Hydrogen Sulfide On Cell Proliferation Of CFs In Rats Induced By AVP And Its Mechanism Study

Objective：To study the effect of hydrogen sulfide on cell proliferation of therat cardiac fibroblasts (CFs) in rats induced by vasopressin(AVP)and its relationship with calcineurin.Methods：1. Cell culture and processing: recovery and batches.2.Cells in cultrue: The rat cardiac fibroblasts were cultured anddivided into12holes that each hole contains cell number5x103,including AVP group、NaSH groups、AVP+NaSH groups and control group.NaSH groups include A, B, C, D, E five hole which Concentration of10-9mol/L,10-8mol/L,10-7mol/L,10-6mol/L,10-5mol/L respectively.AVP+NaSH groups also include A, B, C, D, E five hole which all have10-7mmol/L AVP processing and its concentration of NaSH same the NaSHintervention group; AVP group is processed by10-7mmol/L AVP. afterculture and the intervention of72hours,Each cell leave thesupernatant and collection of cells.3. Determined by MTT testdetect the absorbance value of each rat cardiac fibroblastsfor understanding of cell proliferation4. Taking AVP+NaSH groupsabsorbance value of the minimum concentration of hole、NaSH group、AVP group and control group, by flow cytometry technique evaluateDNA synthesis of rat cardiac fibroblasts percentage period(S).5.Taking AVP+NaSH groups absorbance value of the minimum concentrationof hole、NaSH group、AVP group and control group, by spectrophotometerevaluate Calcineurin activity. Result：1.AVP group’s CFs absorbance value(1.382±0.012) significantlyovertoped the control group (1.058±0.015)(P<0.05)； AVP+NaSH setof each concentration gradient in the cultivation of hole in theabsorbance of CFs: Respectively（1.313±0.010）、（1.248±0.014）、（1.229±0.014）、（1.197±0.009）、（1.136±0.009）,the E hole ofabsorbance value than other each hole in AVP+NaSH groups arereduced(all P<0.05)；but AVP group’s CFs absorbance value(1.382±0.007)significantly overtopped the AVP+NaSH groups (均P<0.05)；Respectively（1.070±0.013）、（1.053±0.012）、（0.94±0.011）、（0.912±0.011）、（0.75±0.012）,the A, B, C, D, E holes of NaHS groupsdeclined in comparison with control group(1.172±0.009)(P<0.05);2.(1)S-phase cell fraction in AVP group（35.86%）and AVP+NaSH group（E）（24.07%） significantly overtop the control group(22.51%)(allP<0.05）；but the AVP+NaSH group(E) low the AVP group（24.07%VS35.86%，P<0.05）；the NaSH group(E) S-phase cell fraction low thecontrol group（18.47%VS22.51%，P<0.05）.3.(1)Calcineurin activity in AVP goup(74.214±5.72)pg/mg and AVP+NaSH group（E）（59.011±3.65）pg/mg was significantly higher thanthe control group(45.773±4.756)pg/mg(all P<0.01)；but Calcineurinactivity in the NaSH+AVP group（E）was significantly lower than theAVP goup (P<0.05)； calcineurin activity in the NaSH group（E）（8.578±2.074）pg/mg significantly reduced to the control group(P<0.05).Conclusion：1、Vasopressin can induce the proliferation of rat cardiacfibroblasts；2、Hydrogen sulfide can inhibit cardiac fibroblasts proliferation byAVP;3、The influence of hydrogen sulfide to vasopressin mediating rat cardiac fibroblasts proliferation may be related to its inhibitionof calmodulin phosphatase level.