Mechanism of TRPC in promoting extracellular matrix deposition of glomerular mesangial cells / 西安交通大学学报(医学版)

【Objective】 To explore the role and mechanism of TRPC in promoting extracellular matrix (ECM) deposition in rat glomerular mesangial cells (HBZY-1). Methods Immunofluorescence staining was performed to observe the distribution and expression of TRPC1 and TRPC6 in HBZY-1 cells. After AngⅡ stimulation, qRT-PCR and Western blotting were used to detect the mRNA and protein expressions of Gαq/PLCβ4/TRPC signaling pathway main proteins and ECM deposition indicators (α-SMA, collagenⅢ and fibronectin). By silencing the expressions of TRPC1 and TRPC6 by RNA interference, the expressions of ECM deposition indicators were detected. Changes in [Ca2+]i influx were determined through Fluo-4AM Ca2+ imaging. 【Results】 Both TRPC1 and TRPC6 were expressed in HBZY-1, and were mainly located in cell membrane and cytoplasm. After AngⅡ stimulation, Gαq/PLCβ4/TRPC signaling pathway was activated, and the mRNA and protein expressions of Gαq, PLCβ4, TRPC1 and TRPC6 were all increased (P<0.05). [Ca2+]i influx also increased (P<0.01), and the mRNA and protein expressions of ECM deposition indicators (α-SMA, ColⅢ and Fn) were upregulated (P<0.05). Silencing the expressions of TRPC1 and TRPC6 by RNA interference led to decreased [Ca2+]i influx (P<0.05), and downregulated mRNA and protein expressions of ECM deposition indicators in HBZY-1 cells (P<0.05). The results suggested that inhibition of TRPC expressions could inhibit AngⅡ induced ECM deposition in HBZY-1 cells, which might be associated with decreased [Ca2+]i influx. 【Conclusion】 TRPC may be a novel therapeutic target of renal fibrosis.

Effect and mechanism of leonurine on pressure overload-induced cardiac hypertrophy in rats / 中国中药杂志

To investigate the effects of leonurine(Leo) on abdominal aortic constriction(AAC)-induced cardiac hypertrophy in rats and its mechanism. A rat model of pressure overload-induced cardiac hypertrophy was established by AAC method. After 27-d intervention with high-dose(30 mg·kg~(-1)) and low-dose(15 mg·kg~(-1)) Leo or positive control drug losartan(5 mg·kg~(-1)), the cardiac function was evaluated by hemodynamic method, followed by the recording of left ventricular systolic pressure(LVSP), left ventricular end-diastolic pressure(LVESP), as well as the maximum rate of increase and decrease in left ventricular pressure(±dp/dt_(max)). The degree of left ventricular hypertrophy was assessed based on heart weight index(HWI) and left ventricular mass index(LVWI). Myocardial tissue changes and the myocardial cell diameter(MD) were measured after hematoxylin-eosin(HE) staining. The contents of angiotensin Ⅱ(AngⅡ) and angiotensin Ⅱ type 1 receptor(AT1 R) in myocardial tissue were detected by ELISA. The level of Ca~(2+) in myocardial tissue was determined by colorimetry. The protein expression levels of phospholipase C(PLC), inositol triphosphate(IP3), AngⅡ, and AT1 R were assayed by Western blot. Real-time quantitative PCR(qRT-PCR) was employed to determine the mRNA expression levels of β-myosin heavy chain(β-MHC), atrial natriuretic factor(ANF), AngⅡ, and AT1 R. Compared with the model group, Leo decreased the LVSP, LVEDP, HWI, LVWI and MD values, but increased ±dp/dt_(max) of the left ventricle. Meanwhile, it improved the pathological morphology of myocardial tissue, reduced cardiac hypertrophy, edema, and inflammatory cell infiltration, decreased the protein expression levels of PLC, IP3, AngⅡ, AT1 R, as well as the mRNA expression levels of β-MHC, ANF, AngⅡ, AT1 R, c-fos, and c-Myc in myocardial tissue. Leo inhibited AAC-induced cardiac hypertrophy possibly by influencing the RAS system.

Effect of Fushengong Prescreption on Regulation-antagonism Effect of ACE-AngⅡ-AT1R Axis and ACE2-Ang(1-7)-MASR Axis of Rats with Chronic Renal Failure / 中国实验方剂学杂志

Objective:To study the effect of Fushengong prescreption on the regulation-antagonism effect of angiotensin converting enzyme-angiotensin Ⅱ-angiotensin Ⅱ 1 receptor （ACE-AngⅡ-AT1R） axis and angiotensin converting enzyme 2-angiotensin （1-7）-Mas receptor［ACE2-Ang（1-7）-MASR］ axis of rats with chronic renal failure（CRF）， and to explore its mechanism of delaying the development of CRF. Method:The 65 male SD rats were randomly divided into normal group （<italic>n</italic>=10） and modeling group （<italic>n</italic>=55）. The normal group was routinely reared， while the modeling group were administered by gavage with 0.25 g·kg^-1d^-1adenine suspension for 28 days. After the model was successfully established， the survival model rats were randomly divided into model group， benazepril group（0.01 g·kg^-1·d^-1）and low，medium and high dose of Fushengong prescreption groups （4，8，16 g·kg^-1·d^-1）. The normal group and model group were administered the same volume of normal saline by gavage， lasted for 28 days. After the experiment， systolic blood pressure （SBP） and diastolic blood pressure （DBP） of caudal artery were measured， and 24-hour urine was collected to determine 24-hour urine protein （24 h U-pro）. The content of serum creatinine（SCr） and blood urea nitrogen （BUN） in the serum were measured， the histological morphology was observed by hematoxylin eosin（HE）staining， and the degree of renal interstitial fibrosis was observed by Masson staining. Enzyme linked immunosorbent assay （ELISA） was used to determine the contents of AngⅡ， Ang （1-7） and Cystatin C （CysC） in serum and renal homogenate. The protein level of ACE， ACE2， AT1R and MASR were detected by Western blot. The expression of ACE and ACE2 protein in renal tissues were detected by immunohistochemistry. Result:Compared with normal group， the expression levels of SCr， BUN and CysC in model group were significantly increased（<italic>P</italic><0.05）， the content of AngⅡ in serum and kidney tissues were significantly increased， the content of Ang （1-7） were significantly decreased（<italic>P</italic><0.05）， the expression of ACE and AT1R protein in renal tissues were significantly increased（<italic>P</italic><0.05）， and the expression of ACE2 and MASR protein were significantly decreased（<italic>P</italic><0.05）. Compared with model group and benazepril group， after the intervention with Fushengong prescreption， the serum SCr，BUN and CysC decreased（<italic>P</italic><0.05），the content of AngⅡ in serum and kidney tissues decreased significantly，Ang（1-7） increased significantly（<italic>P</italic><0.05）， the expression of ACE and AT1R protein in renal tissues decreased significantly（<italic>P</italic><0.05）， ACE2 and MASR protein increased significantly（<italic>P</italic><0.05）. The high-dose Fushengong prescreption has the best effect. The high， medium and low-dose effects of Fushengong prescreption were dose-dependent. Conclusion:Fushengong prescreption improved renal function and pathological change of kidney in adenine-induced rats with chronic renal failure. The mechanism may be related to the inhibition of ACE-AngⅡ-AT1R axis and promotion of ACE2-Ang（1-7）-MASR axis ，which leads to the delaying of the progression of chronic renal failure.

Protective Mechanism of Danggui Shaoyaosan on Podocytes of Nephrotic Syndrome Rats Based on AngⅡ-TRPC6 Pathway / 中国实验方剂学杂志

Objective:To explore the protective effect and the mechanism of Danggui Shaoyaosan（DSS） on angiotensin Ⅱ （AngⅡ）/transient receptor potential cation channel 6 （TRPC6） pathway in nephrotic syndrome （NS） rats. Method:In animal experiments， doxorubicin （4 mg·kg^-1 for the 1^st week and 2 mg·kg^-1 for the 2^nd week） was injected twice to the tail vein of rats to induce NS model in 160 rats， which were then randomly divided into model group （normal saline）， losartan group （30 mg·kg^-1·d^-1）， and low-（4.3 g·kg^-1·d^-1）， medium-（8.6 g·kg^-1·d^-1）， and high-dose （17.2 g·kg^-1·d^-1） DSS groups. Besides， a normal group was also set. After intervention for four weeks， ultrastructure changes of the kidney were identified by transmission electron microscopy （TEM）. The 24-hour urine protein was detected by kits. Radioimmunoassay was used to detect the content of AngⅡ and Calcineurin （CaN） in plasma. Western blot was used to detect the protein expression of TRPC6， angiotensin Ⅱ type 1 receptor （AT1R）， podocyte slit diaphragm-specific protein （Nephrin）， and cysteine-aspartic acid protease-3 （Caspase-3） in the renal cortex. Immunohistochemistry was used to detect the expression of TRPC6 and AT1R in the slit diaphragm. In cell experiments， AngⅡ stimulated MPC5 podocytes. The cells were randomly divided into a normal group， an AngⅡ group， an AngⅡ+SAR7334 （TRPC6-specific inhibitor） group， an AngⅡ+5%DSS group， an AngⅡ+10%DSS group， and an AngⅡ+15%DSS group. Western blot was used to detect the protein expression of TRPC6， AT1R， Nephrin， and Caspase-3 in podocytes. Result:Compared with the normal group， the model group showed increased 24-hour urine protein content （<italic>P</italic><0.01） and AngⅡ and CaN in plasma （<italic>P</italic><0.01）， incomplete glomerular structure， the extensive fusion of podocyte process with elevated fusion rate （<italic>P</italic><0.01）， increased expression distribution of AT1R and TRPC6 in the renal cortex， and up-regulated protein expression of AT1R， TRPC6， and Caspase-3 in renal tissues （<italic>P</italic><0.01）， and reduced Nephrin protein expression （<italic>P</italic><0.01）. Compared with model group， the losartan group and the high-dose DSS group exhibited decreased 24-hour urine protein content （<italic>P</italic><0.01） and the content of AngⅡ and CaN in plasma （<italic>P</italic><0.01）， improved glomerular structure， reduced fusion rate of podocyte process （<italic>P</italic><0.01）， diminished expression distribution of TRPC6 and AT1R in the renal cortex， declining protein expression of AT1R， TRPC6 and Caspase-3 in renal tissues （<italic>P</italic><0.01）， and elevated Nephrin protein expression （<italic>P</italic><0.01）. Additionally， compared with the normal podocytes， AngⅡ-stimulated podocytes showed increased protein expression of AT1R， TRPC6 and Caspase-3 （<italic>P</italic><0.01）， and decreased expression of Nephrin （<italic>P</italic><0.01）. Compared with the AngⅡ group， the AngⅡ+SAR7334 group displayed reduced protein expression of AT1R， TRPC6， and Caspase-3 （<italic>P</italic><0.01） and increased protein expression of Nephrin （<italic>P</italic><0.01）. Conclusion:DSS can improve the pathological characteristics of NS presumedly by inhibiting the interaction between AngⅡ and TRPC6 in podocytes and improving the structural integrity of podocytes to repair the damage of glomerular molecular barrier and slow down the progression of NS-induced proteinuria.

Effects of notoginsenoside R1 on AngⅡ induced proliferation of MOVAS cells and AT1R/MAPKs signaling pathway / 上海预防医学

Objective:To investigate the effects of notoginsenoside R1 （NR1） on the proliferation of mice aortic smooth muscle cells （MOVAS cells） induced by angiotensinⅡ （AngⅡ） and the signal pathway of angiotensin Ⅱ type 1 receptor （AT1R） / mitogen activated protein kinases （MAPKs）. Methods:The proliferation of MOVAS cells was detected by BrdU method after AngⅡ induction. Western blot was used to detect the expression of the two main receptors of AngⅡ （AT1R and AT2R） and MAPKs pathway related proteins （ERK， p38， and JNK）. Results:（1） AngⅡ （5 μmol/L） could promote the proliferation of MOVAS cells （P<0.01）. NR1 （50 μmol/L） could inhibit the proliferation of MOVAS cells induced by AngⅡ （P<0.01）. There was no significant difference between control group and NR1 group （P>0.05）. （2） Compared with AngⅡ group， the expression of AT1R protein in AngⅡ+ NR1 group was significantly lower （P<0.05）， but there was no difference in the expression of AT2R protein （P>0.05）. （3） NR1 could significantly inhibit the phosphorylation of ERK， p38 and JNK protein after AngⅡ stimulation （P<0.01）. Conclusion:NR1 can inhibit the proliferation of MOVAS cells induced by AngⅡ， which may be related to down regulating AT1R and inhibiting the activation of MAPKs.

Effect of Kangxianling Decoction on Renal Fibrosis and ACE-AngⅡ-AT1R Axis in 5/6 Nephrectomized Rats / 中国实验方剂学杂志

Objective: To study the therapeutic effect of Kangxianling decoction on renal fibrosis induced by 5/6 nephrectomy, and angiotensin converting enzyme-angiotensin Ⅱ-angiotensin Ⅱ 1 receptor (ACE-AngⅡ-AT1R) axis. Method: Totally 50 SD rats were randomly divided into the following groups:control group (n=10), sham-operation group (n=10), 5/6 nephrectomized renal fibrosis model group (n=30). After two weeks, the rats in operation group were divided into the model group, Kangxianling group, and losartan potassium group, n=10 in each group. Rats in losartan potassium group were administered with losartan potassium by gastrogavage, and rats in Kangxianling group were administered with Kangxianling by gastrogavage. Equal volume of saline was administered to rats in the other groups. The rats were put to death after 16 weeks of consecutive medication, and serum creatinine(SCr), blood urea nitrogen(BUN), 24 h urine protein(24 h-Pro) were measured in each group. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of kidney tissues, and the degree of renal fibrosis was observed by Masson staining. The expressions of ACE1 and AT1R were detected by immunohistochemistry. The protein expression levels of ACE1, AngⅡ and AT1R were determined by Western blot. Result: Compared with control and sham-operation groups, SCr, BUN and 24 h-Pro in model group were significantly increased (PPPPConclusion: Kangxianling decoction can delay the progress of renal fibrosis in 5/6 nephrectomized rats, which is closely related to the inhibition of ACE-AngⅡ-AT1R axis activation.

Effect of ACE/Ang Ⅱ/AT1 axis and ACE2/Ang(1-7)/Mas axis on glucose and lipid metabolism in adipose tissue / 基础医学与临床

The renin angiotensin system(RAS)includes two counterbalance axes:the ACE/Ang Ⅱ/AT1 axis and the ACE2/Ang(1-7)/Mas axis.The RAS can regulate glucose and lipid metabolism in adipose tissue.Most of evi-dences demonstrated that the ACE/Ang Ⅱ/AT1 axis can induce glucose metabolism disorders in adipose tissue, while ACE2/Ang(1-7)/Mas axis improves glucose metabolism.The RAS,which is over activated in obese patient, has been considered to be a potential link among obesity,dyslipidemia and insulin resistance.The effect of ACE/AngⅡ/AT1 axis and ACE2/Ang(1-7)/Mas axis on lipid and glucose metabolism in adipose tissue should be futh-er investigated,and we may find a new target for improving glucose and lipid metabolism.

Use BAPN and AngⅡ to establish SD rat aortic dissection model / 中华胸心血管外科杂志

Objective To establish the SD rat aortic dissection(AD)model by using both BAPN and AngⅡ,in order to investigate AD's pathogenesis. Methods 90 three weeks old SD rats were equally divided into three groups randomly:control group,medicine gavage group and blank medicine gavage group. Rats in control group were fed on a regular diet;BAPN ( 1g/ kg per day)was forced into rats'stomach in the medicine gavage group;the same volume saline was forced into rats' stomach in the blank medicine gavage group. 4 weeks later,when the rats were 7 weeks old,we stopped giving them BAPN, but to implant an omicro-osmotic pump subcutaneously in the abdomen. The pumps in control group and blank medicine gavage group were filled with 0. 9％ saline,the medicine gavage group'pumps were filled with AngⅡsolution( 1 μg·kg- 1 ·min- 1 ). 1 week later,all the survivals were dissected after anesthesia and the aortic vessels were acquired. All the acquired aortic ves-sels were proceed pathological examination. All the rats dead during the process of the experiment were dissected immediately to get the aortic vessels and proceed pathological examination. Results All rats in control group and blank medicine gavage group were survival,there was no aortic dissection or death. In medicine gavage group, 15 rats developped aortic dissection, 12 a-mong them were died of aortic dissection rupture,the aortic dissection formation rate was 50％ . Conclusion Using BAPN and AngⅡ to establish the SD rat AD model is feasible,it is simple and practicable,meanwhile,it has high aortic dissection for-mation rate. The process is similar with human's aortic dissection process.

Effects of electroacupuncture on postoperative cognitive dysfunction and AngⅡ/AT1R in the hippocampus in aging rats induced by D-galactose / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of electroacupuncture (EA) on postoperative cognitive dysfunction (POCD) and AngⅡ/AT1R in the hippocampus in D-galactose-induced aging rats which received hepalobectomy, and to explore the possible mechanism of EA on POCD.</p><p><b>METHODS</b>Eighty male Sprague-Dawley rats were randomly divided into a young control group (10 rats), a D-Galactose-induced aged (Da) group (10 rats), a Da+hepatolobectomy group (30 rats) and an EA group (30 rats). The rats in the Da+hepatolobectomy group and EA group were further randomly divided into a 1 d subgroup, 3 d subgroup and a 7 d subgroup, 10 rats in each subgroup. The rats in the EA group were treated with EA at "Baihui" (GV 20) and "Dazhui" (GV 14) with continuous wave (15 Hz in frequency and 1 mA in intensity), and rats in each subgroup were treated for 1 d, 3 d and 7 d, respectively. The rats in the remaining groups were treated with immobilization, once a day. The Y-maze was used to observe the behavior change of rats, and ELISA was applied to measure the level of hippocampal AngⅡ, and RT-PCR and immunohistochemistry method were performed to detect AT1R mRNA expressions and AT1R positive expression in the hippocampus.</p><p><b>RESULTS</b>The number of rat initiative avoidance in the Da group was significantly less than that in the young control group (<0.05), and the mRNA expression and positive percentage of AT1R in the hippocampus in the Da group were significantly higher than those in the young control group (both<0.01). Compared with the Da group, the number of rat initiative avoidance in each subgroup of Da+hepatolobectomy group and EA group were significantly reduced (all<0.01), and the expression of AngⅡ, AT1R mRNA and AT1R positive cells percentage in the hippocampus were significantly increased (<0.05,<0.01). The number of rat initiative avoidance in each subgroup of EA group was higher than that in the subgroup of Da+hepatolobectomy group (<0.05,<0.01); and the expression of AngⅡ, AT1R mRNA, and AT1R positive percentage in the EA group were significantly less than that in the Da+hepatolobectomy group (<0.05,<0.01).</p><p><b>CONCLUSIONS</b>EA at "Baihui" (GV 20) and "Dazhui" (GV 14) could improve POCD in D-galactose-induced aging rats which received hepalobectomy, and it is likely to be related with the inhibition of AngⅡ, AT1R positive expression and AT1R mRNA in the hippocampus.</p>

Effect of Anti-Renin Angiotensin System Drugs in Tumors / 医药导报

Initially,the renin-angiotensin system (RAS) was considered to play an important role in regulating cardiovascular function and maintaining the balance of water and electrolyte.Based on this,several targeted drugs in the treatment of hypertension were developed.With the large-scale clinical apphcation of these drugs,RAS inhibitors are found to has a significant inhibitory effect on some of the tumor development,which reveals the RAS function in cell proliferation,differentiation,angiogenesis and tumor occurrence.In this paper,the important physiological ftmction of RAS in tumor occurrence and development were reviewed.

Antihypertensive Effect of the Extract from Compound Prunella Vulgaris L. in Spontaneous Hypertension Rats / 中国药师

Objective:To study the antihypertensive effect of the extract from compound Prunella vulgaris L. in spontaneous hy-pertension(SH)rats. Methods:Forty SH rats were randomly divided into the model group,high,middle and low dose groups of ex-tract from vompound Prunella vulgaris L. ,the compound kendir leaves tablets Ⅰ group with 8 ones in each. Non-invasive blood pres-sure measurement was used to detect the SBP and DBP of the SHR rats. Then the serum NO,AngⅡ ,ET-1 and ANP content were measured after the eight-week treatment. The pathological changes were observed after kidney HE staining in the SH rats. Results:Compared with that in the model group,the blood pressure in high-dose treatment group,middle-dose treatment group and the positive model group was significantly decreased(P < 0. 05). The AngⅡ and ET-1 levels in high-dose treatment group,middle-dose treatment group and the positive model group were decreased(P < 0. 01),and NO and ANP contents in serum were significantly increased when compared with those in the model group(P < 0. 01). The pathological examination showed that the pathological changes in the model group were faster than those in all the drug-treatment groups,the pathological changes included glomerular and renal tubular atrophy, glomerular vascular wall thickening and renal tubular epithelial cell degeneration or necrosis. Conclusion:The extract from compound Prunella vulgaris L. can reduce blood pressure of SH rats. The mechanism may be associated with the level reduction of AngII and ET-1 and content elevation of NO and ANP.

Curcumin inhibits AngⅡ-induced proliferation and oxidative stress in vascular smooth muscle cells / 西安交通大学学报(医学版)

Objective To investigate the effect of curcumin (Cur)on AngⅡ-induced proliferation and oxidative stress of vascular smooth muscle cells (VSMCs).Methods Primary rat VSMCs were cultured and divided into control group,AngⅡ group,AngⅡ+Cur 5μmol/L group,AngⅡ+Cur 10μmol/L group,AngⅡ+Cur 20μmol/L group,and Cur 20μmol/L group.The proliferation of AngⅡ-induced VSMCs was measured by MTT assay.The mRNA and protein expressions of inducible nitric oxide synthase (iNOS)and p47phox were detected by real-time PCR and Western blot.Nitric oxide (NO)production was measured by Griess reaction.Production of intracellular reactive oxygen species (ROS)was measured by DCFH-DA staining,and the activities of superoxide dismutase (SOD)and glutathione peroxidase (Gpx)were detected by xanthine oxidase assay and visible spectrophotometer. small interfering RNA (siRNA)was used to silence the expression of p47phox to further explore the mechanism for Cur inhibiting the proliferation of AngⅡ-induced VSMCs and oxidative stress.Results VSMCs activities were not significantly affected by Cur at the concentration between 0 and 80μmol/L.Cur (5,10 and 20μmol/L)significantly inhibited AngⅡ-induced proliferation of VSMCs.Cur had an inhibitory effect on the overexpression of NO,iNOS, p47phox and ROS in VSMCs and upregulated the activities of SOD and Gpx in a concentration-dependent manner. AngⅡ-induced ROS production in VSMCs was significantly attenuated by pretreatment with p47phox specific siRNA.Conclusion Cur can inhibit the proliferation and oxidative stress of AngⅡ-induced VSMCs.

Correlation between the serum concentration of ACE2/Ang (1-7)and the occurrence of atrial fibrillation in patients with rheumatic valvular heart disease / 西安交通大学学报(医学版)

Objective To reveal the role of serum ACE2/Ang (1-7)in the occurrence of atrial fibrillation (AF)and find new targets for the prevention and treatment of AF by analyzing the correlation between the serum concentration of ACE2/Ang (1-7 )in patients with rheumatic valvular heart disease and the occurrence of AF. Methods We collected the basic clinical information and peripheral venous blood of patients with rheumatic heart valve disease (totally 46 patients,including 24 with AF and 22 with SR).ELISA method was used to detect the serum concentration of ACE2,Ang (1-7)and AngⅡ in the serum samples.Then the differences and correlation between the two groups were analyzed.Results In the AF group ① the diameter of the left atrium was significantly greater than that in the SR group [(60.70±3.08 vs.48.15±2.16)mm,P<0.05];② the serum concentration of AngⅡ was significantly higher than that in the SR group [(45.88±2.87 vs.35.78±1.08)pg/mL, P<0.05],AngⅡ and left atrium diameter were positively correlated (Pearson test,P<0.05);③ the serum concentrations of ACE2 [(7.87±0.74 vs.11.65±0.57)U/L,P<0.05]and Ang (1-7)[(146.05±17.61 vs. 321.71±36.50)pg/mL,P<0.05]were significantly lower than those in the SR group,and negatively correlated with left atrium diameter (Pearson test,P<0.05);④ the serum concentration of Ang (1-7)was negatively correlated with AngⅡ concentration (Pearson test,P<0.05).Conclusion For patients with rheumatic valvular heart disease,ACE2/Ang (1-7 )may play a protective role in the occurrence of AF via antagonizing AngⅡ and inhibiting atrial remodeling.

AngⅡ promotes liver fibrosis through PDGF signal pathway / 实用医学杂志

Objective To investigate the effect and mechanism of AngⅡ on collagen in hepatic stellate cell. Methods HSCs were isolated and cultured, 3H-pro incorporation method was used to evaluate the effects of different doses of AngⅡ on the proline syntheses. RT-PCR assay were used to assess changes in mRNA expression levels of type Ⅰ and Ⅲ procollagen. PDGFR-β mRNA and protein were determined by in situ hybridization and immunocytochemistry. Results 10-8~ 10-5 mol/L AngⅡ could significantly increase the 3H-pro incorporation rate of HSC in a dose-dependent style, 10-6 mol/L AngⅡis the most effective dose. The cultured HSC showed a little expression of type Ⅰ and Ⅲ procollagen mRNAs, while 10-6 mol/L AngⅡwas able to enhance the expression for type Ⅰ and Ⅲ procollagen mRNAs significantly(P < 0.01). AngⅡalso could enhance both mRNA and protein expression of PDGFR-β on HSC(P < 0.01). Conclusion These results suggest that AngⅡ could promote HSC collagen synthesis by enhancing the expressions of PDGFR-β.

Effect of ShenQi FuZheng injection on peripheral blood AngⅡ, ET-1 levels and clinical curative effect in patients with congestive heart failure / 中国生化药物杂志

Objective To analysis effect of ShenQi FuZheng injection on peripheral blood angiotensin II(AngⅡ), endothelin-1(ET-1) levels and clinical curative effect in patients with congestive heart failure.Methods 52 patients who were diagnosed with chronic heart failure were collected.All patients were randomly divided into experimental group and control group, 26 cases in each group.Patients in the control group received conventional western medicine treatment, patients in the experimental group were given ShenQi FuZheng injection on the basis of control group treatment, after the treatment, the plasma levels of AngⅡ and ET-1, cardiac function and clinical efficacy were detected in all patients.Results After treatment, compared with control group, the plasma levels of AngⅡ was lower in the experimental group(P<0.05);the plasma levels of ET-1 was lower in the experimental group(P<0.05); the LVEF,CO and SV levels were higher,and the LVED level was lower in the experimental group(P<0.05); the effective rate of patients in the experimental group was higher(P<0.05).Conclusion The ShenQi FuZheng injection can significantly reduce the plasma AngⅡand ET-1 levels in patients with congestive heart failure, improve heart function and clinical curative effect.

Effect ofXiao-Qing-LongDecoction on AngⅡ, ALD, Na+-k+-ATPase Contents and Expressions of AT1 and AT2 mRNA in Cor pulmonale Rats / 世界科学技术-中医药现代化

This article was aimed to study the effect ofXiao-Qing-LongDecoction (XQLD) on plasma AngⅡ, ALD, Na+-k+-ATPase content, and plasma AT1 and AT2 mRNA expressions in Cor pulmonale rats, in order to further explore the mechanism of ventilating lung qi for diuresis. A total of 60 Wistar rats were randomly divided into the normal group, model group and XQLD, with 20 rats in each group. The enzyme-linked immunosorbent assay (ELISA) method was used to determine the AngⅡ, ALD and Na+-k+-ATPase content. The fluorescence quantitative PCR was used to detect the AT1 and AT2 mRNA expression. The results showed that compared with the normal group, the AngⅡ, ALD and Na+-k+-ATPase contents in the model group were significantly increased (P< 0.05, orP< 0.01). Compared with the model group, the AngⅡ, ALD and Na+-k+-ATPase contents in the XQLD group were obviously decreased (P< 0.05). Compared with the normal group, AT1 mRNA expression was increased; and AT2 mRNA expression was decreased in the model group (P<0.05, orP< 0.01). Compared with the model group, AT1 mRNA expression was decreased; and AT2 mRNA expression was increased in the XQLD group (P < 0.01). It was concluded that XQLD can effectively regulate the AT1 and AT2 mRNA expressions, influence ALD content to ventilate lungqi for dieresis.

AngⅡin paraventricular nucleus contribute to chronic intermittent hypoxia induced-hypertension in rats / 中国药理学通报

Aim To determine whether AngⅡin para-ventricular nucleus (PVN)was involved in the chronic intermittent hypoxia (CIH ) induced-hypertension in rats.Methods Male Sprague-Dawley rats were ran-domly divided into Sham and CIH groups,the Sham rats were exposed to continuous normoxia,while the CIH rats were submitted to CIH (8 h per day for 15 days).The conscious noninvasive method with tail cuff was performed in rats to record the systolic blood pres-sure during establishing the model of CIH induced hy-pertension.Mean arterial pressure (MAP)and heart rate (HR)were recorded in vivo on a PowerLab data acquisition system after CIH.Rats were fixed on the stereotaxic instrument to conduct microinjection in the PVN.We used Western blot to measure Ang Ⅱ level and AngⅡtype 1 receptor (AT1 R)protein expression in PVN.Results The level of PVN Ang Ⅱin CIH rats was significantly higher than that in Sham rats,a-long with increased AT1 R protein expression.Microin-jection of Ang Ⅱ(0.03,0.3,3 nmol)in bilateral PVN dose-dependently increased MAP in both CIH and Sham rats,and this response was significantly augmen-ted in CIH rats.Losartan (50 nmol),AT1 R antago-nist,had no effect on MAP in Sham rats,but caused significant MAP decreases in CIH rats,and prevented Ang Ⅱ-induced increases in MAP in both CIH and Sham rats.Conclusion The results suggest that the increased AngⅡrelease and enhanced AT1 R activation in the PVN contribute to CIH induced-hypertension in rats.

Effect of AngⅡ on Insulin Gene Expression in RIN-m Cells and Its Molecular Mechanism / 中国药师

Objective:To discuss the effect of AngⅡ on insulin gene expression IN RIN-m cells and its molecular mechanism. Methods:RIN-m cells were cultured and divided into three groups, including the control group, 100 nmol·L-1 AngⅡ group and losartan pretreatment group. After 24-hour incubation, insulin gene expression in RIN-m cells was detected by RT-PCR, the mean flu-orescent intensity of 2', 7'-dichlorofluorescein ( DCF) was detected by flow cytometry, PDX-1 and MafA mRNA expression were detec-ted by RT-PCR and the protein expression was detected by Western-blot. Results: Insulin expression in RIN-m cells, cellular ROS level, PDX-1 expression and MafA expression in 100 nmol · L-1 AngⅡ group were significantly different from those in the control group and losartan pretreatment group (P0. 05). Conclusion:AngⅡ can down-regulate PDX-1 and MafA expression inβ-cells through oxidative stress pathway, and then inhibit insu-lin gene expression. Pretreatment with losartan can antagonize the effect of AngⅡ, and has protective effect onβ-cells in the aspect of insulin gene expression.

Mechanisms ofJia-ShenPrescription on AngⅡ Inhibition and Ventricular Remodeling in Myocardial Infarction Rat Model / 世界科学技术-中医药现代化

This study was aimed to observe the effect ofJia-Shen prescription (JSP) on angiotensinⅡ (AngⅡ) inhibition, ventricular remodeling in myocardial infarction (MI) rat model. The anterior descending coronary artery of Sprague-Dawley rat was ligated to establish the MI rat model. Rats were randomly divided into the 3 g JSP group, 6 g JSP group, losartan group, model group, and the sham-operation group. Intragastric administration of medication was given 24 h after MI. In the 3 g and 6 g JSP group, JSP was given at the dose of 3 g·kg-1·day-1 and 6 g·kg-1day-1, respectively. Losartan was given at the dose of 10 mg·kg-1·day-1 in the losartan group. Same volume of distilled water was given to the sham-operation and model group. Four weeks later, the left ventricular end-systolic diameter (LVESD), left ventricular end-diastolic diameter (LVEDD), posterior wall thickness (PWT), left ventricular ejection fraction (LVEF), left ventricular fractional shorten (LVFS), left ventricular weight index (LVWI), the distribution and content of collagen, plasma brain natriuretic peptide (BNP) and the AngⅡ content in myocardial tissues homogenate were observed. The results showed that 4 weeks after MI, compared to the model group, 6 g PJP reduced myocardial infarct size, LVWI, LVEDD and LVESD, and enhanced LVEF and LVFS (P< 0.05). In ischemic regions, compared to the model group, JSP can obviously reduce the content of collagen (P < 0.05). This effect had dose-dependent relationship. Plasma BNP and AngⅡ content in myocardial tissues homogenate were also obviously lower than the model group (P< 0.05). It was concluded that JSP can improve the ventricular remodeling of MI rat model. Its action mechanism may be through the AngⅡ inhibition, in order to improve the early stage left ventricular morphological remodeling, myocardial fibrosis and cardiac contractile function.

Advances in Proliferation Model of Vascular Smooth Muscle Cells and Inhibitory Effects and Mechanisms of Chinese Herbal Medicine / 世界科学技术-中医药现代化

The proliferation of vascular smooth muscle cells is closely related to the pathogenesis of a variety of diseases, such as atherosclerosis, hypertension, diabetes, vascular restenosis, and etc. A proper model for smooth muscle cell proliferation can be served as an important tool for exploring the molecular mechanisms of smooth muscle cell proliferation and the screening of potential inhibitory agents. Various factors can induce the proliferation of smooth muscle cells with different mechanisms and pathological significance. The ox-LDL, AGEs, angiotensin II (AngⅡ), high glucose, tumor necrosis factor (TNF) and etc. have been successfully applied in the establishment of smooth muscle cell proliferation model. This article summarized the most commonly used models for smooth muscle cell proliferation. The inhibitory effects of Chinese herbal medicine, especially the isolated pure compounds were discussed. It was aimed to provide references for the screening of smooth muscle cells based medications as well as studies on related research and development of Chinese herbal medicine.