Modified Liuwei Dihuangtang Combined with Losartan Potassium Regulates ACE1/AngⅡ/AT1R Axis and Intestinal Flora in Rat Model of Diabetic Kidney Disease / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of modified Liuwei Dihuangtang in preventing and treating renal injury in diabetic kidney disease （DKD） via the angiotensin-converting enzyme 1 （ACE1）/angiotensin Ⅱ （AngⅡ）/angiotensin Ⅱ type 1 receptor （AT1R） axis. MethodFifty male SD rats were randomized into a normal group （n=8） and a modeling group （n=42）. The rats in the modeling group were fed with a high-sugar and high-fat diet for 6 weeks and intraperitoneally injected with 35 mg·kg-1 streptozotocin （STZ） to establish the model of DKD. After successful modeling， the rats were randomized into model， traditional Chinese medicine （modified Liuwei Dihuangtang granules 21 g·kg-1）， western medicine （losartan potassium， 33 mg·kg-1）， and integrated Chinese and western medicine （losartan potassium 33 mg·kg-1 combined with modified Liuwei Dihuangtang granules 21 g·kg-1） groups. The levels of fasting blood glucose （FBG）， urinary protein （Up）， blood urea nitrogen （Bun）， and serum creatinine （SCr） were measured in each group after 8 consecutive weeks of drug intervention. Enzyme-linked immunosorbent assay was employed to determine the serum levels of ACE1， AngⅡ， and AT1R. Western blot was employed to measure the protein levels of ACE1， AngⅡ， and AT1R in the renal tissue. The pathological and morphological changes of the renal tissue were observed after hematoxylin-eosin （HE） staining， Masson staining， and periodic acid Schiff 's （PAS） staining. The fecal samples of rats in each group were collected for 16S rDNA high-throughput sequencing. ResultCompared with the normal group， the model group showed elevated levels of Up， FBG， Bun， SCr， ACE1， AngⅡ， and AT1R （P<0.01）， serious lesions in the renal tissue， up-regulated protein levels of ACE1， AngⅡ， and AT1R （P<0.01）， increased Firmicutes/Bacteroidetes （F/B） ratio， decreased relative abundance of Lactobacillus， and increased relative abundance of Moralella and Bifidobacteria. Compared with the model group， drug intervention lowered the levels of Bun， SCr， ACE1， AngⅡ， and AT1R （P<0.01） and alleviated the pathological changes in the renal tissue. Chinese medicine and integrated Chinese and western medicine lowered the levels of Up and FBG （P<0.01）， and western medicine and integrated Chinese and western medicine down-regulated the protein levels of ACE1， AngⅡ， and AT1R. In addition， Chinese medicine down-regulated the protein levels of AngⅡ （P<0.01） as well as ACE1 and AT1R （P<0.05）. Chinese medicine and integrated Chinese and western medicine decreased the F/B ratio， and western medicine and Chinese medicine increased the relative abundance of Blautia. Chinese medicine and integrated Chinese and western medicine increased the relative abundance of Lactobacillus， Ruminococcus undetermined genera， and Bifidobacteria， decreased the relative abundance of Moralella， and increased the Chao 1 and Ace indexes （P<0.05）. Compared with the western medicine group， the integrated Chinese and western medicine group showed lowered levels of Up （P<0.01）， Bun （P<0.05）， and ACE1 and AT1R （P<0.01）， down-regulated protein levels of ACE1， AngⅡ， and AT1R （P<0.05）， alleviated pathological changes in the renal tissue， increased relative abundance of Bifidobacteria， and increased Chao 1 and Ace indexes （P<0.05）. ConclusionModified Liuwei Dihuangtang combined with losartan potassium can mitigate renal fibrosis by regulating the ACE1/AngⅡ/AT1R axis， increasing the relative abundance of Lactobacillus and Bifidobacterium， reducing the relative abundance of Moralella， improving the richness and evenness of intestinal flora， and alleviating pathological damage in the renal tissue.

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 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.

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.

Inhibition of procyanidin from Vaccinium vitisidaea on proliferation of cardiac fibroblast in neonatal rats and its mechanism / 中草药

Objective To investigate the effect of procyanidin from Vaccinium vitisidaea on the proli-feration of cardiac fibroblast(CFb) induced by angiotensin Ⅱ(AngⅡ),and to explore its mechanism.Methods The CFb proliferation of cultured neonatal rat was induced by AngⅡ and detected by MTT assay.The levels of collagen Ⅰ,collagen Ⅲ,and TGF-?1 were measured by ELISA.The change of NO content and iNOS activity were measured by nitric acid reductase method and spectrophotometry.Cell cycle was assessed via flow cytometry(FCM). The expression of cell cycle regulatory protein p27 was determined by the combination of immunocytochemical staining and image analysis software.Results CFb Proliferation,collagen content,and TGF-?1 levels in culture medium were markedly inhibited when CFb were treated with procyanidin at concentration of 25,50,and 100 mg/L(P

Effect of pronuciferine on apoptosis of human umbilical vein endothelium cells induced by angiotensin Ⅱ / 中草药

Objective To investigate the effect of pronuciferine on apoptosis of cultured human umbilical vein endothelium cells(HUVECs) induced by angiotensin Ⅱ(AngⅡ).Methods HUVECs cell line ECV304 was cultured in vitro,pretreated with Captopril(10 ?mol/L) or pronuciferine 10,1,0.1,0.01 ?mol/L for 30 min,respectively,then treated with AngⅡ(1 ?mol/L).Cell-morphosis was observed by light microscope.Cells viability was assessed by MTT assay.Production of nitric oxide(NO),activities of total nitric oxide synthase(tNOS),and inducible nitric oxide synthase(iNOS) were measured by colorimetry.Apoptosis rate was measured by Flow Cytometer(FCM).Results AngⅡ induced typical endothelial cell apoptosis and the apoptosis rates were significantly higher than those of the control group((P

Inhibitory effects of chitosan on the proliferation of bovine basilar artery vascular smooth muscle cells induced by angiotensin Ⅱ / 中国海洋药物

The bovine basilar artery vascular smooth muscle cells(BAVSMC S) were isolated and cultured.The effects of chitosan and enalaprilat on the proliferation of BAVSMC S induced by angiotensin Ⅱ(AngⅡ) were evaluated with MTT method,the DNA synthesis in BAVSMC S was measured by thymidine incorporation and total protein of BAVSMC S was detected by coomassie brilliant blue method.The results showed that chitosan can inhibit the proliferation of BAVSMC S induced by AngⅡ in a time and concentration dependent manner,and depress thymidine incorporation of BAVSMC S induced by AngⅡ in a concentration dependent manner chitosan can reduce the content of protein in BAVSMC S induced by AngⅡ.These findings suggested that chitosan is capable of producing an inhibitory effect on proliferation of bovine basilar artery vascular smooth muscle cells induced by AngⅡ .

Studies on mechanism of antihypertension effect of extracts from Compound Jueming / 中草药

Object To probe into the antihypertension activity mechanism of extracts from Compound Jueming (CJ). Methods The method of radioimmunoassay was used to assay the concentration of the related substances on spontaneously hypertensive rats (SHR) such as angiotensin Ⅱ (ANGⅡ), renin activity (RA), endothelin (ET), atrial natriuretic polypeptide (ANP), aldosterone (ALD), and urine. Results All of the three doses (9.5, 7.2, and 4.8 g/kg) of extracts from CJ reduced ANG Ⅱ and RA, but had no influence on ALD, ET, ANP, and urine in SHR. Conclusion The antihypertension effect of extracts from CJ is related to decreasing of renin angiotensin, but not to ALD, ET, ANP, and urine.

Effects of taurine on left ventricular hypertrophy in renovascular hypertensive rats / 中国药理学通报

AIM To investigate the preventive effects of taurine on the left ventricle hypertrophy in renovascular hypertensive rats. METHODS Two-kidney 1-clip (2K1C) rats were used to establish the model of renovascular hypertension. The myocardial local aldosterone(Ald) and AngⅡconcentration (MDC,MAC) were measured by radioimmunoassay. Expression of oncogene c-fos mRNA was analyzed by means of in situ hybridization. The myocardial collagen concentration (MCC) was measured by biochemical method. The myocardial tissue structure was observed under the microscope ,and the myocardial fiber diamension (MFD) was measured with micrometer. The linear correlation between MCC, MFD and MAC or MDC was analysed respectively. RESULTS Compared with sham operated rats ,the MCC, MAC, MDC and MFD were significantly increased in 2K1C rats ( P