Zhu Jing,Xu Zihui.Mechanism of adenosine triphophate sensitive potassium channel in INS-1 cells mediated by diazoxide-interfered oxidative stress[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(11):1155-1158.
Zhu Jing,Xu Zihui.Mechanism of adenosine triphophate sensitive potassium channel in INS-1 cells mediated by diazoxide-interfered oxidative stress[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(11):1155-1158.DOI:
Mechanism of adenosine triphophate sensitive potassium channel in INS-1 cells mediated by diazoxide-interfered oxidative stress
Objective To study the effect of diazoxide (DIZ)-interfered oxidative stress on expression of SUR1 subunit gene in adenosine triphosphate sensitive potassium (KATP) channel of INS-I cells. Methods INS-1 cell strains were divided into oxidative stress group
DIZ interfere group
taurine interfere group
and control group. H
2
O
2
was added into INS-1 cell strains of oxidative stress group at the concentration of 50 μmol/L
100 μmol/L
200 μmol/L
and 400 μmol/L respectively
and cultured for 3 h. DIZ was added into INS-1 cell strains of DIZ interfere group at the concentration of 50 μmol/L and incubated for 30 min
then DIZ was added at the concentration of 400 μmol/L and incubated for an additional 3 h. Taurine was added into INS-1 cell strains of taurine interfere group at the concentration of 1μmol/L and incubated for 30 min
then H
2
O
2
was added at the concentration of 400 μmol/L and cultured for an additional 3 h. No DIZ
taurine
and H
2
O
2
were added into INS-1 cell strains of control group. Survival rate of INS-1 cell strains
glucose simulated insulin secretion (GSIS) level
and SUR1 subunit protein expression level were measured by MTT assay
ELISA
and Western blotting
respectively
in oxidative stress group
DIZ interfere group
and taurine interfere group. Results The survival rate of INS-1 cell strains [(105.27±3.83)%
]
was significantly higher in oxidative stress group than in control group after exposed to 50 μmol/L H
2
O
2
(P0.05) and gradually decreased with the increased concentration of H
2
O
2
. The survival rate of INS-1 cell strains was 68.66% and 51.67%
respectively
in oxidative stress group after exposed to 200 μmol/L a
nd 400 μmol/L of H
2
O
2
which was significantly lower than that in control group (P0.05). The GSIS level was higher in oxidative stress group than in control group after exposed to 50 μmol/L H
2
O
2
(118.47±10.15 mU/L vs 112.87±9.28 mU/L
P0.05) and gradually decreased with the increased concentration of H
2
O
2
. The GSIS level was significantly lower in oxidative stress group than in control group after exposed to 200 μmol/L and 400 μmol/L of H
2
O
2
(64.59±4.53 mU/L vs 85.79±7.03 mU/L
P0.05). The expression level of SUR1subunit protein in INS-1 cells was higher in oxidative stress group than in control group after exposed to 50 μmol/L H
2
O
2
for 3 h and significantly higher in DIZ group than in control group after exposed to 200 μmol/L and 400 μmol/L H
2
O
2
for 3 h(P0.05). However
no significant difference was observed between taurine interfere group and control group(P0.05). Conclusion DIZ inhibits the expression of SUR1 subunit gene in K
ATP
channel of INS-I cells by suppressing the release of ROS through inhibiting the electronic transport in respiration chain of mitochrondria
thus reducing the oxidative stress of INS-I cells.
Preconditioning with mitochondrial ATP-sensitive potassium channel opener, diazoxide, protects the myocardial mitochondial functions after ischemic injury in rats
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