Resistin Is Involved In Mediating H9c2 Cardiomyocyte Hypertrophy By Inhibiting Mir-489 Expression

Objective: To explore resistin-induced hypertrophy of H9c2 cardiomyocytes by inhibiting the expression of mi R-489..Methods: The H9c2 cardiomyocytes were cultured,and the experiments were carried out when the cells were in a good growth state.The experiment was divided into three parts according to the purpose of the experiment: The first part of the experiment was divided into four groups: Con group(blank control group): normal cultured cells,and the starvation medium was replaced with the experimental group at the same time;R50 group(resistin 50ng/ml group)),R100 group(resistin 100ng/ml group),R200 group(resistin 200ng/ml group): After the cells were seeded in six-well plates for 24 hours,the starvation medium was replaced for 24 hours,and then replaced with resistin containing 50ng/ml and 100ng/ml respectively.ml,200ng/ml starvation medium for 36 h.The second and third experiments were divided into four groups: Con group(blank control group):normal cultured cells,and the starvation medium was replaced at the same time as the experimental group;Res group(resistin group): cells were inoculated into six wells After 24 hours of plate,the starvation medium was changed for 24 hours,and then replaced with the starvation medium containing resistin 50ng/ml for 36 hours;mimic + Res group(mi R-489 mimic + resistin treatment group): cells were seeded in six-well plates for 24 hours and then treated with mi R-489 mimic transfected cells for 24 hours,then changed to starvation medium containing resistin 50ng/ml for 36 hours;NC + Res group(mimic-NC + resistin treatment group): cells were seeded in six-well plates for 24 hours and then transfected with mimic-NC The cells were stained for 24 h,and then replaced with a starvation medium containing 50 ng/ml of resistin for 36 h.Before the second part of the experiment,a preliminary experiment should be carried out to confirm that the transfection was successful.The pre-experiment was divided into the following two groups: mimic group: cells were seeded in six-well plates for 24 hours and then transfected with mi R-489 mimic for 24 hours;NC group: cells were seeded in six-well plates for 24 hours and cells were transfected with mimic-NC for 24 hours.The cell area was analyzed by Image J,the protein content of single cells was calculated after detection by microplate reader,and the relative expression levels of ANF,BNF and mi R-489 were detected by real-time fluorescence quantitative PCR.Results:(1)Within a certain concentration range,resistin could induce an increase in the surface area of H9c2 cardiomyocytes(248.860 ± 43.548 vs 93.131 ± 33.717,P < 0.001),and an increase in single cell protein content(2.516 ± 0.097 vs 1.499 ± 0.075,P < 0.01),The expressions of ANF and BNF were up-regulated(P < 0.001).When the concentration exceeds a certain concentration,although the surface area of cardiomyocytes still increases,the single-cell protein content and the expressions of ANF and BNF show a downward trend(single-cell protein content: R200 : R100 =2.726 ± 0.169 : 3.501 ± 0.281,P < 0.01 There was no significant difference in the relative expression of ANF and BNF in R200 group compared with blank group,P>0.05).(2)Compared with the Con group,the expression of mi R-489 in the three groups treated with resistin was significantly decreased(P < 0.05),and the expression of mi R-489 in the mimic+Res group was higher than that in the Res group.(3)Resistin can induce the increase of the surface area of H9c2 cardiomyocytes(236.714 ±27.861 vs.77.153 ± 11.341,P < 0.001),and the increase of single-cell protein content(3.455± 0.171 vs.2.221 ± 0.056,P < 0.01).The expression was up-regulated(P < 0.05),and overexpression of mi R-489 could significantly improve the above changes(P < 0.05).Conclusion:(1)Resistin can induce H9c2 cardiomyocyte hypertrophy;(2)Resistin can inhibit the expression of mi R-489;(3)Overexpression of mi R-489 ameliorated resistin-induced cardiomyocyte hypertrophy.In conclusion,the results obtained in this study show that resistin participates in the regulation of cardiomyocyte hypertrophy by inhibiting the expression of mi R-489,and overexpression of mi R-489 can improve it.