OBJECTIVETo analyze the expression of angiotensin II (ANG II) receptor and apoptosis in myocardium in rats of endotoxemia.

METHODSModel of endotoxemia was induced by intraperitoneal injection with lipopolysaccharide (LPS) 10 mg/kg in male Wistar rats and saline was injected into control group. The rats were killed at 2 h or 6 h after saline (control) or LPS . Expression of the correlation factors related to apoptosis of Bcl-2, Bax, AT1 and AT2 receptor in myocardial tissue were detected with immunohistochemistry (IHC), and changes of myocardial cells apoptosis was detected by the method of TUNEL. The gene expression of AT1 and AT2 receptor was examined by RT-PCR. The pathological changes of myocardial tissue were observed by electron microscope.

RESULTSCompared with control group , the expression of AT1 and AT2 receptor were significantly decreased, especially in 6 h group; and the expression of Bcl-2 and Bax were decreased, the ratio of Bcl-2/Bax had the downtrend as well as the apoptosis of myocardial cells.

CONCLUSIONInterfered by LPS, the down regulation of AT1 and AT2 receptor expression has the negative relation with apoptosis of myocardial cells, this result indicated that down regulation of AT1 and AT2 receptor expression maybe related to cardiac functional impairment, which maybe help us to find a new protective path to prevent myocardial damage induced by systemic inflammatory.

Animals ; Apoptosis ; Endotoxemia ; metabolism ; Male ; Myocytes, Cardiac ; cytology ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Receptor, Angiotensin, Type 1 ; metabolism ; Receptor, Angiotensin, Type 2 ; metabolism

Hepatitis B, Chronic ; metabolism ; Humans ; Liver Cirrhosis ; metabolism ; RNA, Messenger ; analysis ; Receptor, Angiotensin, Type 1 ; genetics ; Renin ; genetics

BACKGROUNDMany studies have suggested that angiotensin II (Ang II) and its receptors may be involved in the development of asthma. However, the expression of angiotensin II receptors (AGTR) is not clear in the lung tissue of chronic asthmatics. This study was designed to determine the relationship between airway remodeling, dysfunction and the expression of AGTRs in a rat model of asthma.

METHODSRats were sensitized with ovalbumin (OVA) for 2 weeks. Sixty minutes before an inhalation challenge, the rats were pretreated either with valsartan (15, 30, 50 mg x kg(-1) x d(-1)) or saline intragastrically. Then the rats received an OVA challenge for 30 alternative days. Acetylcholine (Ach)-induced bronchoconstriction was measured after the final antigen challenge. White cell counts in bronchoalveolar lavage fluid (BALF) and morphological changes in the airways were then assessed. The levels of transforming growth factor-beta 1 (TGF-beta(1)) and platelet-derived growth factor (PDGF) in BALF were detected by ELISA. The levels of AGTR1 and AGTR2 mRNA and protein in lung tissues were measured by RT-PCR and Western blotting.

RESULTSAGTR1 mRNA and protein levels in repeatedly OVA-challenged rats were significantly increased as compared with negative controls. The AGTR1 mRNA expression versus white cell counts of BALF and airway wall thickness (mainly in small airways) in lungs of chronic antigen-exposed rats were positively correlated. Valsartan decreased the level of AGTR1 in repeatedly OVA-challenged rats. However, AGTR2 mRNA and protein levels in the OVA-challenged rats and high-dose valsartan-treated rats (50 mg x kg(-1) x d(-1)) were also increased. Valsartan significantly decreased inflammatory cell accumulation and attenuated Ach-evoked bronchoconstriction in repeatedly antigen-challenged rats. Valsartan also decreased allergen-induced structural changes in rat airway (including total airway wall thickness and smooth muscle area) and the levels of TGF-beta(1) and PDGF in BALF.

CONCLUSIONSAGTR1 expression is potentially associated with airway remodeling and dysfunction in asthma. Ang II and AGTR1 may participate in airway inflammation and airway remodeling of chronic antigen-exposed rats. Valsartan, a AGTR1 antagonist, could inhibit AGTR1 expression and partially inhibits structural airway changes as well as airway inflammation in chronic OVA-exposed rats.

Angiotensin II Type 1 Receptor Blockers ; pharmacology ; Angiotensin Receptor Antagonists ; Animals ; Asthma ; chemically induced ; genetics ; metabolism ; Blotting, Western ; Bronchoalveolar Lavage Fluid ; chemistry ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; drug effects ; Lung ; drug effects ; metabolism ; pathology ; Male ; Ovalbumin ; Platelet-Derived Growth Factor ; metabolism ; Rats ; Rats, Wistar ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism ; Receptor, Angiotensin, Type 2 ; genetics ; metabolism ; Receptors, Angiotensin ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tetrazoles ; pharmacology ; Transforming Growth Factor beta1 ; metabolism ; Valine ; analogs & derivatives ; pharmacology ; Valsartan

OBJECTIVETo observe the effect of electroacupuncture (EA) stimulation on the expressions of angiotensinogen (AGT), angiotensin II type 1 receptor (AT1R), endothelin-1 (ET1), and endothelin A receptor (ETAR) mRNA in spontaneously hypertensive rat (SHR) aorta.

METHODSEighteen male SHRs were randomly divided into three groups, an SHR group, an SHR Baihui (DU 20) and Zusanli (ST 36) acupoint (SHR-AP) group, and an SHR non-acupoint (SHR-NAP) group, with 6 rats in each group. Six Wistar rats were used as a control. Rats in the SHR-AP group were stimulated by DU 20 and ST 36 acupoints, both of which were connected with EA. EA was handled one time every Monday, Wednesday and Friday, for total 24 times (8 weeks). SHRNAP rats were acupointed at a 15°angle flat into 0.5 cm to two points, which were 1 and 2 cm from rail tip separately. EA parameters were the same as the SHR-AP rats. SHR control rats and Wistar rats were fixed without EA. Real-time quantitative polymerase chain reaction (PCR) was used to measure AGT, AT1R, ET1, and ETAR mRNA expression in rat aorta.

RESULTSEA stimulation significantly reduced rat aorta vascular AGT, ET1, ETAR and AT1R mRNA expressions in the SHR-AP and SHR-NAP groups (P <0.01). Among these four genes, AT1R mRNA expression was significantly lower in the SHR-AP than in the SHR-NAP group (P <0.01).

CONCLUSIONEA could reduce the AT1R mRNA expression in SHR-AP rat aorta, indicating a potential mechanism for the hypotensive effects of EA.

Angiotensinogen ; genetics ; metabolism ; Animals ; Aorta ; metabolism ; physiopathology ; Blood Pressure ; Electroacupuncture ; Endothelin-1 ; genetics ; metabolism ; Gene Expression Regulation ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats, Inbred SHR ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism ; Receptor, Endothelin A ; genetics ; metabolism

Antisense Elements (Genetics) ; Cell Line ; Cell Proliferation ; Collagen Type I ; biosynthesis ; Genetic Therapy ; Hepatic Stellate Cells ; cytology ; metabolism ; Humans ; RNA, Messenger ; genetics ; Receptor, Angiotensin, Type 1 ; genetics ; Transfection

OBJECTIVETo investigate the effect of angiotensin II (AngII) type 1 (AT-1) receptor and angiotensin-converting enzyme (ACE) gene silencing on nuclear factor-kappaB (NF-kappaB) activity in hepatic stellate cells (HSCs).

METHODSpSilencer/AT-1 alpha receptor siRNA and pSilencer/ACE siRNA plasmids were transfected into cultured HSC-T6 cells, which were subsequently stimulated by 10(-6) mol/L AngII or ACE inhibitor (ACEI). The DNA binding activity of NF-kappaB in the transfected cells was analyzed using electrophoretic gel mobility shift assay (EMSA).

RESULTSs Gel shift studies showed that stimulation of the HSCs by AngII markedly increased the DNA-binding activity of NF-kappaB, which was inhibited by the transfection with pSilencer/ AT-1 alpha receptor siRNA plasmid or pSilencer/ACE siRNA plasmid.

CONCLUSIONAT-1 alpha receptor and ACE gene silencing result in inhibition of NF-kappaB activity in HSCs in vitro.

Cell Line ; Hepatic Stellate Cells ; cytology ; metabolism ; Humans ; NF-kappa B ; genetics ; metabolism ; Peptidyl-Dipeptidase A ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Receptor, Angiotensin, Type 1 ; genetics ; Transfection

This study was aimed to explore the expression level of angiotensin-II type 1 receptor (AT1) mRNA in bone marrow of myeloid leukemic patients, and its correlation with the proportion of leukemia cells in samples and Hb, WBC, Plt counting in peripheral blood. 51 samples, including 36 AML, 7 CML, and 8 samples of non-malignant hematological diseases as control group were collected. The expression of at1 mRNA was detected by real time-PCR; the expression levels of at1 gene in AML and CML groups were relatively quantitatively analyzed by using 2(-ΔΔCT) and were compared with control group. The results showed that the expression levels of at1 mRNA in AML, CML and control groups were 0.038 ± 0.076, 0.033 ± 0.039, 0.281 ± 0.366, respectively. at1 gene expression in the myeloid leukemic group was significantly lower than that in the control group. The expression level of at1 mRNA in AML was negatively correlated with the proportion of leukemia cells and positively with Hb level in peripheral blood. It is concluded that at1 gene may play a minor role in leukaemogenesis, however, may promote erythropoiesis.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Leukemia, Myeloid ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; RNA, Messenger ; genetics ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism ; Young Adult

OBJECTIVETo investigate the effect of angiotensin II type-1 (AT-1) alpha receptor gene silencing on nuclear factor-kappaB (NF-kappaB) activity in hepatic Kupffer cells.

METHODSThe expression of AT-1 alpha receptors in primary isolated cultured hepatic Kupffer cells was detected by immunohistochemistry. pSilencer/AT-1 alpha receptor siRNA plasmids were transfected into Kupffer cells, which were subsequently exposed to 10(-6) mol/L angiotensin II (Ang II) for 60 min. The changes in the DNA binding activity of NF-kappaB in the cells was assessed using electrophoretic gel mobility shift assay (EMSA).

RESULTSAT-1 alpha receptor expression was detected in Kupffer cells. NF-kappaB DNA binding activity was markedly increased in Kupffer cells after Ang II stimulation, and obviously inhibited by transfectiom with pSilencer/AT-1 alpha receptor siRNA plasmid.

CONCLUSIONAng II stimulation of Kupffer cell results in increased activation of NF-kappaB via AT-1 alpha receptor.

Cells, Cultured ; Humans ; Kupffer Cells ; cytology ; NF-kappa B ; metabolism ; RNA Interference ; RNA, Small Interfering ; genetics ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism

OBJECTIVETo investigate the effects of Shenkangwan on the expressions of angiotensin II (AngII) and its type I receptor (AT(1)R) and the renalprotection mechanism of Shenkangwan in rats with early diabetic nephropathy (DN).

METHODSThe rat models of DN established by a single injection of streptozotocin were randomly divided into 4 groups, namely the model group, Shenkangwan treatment group, irbesartan treatment group, and Shenkangwan and irbesartan treatment group, with normal rats as the control. All the rats received daily gavage for 8 weeks. The urinary protein quality in 24 h and plasma and renal contents of AngII were measured. The expressions of AT1R at the protein and mRNA levels in the kidney tissues were measured by immunohistochemistry and reverse transcription-polymerase chain reaction, respectively. The pathological changes of the kidney were observed microscopically.

RESULTSIn DN rats, Shenkangwan reduced the urinary protein quantity in 24 h and the contents of AngII in the plasma and kidney tissues, decreased the renal expressions of AT(1)R protein and mRNA, and alleviated the morphological damage of the kidney.

CONCLUSIONSShenkangwan offers renalprotection against DN probably by reducing the contents of AngII in the plasma and kidney tissues and inhibiting renal AT(1)R expressions.

Angiotensin II ; genetics ; metabolism ; Angiotensin II Type 1 Receptor Blockers ; therapeutic use ; Animals ; Diabetic Nephropathies ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Kidney ; metabolism ; Male ; Phytotherapy ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism

OBJECTIVETo study the effect of Songling Xuemaikang Capsule (SXC) on blood pressure of spontaneously hypertensive rats (SHR) and regulatory mechanisms for peroxisome proliferator activated receptor-gamma (PPARgamma).

METHODSTotally 24 10-week-old SHR rats were randomly divided into the blank control group, the Chinese medicine (CM) group, and the Western medicine (WM) group, 8 in each group. Rats in the CM group were administered with SXC at the daily dose of 20 mg/kg by gastrogavage. Those in the WM group were administered with ramipril at the daily dose of 1 mg/kg by gastrogavage. Those in the blank control group were administered with equal volume of normal saline. The blood pressure was measured once per week. The cardiac ultrasound was performed 4 weeks later. Rats were killed and then blood was sampled from abdominal aorta. mRNA expressions of liver PPARgamma and angiotensin II type 1 receptor (AT1R) were detected by fluorescence real-time quantitative PCR. Protein expressions of PPARgamma and AT1R were detected using immunohistochemical assay (SP). The contents of PPARgamma and AT1R were quantitatively analyzed by Western blot.

RESULTSAfter 4 weeks of treatment, the blood pressure decreased in the CM group, showing statistical difference when compared with the blank control group (P < 0.01). CM was inferior to WM in lowering blood pressure. But as a whole, CM was more stable and could maintain blood pressure at a relatively stable level. The cardiac ejection fraction increased in the CM group, showing statistical difference when compared with the blank control group (P < 0.05, P < 0.01). The mRNA and protein expressions of liver PPARgamma were up-regulated in the CM group, showing statistical difference when compared with the blank control group (P < 0.05, P < 0.01). CM could obviously inhibit the AT1R mRNA expression, and down-regulate the protein expression of AT1R, showing statistical difference when compared with the blank control group and the WM group respectively (P < 0.01).

CONCLUSIONSXC decreased blood pressure and improved the cardiac ejection fraction, which might be partially achieved by up-regulating the PPARgamma mRNA expression and protein synthesis, and inhibiting the AT1R mRNA expression and AT1R protein synthesis.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hypertension ; drug therapy ; metabolism ; physiopathology ; Male ; PPAR gamma ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Inbred SHR ; Receptor, Angiotensin, Type 1 ; metabolism