Objective:

2

To investigate cerebral vasorelaxation of total flavonoids of

Chuzhou chrysanthemum

(TFCC) in rats and its mechanism.

Methods:

2

Cerebral basilar arteries (CBA) of rats were isolated

and the vasodilation induced by TFCC (10–2

560 mg/L) following pretension with 100 nmol/L U

46619

or 30 mmol/L KCl were measured using a pressure myograph system. Addition of H

2

S synthase cystathionine-γ-lyase (CSE) inhibitor dl-propargylglycine (PPG

100 μmol/L)

a large-conductance Ca

2+

-activated potassium (BK

Ca

) channel blocker iberiotoxin (IBTX

100 nmol/L) and L-type Ca

2+

channel blocker nifedipine (100 nmol/L) were added to determine the effect of pretreatment with the inhibitors on TFCC-induced vasorelaxation. KCl (30 mmol/L)

was used as a contractile agent

TFCC (3.3–270 mg/L)-induced relaxation was detected by measuring the length of the long axis of rat cerebral vascular smooth muscle cells (VSMCs). Determination of the effect of pretreatment of VSMCs by IBTX or nifedipine on TFCC-induced cellular relaxation

and intracellular free Ca

2+

concentration([Ca

2+

]

i

) was detected by fluorescent method. Endothelial cells (ECs) were co-cultured with VSMCs to observe the effect of endogenous H

2

S on TFCC-induced relaxation in VSMCs.

Results:

2

TFCC caused a concentration-dependent vasorelaxation in the rat CBA precontracted with KCl or U

46619

(

P

<

0.01). Endothelial removal markedly attenuated this vasodilation

but the remaining vasorelaxation was still significant (

P

<

0.01). The TFCC-induced cerebral vasorelaxation was remarkably inhibited by PPG

IBTX and nifedipine (

P

<

0.01). TFCC caused a significant relaxation of rat CBA VSMCs (

P

<

0.01). Co-culture with wild-type cerebral ECs but not with cystathionine-γ-lyase-or 3-mercaptosulfotransferase-knockout ECs markedly enhanced TFCC-induced relaxation of VSMCs (

P

<

0.05) and increased H

2

S content (

P

<

0.01). TFCC decreased the [Ca

2+

]

i

in VSMCs (

P

<

0.01)

which was attenuated by PPG and IBTX.

Conclusions:

2

TFCC dilated rat CBA in both endothelium-dependent and -independent manner. Its endothelium-dependent dilation was probably involved in the blockade of L-type Ca

2+

channels caused by endothelial H

2

S activating BK

Ca

channels in VSMCs; its endothelium-independent relaxation was primarily from the direct blockade of the L-type Ca

2+

channels in VSMCs.