS were used as mature DCs, because they highly expressed CD80/CD86 and secreted high level of IL-12. In comparison, the day-6 DCs without stimulation by LPS were used as immature DCs. Mature and immature DCs were treated with 50 μM of BA for different time (24–72 h) or with graded concentrations of BA (12.5–50 μM) for 48 h, then cellular apoptosis was assessed by FACS after annexin V/PI staining. BA-induced apoptosis in both mature and immature DCs in time- and dose-dependent way and immature DCs were more sensitive for BA-induced apoptosis than mature DC (Figures 3 and 4).


Baicalin induced dendritic cell apoptosis in vitro.

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

Baicalin-induced apoptosis in immature and mature DCs in a time-dependent way. Immature and mature DCs were treated with 50 μM of BA for different time (24–72 h), then cells were harvested and apoptosis was assayed by flow cytometry, using annexin V/PI staining. Representative results of three independent experiments were presented.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: Baicalin-induced apoptosis in immature and mature DCs in a time-dependent way. Immature and mature DCs were treated with 50 μM of BA for different time (24–72 h), then cells were harvested and apoptosis was assayed by flow cytometry, using annexin V/PI staining. Representative results of three independent experiments were presented.
Mentions: We further compared BA-induced apoptosis in mature DCs and immature DCs. In this experiment, the day-6 DCs stimulated with LPS were used as mature DCs, because they highly expressed CD80/CD86 and secreted high level of IL-12. In comparison, the day-6 DCs without stimulation by LPS were used as immature DCs. Mature and immature DCs were treated with 50 μM of BA for different time (24–72 h) or with graded concentrations of BA (12.5–50 μM) for 48 h, then cellular apoptosis was assessed by FACS after annexin V/PI staining. BA-induced apoptosis in both mature and immature DCs in time- and dose-dependent way and immature DCs were more sensitive for BA-induced apoptosis than mature DC (Figures 3 and 4).

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