| Objective:The purpose of this study was to explore the relationship between PPAR ? signal pathway,CYP4 A and 20-HETE and renal damage in spontaneously hypertensive rats,and to observe the effects of acupuncture on key factors such as PPAR ?,CYP4 A,in renal tissue and serum 20-HETE,serum creatinine and blood urea nitrogen in spontaneously hypertensive rats,and to explore the possible mechanism of acupuncture intervention on renal damage in spontaneously hypertensive rats.Methods:30 SPF grade homologous 10-week-old healthy male spontaneously hypertensive rats weighing 180-220 g and 10 normotensive SPF-grade Wistar rats weighing 180-220 g were selected.Using SPSS24.0 software,30 spontaneously hypertensive rats were randomly divided into Taichong group,Quchi group and model group,and normal Wistar rats were divided into normal group.In Taichong group,acupuncture at Taichong acupoint,Zusanli acupoint and Hegu acupoint on the left side of SHR were inserted vertically at 90 °,lifted and twirled uniformly clockwise,followed by flat tonifying,relieving and reducing manipulation,indwelling for 15 minutes,and then twirled counterclockwise.Quchi group acupuncture SHR left Quchi point,Zusanli point and Hegu point,the operation is the same as before.The rats in the normal group and model group were fixed for 15 minutes without acupuncture or other operations.The blood pressure was measured 1 week before treatment,1 day before treatment and 28 days after treatment.The rats were placed in the fixator,and after they were quiet,they were operated in a quiet state,measured repeatedly for 3 times,and the average value was taken.At the end of acupuncture treatment on the 28 th day,fasting could not abstain from water for 24 hours.before sampling,blood samples were anesthetized with 5%1.5g/kg.Blood samples were taken from the abdominal aorta.ELISA method was used to detect the expression levels of serum 20-HETE,serum creatinine and serum urea nitrogen.Kidney tissue was taken to detect the gene expression of PPAR ?,CYP4A1 and CYP4A2/3 in different groups by RT-PCR method.Western-Blot method was used to determine the expression of PPAR ? and CYP4 A protein.Renal tissue sections were made,and HE staining was used to observe the basic morphological changes of kidneys in each group.Results:(1)Comparison of blood pressure in each group: one week before treatment,compared with one day before treatment,there was no significant change in blood pressure in normal group,model group,Taichong group and Quchi group(P > 0.05).Compared with the model group,the blood pressure in the a Taichong group and Quchi group decreased significantly(P<0.05),and the blood pressure in the Taichong group and Quchi group decreased significantly before and after treatment(P<0.05).(2)Comparison of the concentration of 20-HETE in serum of rats in each group: after 4weeks of treatment,there was no significant difference in 20-HETE between the Taichong group and the normal group(P>0.05),but the level of 20-HETE in the Taichong group was significantly lower than that in the model group(P<0.05).(3)Comparison of the content levels of serum creatinine and serum urea nitrogen in each group: after 4 weeks of treatment,the content levels of serum creatinine and serum urea nitrogen in Taichong group and Quchi group were significantly lower than those in the model group(P < 0.05).There was no significant difference between serum creatinine and serum urea nitrogen(P>0.05).(4)Comparison of the levels of PPAR ?-m RNA,CYP4A1-m RNA and CYP4A2/3-m RNA in each group: after 4 weeks of treatment,compared with the model group,PPAR ? in Taichong group and Quchi group was significantly up-regulated,while CYP4A1 and CYP4A2/3-m RNA in Taichong group and Quchi group weredown-regulated and significantly different(P < 0.05).Compared with Quchi group CYP4A1 and CYP4A2/3-m RNA in Taichong group were significantly lower than those in Quchi group(P<0.05).(5)Comparison of PPAR ? and CYP4 A protein expression in each group:(1)PPAR ?:after 4 weeks of treatment,compared with the model group,the expression of PPAR ?protein in Taichong group and Quchi group was significantly higher than that in Quchi group(P<0.05),but there was no significant difference in PPAR ? protein expression between Taichong group and Quchi group(P>0.05).(2)CYP4A: after 4 weeks of treatment,compared with the model group,the expression of CYP4 A protein in Taichong group and Quchi group was down-regulated and there was significant difference(P<0.05),while there was no significant difference in CYP4 A protein expression between Taichong group and Quchi group(P>0.05).(6)Comparison of HE staining results of renal tissue of rats in each group: after HE staining,there was no significant change in the distribution of glomerulus,renal tubule and renal medulla in the normal group,but there were significant changes in the renal tissue structure in the model group compared with the normal group: glomerulus atrophy,glomerular telangiectasia,hyperemia and uneven distribution of renal medulla.Compared with the model group,the renal tissue of Taichong group and Quchi group had a slight improvement in glomerular atrophy and hyperemia,and the distribution of renal medulla was more uniform and denser than that in the model group.Conclusion:(1)Acupuncture can effectively reduce the blood pressure of SHR.(2)The improvement effect of many experimental indexes in Taichong group was better than that in Quchi group,which indicated that the acupuncture effect of Taichong point,Hegu point and Zusanli point was better.(3)Acupuncture can effectively reduce the content of SHR Scr and BUN and improve the damage of renal target organs in SHR.(4)Acupuncture can activate the expression of PPAR ?,and the inhibition of CYP4A/20-HETE,may be one of the mechanisms affecting the damage of renal target organs. |
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