ACTHR in regulating aldosterone secretion to angiotensin Ⅱ and potassium chloride stimulation / 中华内分泌外科杂志

ABSTRACT

Objective To study CYP11B2 mRNA and aldosterone secretion alteration in human adrenocortical carcinoma H295R cell after angiotensin Ⅱ (AT-Ⅱ) and potassium chloride stimulation,and to investigate the effect of adrenocorticotropic hormone receptor (ACTHR) on them.Methods Lentiviral vector was used to increase ACTHR expression.It was transfected into the H295R cells.Similarly,another H295R cells,without ACTHR vector,was used as the control group.ACTHR alteration was measured by Western blot and real-time polymerase chain reaction (RT-PCR).CYP11B2 mRNA was detected at 24 hours after 100 nmol/L AT-Ⅱ/16 mmol/L KCL stimulation,and the amplification of the two groups was compared.Aldosterone was measured by ELISA kit.Results Compared with those in control ceils,the protein and mRNA level of ACTHR in experimental cells were increased 2.4 times and 18 times respectively (P＜0.05).CYP11B2 mRNA of experimental cells was 1.7 times higher than control cells after 24 h stimulation of AT-Ⅱ.Aldosterone production was 121.98+8.31 and 104.05+6.88 ng/L respectively.The former amplification was 2.06 times higher than that of the latter (P＜0.05).Similarly,CYP11B2 mRNA of experimental cells was 19.2 times higher than control cells after 24 h stimulation of KCL.Aldosterone production was 137.67±10.35 and 104.05 ± 6.88 ng/L respectively.The former amplification was 3.13 times higher than that of the latter (P＜0.05).Conclusion Overexpression of ACTHR increases the sensitivity and response of CYP11B2 mRNA and aldosterone to AT-Ⅱ and KCL stimulation,and ACTHR is expected to become a key protein in understanding primary aldosteronism.