17?-estradiolum On Attenuating Propagation Of Human Pulmonary Artery Smooth Muscle Cells Via ER?

Objective: Observe the role of 17?-Estradiolum on influencing the propagation of human pulmonary artery smooth muscle cells from cell level in hypoxic pulmonary hypertension?HPH?.Further investigate whether estrogen receptor??ER??or estrogen receptor??ER??can mediate it.Methods:1 Cell culture: The normoxia group of hPASMCs placed in incubator under 37 ? ? 5% CO₂ and 95%O₂ for cultivating.The hypoxia group of hPASMCs placed in incubator under 37 ? ? 5% CO₂ and 92%N2 for cultivating.Every 2 day change fluid for one time.With 0.25% trypsin digestion of subculture,cells in logarithmic phase,the experiments for 5⁸cells.Before the intervention of E2,the experimental cells are kept without FBS withoutphenol red DMEM culture training for 24 h.2 Cell intervention: The 5⁸ generations of hPASMCs were used,they were given normoxia group,hypoxia group and hypoxia+E2?10-6mol/L,10-7mol/L,10-8mol/L,10-9mol/L?group,detected the prolification of hPASMCs under different conditon by MTT for chosing the best dose of intervence.Then hPASMCs were given normoxia group,hypoxia group and hypoxia+E2?10-6mol/L?group,hypoxia+E2?10-6mol/L?+MPP group,hypoxia+E2?10-6mol/L?+PHTPP group.Detected the prolification of hPASMCs under different conditon by MTT.These groups were evaluated by RT-PCR method to explore the expression of ER alpha,beta and PCNA mRNA levels in hPASMCs,under the condition of different interventions were evaluated by Western blot method to explore the expression of ER alpha,beta and PCNA protein levels in hPASMCs.Results:Chronic hypoxia obviously promoted the proliferation of hPASMCs?P<0.01?.proliferation of hPASMCs decreased after intervened by E2,the best dose of E2 were 10-6mol/L.Moreover Chronic hypoxia obviously promoted the proliferation of hPASMCs?P<0.01?with the decreasing of expression of ER?mRNA and protein?P<0.01?,increasing of expression of PCNAmRNA and protein?P<0.01?,while the expression of ER?mRNA and protein did not change obviously?P>0.05?;In the goup of hypoxia+E2,the proliferation of hPASMCs was decreased?P<0.01?and the change of ER??PCNAmRNA and protein was reversed?P<0.01?,while ER?did not?P>0.05?.In the group of hypoxia+E2+MPP,the result which was in goup of hypoxia+E2 was weakened?P<0.01?.While in the group of hypoxia+E2+PHTPP,the result which was in goup of hypoxia+E2 did not change obviously?P>0.05?.Conclusion:1 E2 can effectively attenuate the poliferation of the hPASMCs2 E2 can attenuate the poliferation of the hPASMCs via ER?,further attenuating the development of HPH.