OBJECTIVE To investigate the impact of perilla alcohol on pulmonary vascular remod-eling in hypoxia-induced pulmonary hypertension(HPH)rats and to assess its regulatory effects on the angiotensin-converting enzyme 2(ACE2)-angiotensin(1-7)[Ang(1-7)]-mas proto-oncogene receptor(Mas)and ACE-angiotensinⅡ(AngⅡ)-angiotensin type 1 receptor(AT1R)axes.METHODS An HPH rat model was established by keeping them in a hypobaric chamber that simulated an altitude of 5 000 m for 28 d.Male SD rats were randomly divided into the control group,hypoxia group,hypoxia+sildenafil group(ig administration of 30 mg·kg-1),and hypoxia+perillyl alcohol groups(ig administration of 25,50 and 100 mg·kg-1 respectively).After 28 d,the levels of glutamic-oxaloacetic transaminase(GOT),glutamic-pyruvic transaminase(GPT),and blood urea nitrogen(BUN)in rat serum were measured to evaluate the toxic effects of perillyl alcohol on rat organs.Mean pulmonary artery pressure(mPAP)was measured via right ventricular catheterization.HE staining was used to observe the patho morphological changes of pulmonary vessels in HPH rats and measure the percentages of the vascularwall area[WA(%)],wall thickness[WT(%)],lumen area[LA(%)].The right ventricular hypertrophy level and α-smooth muscle actin(α-SMA)expression level were detected by immunohistochemistry and immunofluorescence.Western blotting was used to detect the protein expression levels of α-SMA,ACE,AT1R,AngⅡ,ACE2 and Mas in lung tissues of rats.Enzyme-linked immunosorbent assay(ELISA)was employed to mea-sure the content of Ang(1-7)in lung tissues.RESULTS Compared with the control group,the serum levels of GOT,GPT and BUN in the hypoxia group rats were significantly increased,but were signifi-cantly decreased by perillyl alcohol and sildenafil intervention.Compared with the control group,mPAP,WA(%),WT(%),right ventricular inner diameter(RVID),right ventricular free wall thickness(RVFWT)and fibrosis levels were significantly increased in the hypoxia group,while LA(%)was signifi-cantly decreased.Besides,the protein expression levels of α-SMA,ACE,AT1R and AngⅡ in lung tissues were significantly elevated while those of ACE2 and Mas,as well as Ang(1-7)content were signifi-cantly decreased in hypoxia group compared to the control group.Following intervention with perillyl alcohol and sildenafil,the protein expression levels of ACE and AngⅡin lung tissues of HPH rats were significantly decreased compared to the model group while those of ACE2 and Mas receptor,along with the content of Ang(1-7),were significantly increased.Perillyl alcohol significantly reduced the protein expression level of AT1R while sildenafil had no significant effect.CONCLUSION Perillyl alcohol can lower mPAP levels by improving pulmonary vascular remodeling and reducing pulmonary vascular fibrosis in HPH rats.The mechanism may be related to the regulatory effects on the ACE-AngⅡ-AT1R and ACE2-Ang(1-7)-Mas axes.

OBJECTIVE To investigate the impact of perilla alcohol on pulmonary vascular remod-eling in hypoxia-induced pulmonary hypertension(HPH)rats and to assess its regulatory effects on the angiotensin-converting enzyme 2(ACE2)-angiotensin(1-7)[Ang(1-7)]-mas proto-oncogene receptor(Mas)and ACE-angiotensinⅡ(AngⅡ)-angiotensin type 1 receptor(AT1R)axes.METHODS An HPH rat model was established by keeping them in a hypobaric chamber that simulated an altitude of 5 000 m for 28 d.Male SD rats were randomly divided into the control group,hypoxia group,hypoxia+sildenafil group(ig administration of 30 mg·kg-1),and hypoxia+perillyl alcohol groups(ig administration of 25,50 and 100 mg·kg-1 respectively).After 28 d,the levels of glutamic-oxaloacetic transaminase(GOT),glutamic-pyruvic transaminase(GPT),and blood urea nitrogen(BUN)in rat serum were measured to evaluate the toxic effects of perillyl alcohol on rat organs.Mean pulmonary artery pressure(mPAP)was measured via right ventricular catheterization.HE staining was used to observe the patho morphological changes of pulmonary vessels in HPH rats and measure the percentages of the vascularwall area[WA(%)],wall thickness[WT(%)],lumen area[LA(%)].The right ventricular hypertrophy level and α-smooth muscle actin(α-SMA)expression level were detected by immunohistochemistry and immunofluorescence.Western blotting was used to detect the protein expression levels of α-SMA,ACE,AT1R,AngⅡ,ACE2 and Mas in lung tissues of rats.Enzyme-linked immunosorbent assay(ELISA)was employed to mea-sure the content of Ang(1-7)in lung tissues.RESULTS Compared with the control group,the serum levels of GOT,GPT and BUN in the hypoxia group rats were significantly increased,but were signifi-cantly decreased by perillyl alcohol and sildenafil intervention.Compared with the control group,mPAP,WA(%),WT(%),right ventricular inner diameter(RVID),right ventricular free wall thickness(RVFWT)and fibrosis levels were significantly increased in the hypoxia group,while LA(%)was signifi-cantly decreased.Besides,the protein expression levels of α-SMA,ACE,AT1R and AngⅡ in lung tissues were significantly elevated while those of ACE2 and Mas,as well as Ang(1-7)content were signifi-cantly decreased in hypoxia group compared to the control group.Following intervention with perillyl alcohol and sildenafil,the protein expression levels of ACE and AngⅡin lung tissues of HPH rats were significantly decreased compared to the model group while those of ACE2 and Mas receptor,along with the content of Ang(1-7),were significantly increased.Perillyl alcohol significantly reduced the protein expression level of AT1R while sildenafil had no significant effect.CONCLUSION Perillyl alcohol can lower mPAP levels by improving pulmonary vascular remodeling and reducing pulmonary vascular fibrosis in HPH rats.The mechanism may be related to the regulatory effects on the ACE-AngⅡ-AT1R and ACE2-Ang(1-7)-Mas axes.

Relative deficiency in production of glycoprotein hormone erythropoietin (Epo) is a major cause of renal anemia. This study planned to investigate whether the hypoxia-regulated system of Epo expression, constructed by fusing Epo gene to the chimeric phosphoglycerate kinase (PGK) hypoxia response elements (HRE) in combination with cytomegalovirus immediate-early (CMV IE) basal gene promoter and delivered by plasmid intramuscular injection, might provide a long-term physiologically regulated Epo secretion expression to correct the anemia in adenine-induced uremic rats. Plasmid vectors (pHRE-Epo) were synthesized by fusing human Epo cDNA to the HRE/CMV promoter. Hypoxia-inducible activity of this promoter was evaluated first in vitro and then in vivo in healthy and uremic rats (n = 30 per group). The vectors (pCMV-Epo) in which Epo expression was directed by a constitutive CMV gene promoter served as control. ANOVA and Student's t-test were used to analyze between-group differences. A high-level expression of Epo was induced by hypoxia in vitro and in vivo. Though both pHRE-Epo and pCMV-Epo corrected anemia, the hematocrit of the pCMV-Epo-treated rats exceeded the normal (P < 0.05), but that of the pHRE-Epo-treated rats didn't. Hypoxia-regulated system of Epo gene expression constructed by fusing Epo to the HRE/CMV promoter and delivered by plasmid intramuscular injection may provide a long-term and stable Epo expression and secretion in vivo to correct the anemia in adenine-induced uremic rats.
Anemia/blood/*therapy ; Animals ; Base Sequence ; Blood Urea Nitrogen ; Cell Hypoxia ; Creatinine/blood ; Erythropoietin/biosynthesis/*genetics/secretion ; Gene Expression Regulation ; Genes, Reporter ; Genetic Therapy ; HeLa Cells ; Humans ; Injections, Intramuscular ; Kidney/pathology ; Luciferases, Firefly/biosynthesis/genetics ; Molecular Sequence Data ; Plasmids/*genetics ; Promoter Regions, Genetic ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins/biosynthesis/genetics/secretion ; Response Elements ; Transcriptional Activation ; Uremia/blood/*therapy