Effect Of Ellagic Acid On Hypoxic Pulmonary Hypertension

Objective: To study the effect of ellagic acid on hypoxic pulmonary hypertension and its possible mechanism.Methods: 1.Animal experiments and groupingForty SD male rats(180-220 g body weight)were randomly divided into four groups of 10 rats each.Respectively: A normoxic group: reared in normoxia at room temperature for 4weeks.B Normoxa + ellagic acid group: The ellagic acid 15 mg/kg was given to the rats daily under normoxic environment for 4 weeks.C.Hypoxia group: The rats were placed in a hypoxic chamber at 10% oxygen concentration for 8 hours at room temperature for four consecutive weeks.D Hypoxia + ellagic acid group: Before exposure to hypoxia in rats at room temperature,ellagic acid 15 mg/kg was intragastrically administered and then in a hypoxic chamber at 10% oxygen concentration for 8 hours for 4 consecutive weeks.2.Determination of right ventricular systolicpressure After the rats were treated as above for 8 weeks,a catheter was inserted into the right ventricle through the right jugular vein Theotherendisconnectedwithapressuresensor,andtherightventricularsystolic pressure(RVSP)is measured by a physiological signal recorder analysis system.3.Determination of right ventricular hypertrophyindex Take the experimental rats out of the heart,remove the left and right atrium and separate the right ventricle(RV)and left ventricular + ventricular septum(S +LV)and accurately weighed.The right ventricular hypertrophy index was calculated using the formula(RVHI)=(RV)/(LV+S).4.Determination of superoxide dismutase(SOD)and malondialdehyde(MDA)in lung tissue: Weigh 100 mg of lung tissue from the same site(lower right lung lobe)of each group of rats,grind to make tissue extract according to reagents.Box Description Step Determination of SOD and MDA content in lungtissue.5.Immunohistochemical staining of ?-SMA in lungtissue Lung tissues from the same part of the rats in each experimental group(lower left lobe)were used to make physiological sections of lung tissue and then stained with ?-SMAantibody.6.Rat pulmonary artery smooth muscle cells in vitro culture and experimental grouping Pulmonary arterial blood vessels from healthy rats were used for cell subculture and 3-5 generations of well-developed pulmonary arterial smooth muscle cells were harvested and grouped according to the following methods: A: normoxic +equal volume PBS solution group;B: hypoxic + equal volume PBS solution group C: hypoxia +5?mol/L ellagic acid group;D: hypoxia +10?mol/L ellagic acid group;E: hypoxia +15?mol/L ellagic acid group;F: hypoxia +20?mol/L floret flower Acid group.The grouped cells were incubated in a constant temperature incubator at 37°C for 48 hours.7.Trypan blue assay Trypsin-digested each group of cultured cells to make acell suspension.The cell concentration was adjusted to 1×105/ml,and the cell suspension was dropped at the ratio of 9:1 to the cell suspension/% 0.4 trypan blue solution.Hope blue solution,mix.The viable and dead cells in each group were counted under a microscope.8.Western-blot assay detected the expression of p38 MAPK and ERK1/2protein in each group ofcells.Results: 1.Compared with normoxic and normoxic + ellagic acid group rats,right ventricular hypertrophy index [RV/(LV+S)] was significantly higher in hypoxia group(p<0.05).Right heart hypertrophy index in hypoxic group was higher than that in hypoxic + ellagic acid group(p<0.05).There was no significant difference in right heart hypertrophy index between normoxic group and normoxic + ellagic acid group.(p>0.05).2.Compared with rats in normoxic group and normoxic + ellagic acid group,right ventricular hypertrophy index [RV/(LV+S)] increased significantly(p<0.05)in hypoxic rats,and hypoxia alone The right heart hypertrophy index of the rats was higher than that of the hypoxic + ellagic acid group(p<0.05).There was no significant difference in right heart hypertrophy index between normoxic and normoxic + ellagic acid groups(p)>0.05).3.Observing changes in pulmonary blood vessels after ?-SMA staining revealed that the pulmonary vascular structures significantly changed in normoxic rats compared with normoxic + ellagic acid rats.Compared with the normoxic group,pulmonary vascular intima of the hypoxic group was significantly thickened.Compared with rats in hypoxic + ellagic acid group,the pulmonary vascular media thickness decreasedsignificantly.4.Compared with the normoxic group,the SOD content in the hypoxic group was significantly lower(p<0.05).Compared with the hypoxic + ellagic acid group,the hypoxic group was compared with the hypoxic group.The SOD content was lower in the tissue(p<0.05).There was no significant difference in the SOD content between the normoxic group and normoxic + ellagic acid group(p>0.05).5.Compared with normoxic group rats,MDA content in hypoxic group rats increased significantly(p<0.05),compared with hypoxia + ellagic acid group rats,hypoxia + ellagic acid group The content of MDA in lung tissue of rats was significantly decreased(p<0.05).There was no significant difference in the content of MDA in lung tissue between normoxic and normoxic and ellagic acid groups(p>0.05).6.Compared with normoxic pulmonary arterial smooth muscle cells,ROS in pulmonary arterial smooth muscle cells in hypoxic group was significantly increased(p<0.05).ROS levels in pulmonary arterial smooth muscle cells in hypoxia + ellagic acid group were significantly higher than those in pulmonary arterial smooth muscle cells in hypoxic group alone.Decline(p<0.05).7.Compared with normoxic group,the expression of p38 MAPK and ERK1/2 protein in pulmonary arterial smooth muscle cells in hypoxic rats was significantly higher(p<0.05),compared with hypoxic group,hypoxic + ellagic acid The expression of p38 MAPK and ERK1/2 protein in the pulmonary artery smooth muscle cells of rats was significantly decreased(p<0.05).