Changes in Intestinal Microflora and Lipid Metabolism in Ovariectomized Rats and Regulatory Effect of Erxian Decoction / 中国实验方剂学杂志

ObjectiveTo explore the intervention effect of Erxian decoction on intestinal microflora after ovariectomy in rats by 16S rRNA gene sequencing. MethodThirty-two female healthy SD rats were randomly divided into a Sham operation （Sham） group， a model （OVX） group， an estrogen （E） group， and an Erxian decoction （EXD） group， with 8 rats in each group. The rats in the E group and the EXD group received 1.8×10-4 g·kg-1 estradiol valerate solution and 9 g·kg-1 Erxian decoction， respectively， and those in the Sham group and the OVX group received an equal volume of distilled water once a day for 16 weeks. After 16 weeks， the levels of serum estrogen and blood lipid were detected. The fecal DNA was extracted， followed by 16S rRNA gene sequencing and analysis. ResultCompared with the Sham group， the OVX group showed reduced serum estrogen level （P<0.01） and increased serum levels of total cholesterol （TC） and low-density lipoprotein cholesterol （LDL-C） （P<0.05）. Compared with the OVX group， the E group and the EXD group showed increased serum estrogen level （P<0.01） and reduced TC and LDL-C （P<0.05）. Alpha diversity showed that there was no significant change in intestinal microflora diversity after ovariectomy. Beta diversity showed that there were significant differences in the structure of intestinal microflora in the four groups. The intervention of Erxian decoction could improve the changes in intestinal microflora after ovariectomy. LEfSe was used to analyze the differential flora in the four groups. The results showed that the Sham group and the OVX group had 3 differential bacterial phyla and 18 differential bacterial genera， the OVX group and the E group had 1 differential bacterial phylum and 12 differential bacterial genera， and the OVX group and the EXD group had 3 differential bacterial phyla and 5 differential bacterial genera. Estrogen intervention could reverse the change trend of Ruminococcus 1， Anaerovibrio， and Turicibacter in the OVX group. Erxian decoction intervention could reverse the change trend of Bacteroidetes， Firmicutes， Prevotella 9， Ruminococcaceae UCG-014， Ruminococcus 1， and Fusicatenibacter in the OVX group. ConclusionThe structure and function of intestinal microflora in ovariectomized rats changed obviously， and Erxian decoction could ameliorate the change.

Effect of Erxiantang on Cardiac Function and Blood Pressure Dynamic Evolution in Ovariectomized Rats / 中国实验方剂学杂志

Objective::To investigate the evolution of cardiac function and blood pressure in ovariectomized rats and the effect and mechanism of Erxiantang. Method::Healthy 10-week-old female SPF SD rats were randomly divided into sham operation group, model group, estrogen group(estradiol valerate, 0.18 mg·kg-1) and Erxiantang group(7.5 g·kg-1). The rats were intragastrically administered 2 weeks after ovariectomy, once a day for 12 weeks.Sham operation groups and model groups were given equal volumes of purified water.At the 4th week, 8th week, and 12th week after administration, the cardiac function, blood pressure, and levels of estrogen (E2) in rat serum were measured by non-invasive ultrasound cardiogram (UCG), tail artery detection techniques and radioimmunoassay.The levels of endothelin-1 (ET-1) and angiotensin 2(Ang Ⅱ) in rat serum were detected by enzyme-linked immuno sorbent assay (ELISA). The cardiac morphology and apoptosis were detected by hematoxylin-eosin (HE) staining, electron microscopy and Terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL). Result::Compared with sham operation group, the ejection fraction (EF) decreased and the left ventricular end systolic volume (LVVols) increased in the model group at 4th week after administration(P<0.05). There was no significant difference in cardiac function between the groups at 8th week.The left ventricular end diastolic diameter (LVIDs), LVVols, left ventricular end diastolic diameter (LVIDd), and left ventricular end diastolic volume (LVVold) were significantly increased in the model group at 12th week (P<0.01). At the 4th weeks, 8th week and 12th week, the systolic blood pressure (SBP) of the model group increased (P<0.05) and showed an increasing trend, and the diastolic blood pressure (DBP) did not change significantly.At the 12th week, the levels of E2 in serum decreased (P<0.05), ET-1 and Ang Ⅱ increased of the model group (P<0.01). The cardiac myofibrils were irregular, some myofilament was broken, and mitochondrial palsy was disordered, broken or disappeared, and cardiac apoptosis increased (P<0.01). Compared with the model group, myocardial contraction and diastolic function were significantly improved in Erxian decoction group, and blood pressure was decreased.The levels of E2 in serum was increased (P<0.05). The levels of ET-1 was decreased (P<0.05), and AngⅡ in serum was significantly decreased (P<0.01). The mitochondrial morphological structure was improved and the cardiac apoptotic rate was significantly decreased (P<0.01). Conclusion::After the ovariectomy, the rats showed a series of pathological changes such as decreased heart function and increased blood pressure.Compared with the decrease of heart function, the changes of blood pressure appeared earlier.Erxiantang exerts its intervention on cardiac function and blood pressure in ovariectomized rats by regulating E2, blood active substances and cardiac apoptosis.

Effect of Erxian Decoction on Myocardial Microvessels and Hemorheology in Ovariectomized Rats / 中国实验方剂学杂志

Objective:To observe the changes of myocardial microvessel density， microvascular endothelial cell morphology and hemorheology in ovariectomized rats and explore the interventional effects of Erxian decoction. Method:Thirty-two healthy 10 week-old female SPF SD rats were randomly divided into sham operation group， model group， estrogen group （estradiol valerate， 0.18 mg·kg-1） and Erxian decoction group （9 g·kg-1）. The rats were intragastrically administered 2 weeks after ovariectomy， once a day for 16 weeks. Sham operation groups and model groups were given equal volumes of purified water. After 16 weeks of administration， the cardiac function was measured by noninvasive ultrasound cardiogram （UCG）， CD34 in the myocardial tissue was tested by immunofluorescence staining to measure the microvessel density， the morphological structure of microvessels of myocardial tissue were detected by transmission electron microscope， the levels of estrogen （E2） in rat plasma were detected by radioimmunoassay， the levels of endothelin-1 （ET-1）， prostacyclin I2 （PGI2）， thromboxane A2 （TXA2）， endothelial growth factor （VEGF）， and von Willebrand Factor （vWF） in rat plasma were detected by enzyme-linked immuno sorbent assay （ELISA）， four items of coagulation was detected by blood coagulation analyzer， whole blood viscosity and plasma viscosity were detected by hemorheology. Result:Compared with sham operation group， the ejection fraction （EF） decreased （P<0.01）， the left ventricular short axis shortening rate （FS） decreased （P<0.01）， and the left ventricular end systolic volume （LVVols） increased （P<0.01）， myocardial microvessel density significantly reduced （P<0.01）， the endothelial cells were swollen and the cytoplasm was cavitation， E2 in rat plasma decreased （P<0.01）， ET-1， VEGF， vWF increased （P<0.01）， prostacyclin I2 /thromboxane A2 （PGI2/TXA2） decreased （P<0.01）， plasma activated partial prothrombin time （APTT） decreased （P<0.01）， fibrinogen （FIB） increased （P<0.01）， whole blood viscosity， plasma viscosity， and cassone viscosity increased （P<0.01）， whole blood high-cut， low-cut index， and red blood cell （RBC） aggregation index increased （P<0.05） in model group. Compared with model group， EF and FS increased （P<0.05）， LVVols decreased （P<0.05）， myocardial microvessel density significantly increased （P<0.01）， the endothelial cell edema was improved， and transport vesicles were clearly visible， E2 in rat plasma increased （P<0.01）， ET-1， VEGF， decreased （P<0.01）， PGI2/TXA2 increased （P<0.01）， APTT increased （P<0.01）， whole blood viscosity， whole blood high shear relative index， RBC aggregation index decreased （P<0.05）， Kasson viscosity and plasma viscosity decreased （P<0.01） in Erxian decoction group. Conclusion:Erxian decoction increases myocardial microvessel density， protects the structural integrity of microvascular endothelial cells， improves its endothelial secretion function and hemorheology in ovariectomized rats， and protects heart function.

Expression and roles of long non-coding RNA ZFAS1 in non-small cell lung cancer / 临床检验杂志

Objective To determine the expression levels of zinc finger antisense 1 (ZFAS1) in tumor tissues of non-small cell lung cancer (NSCLC) and investigate the biological roles of ZFAS1 in NSCLC.Methods The relative expression levels of ZFAS1 in tumor tissues of NSCLC patients were determined by using real-time fluorescent qRT-PCR.RNA interference was used to knock down ZFAS1 expression in A549 cells.The proliferations of A549 cells with ZFAS1 knockdown were determined by cell counting and cell colony formation assays.Flow cytometric analysis was used to determine the cellular cycle distribution and apoptosis of the A549 cells with ZFAS1 knockdown.Transwell migration and matrigel invasion assays were used to determine the migration and invasion of the A549 cells with ZFAS1 knockdown.The gene expression levels of cyclin D1,Bcl2,N-cadherin,ZEB1,Slug and Twist were determined by real-time fluorescent qRT-PCR.Results The mean expression levels of ZFAS1 in tumor tissues of NSCLC patients [0.01 (0.002 to 0.054)] were significantly higher than those in the adjacent non-cancerous tissues [0.002 (0.001 to 0.012)] (Z =-2.638;P ＜ 0.01).After ZFAS1 knockdown,the proliferation of A549 cells was remarkably retarded (P ＜ 0.01).The percentage of cells at G1 phase was increased while the cells at S phase was decreased (P ＜ 0.01).The rate of apoptotic cells was significantly increased in A549 cells with ZFAS1 knockdown (P ＜ 0.01).A549 cells showed decreased migration and invasion abilities after ZFAS1 knockdown (P ＜0.01).The expression levels of cyclin D1,Bcl2,N-cadherin,ZEB1,Slug and Twist genes were decreased in ZFAS1 knockdown A549 cells (P ＜ 0.05).Conclusion ZFASI was highly expressed in the tumor tissues of NSCLC patients.ZFAS1 knockdown induced cell cycle arrest,cell apoptosis and suppression of epithelial-mesenchymal transition (EMT),leading to the inhibition of proliferation,migration,and invasion of NSCLC cells.

Effects of Shenmai injection on afterdepolarization and triggered activities in left ventricular papillary muscle in rat cardiac hypertrophy / 中国中药杂志

This study is to evaluate the effects of Shenmai injection on the temporal alterations of action potential (AP), early afterdepolarization (EAD) and delayed afterdepolarization (DAD) in papillary muscles. The action potentials were recorded by a glass electrode. APD at 90% repolarization (APD9 ) was measured, and spontaneous EAD and DAD were observed. The results show APD90 was significantly prolonged in model group compared with sham-operated group, whereas it was remained unchanged in Shenmai injec- tion treatment group and amiodarone group. The spontaneous EADs and DADs were frequently visible in model group. In conclusion, EAD, DAD and trigger activities increase gradually during pathological progression of rat cardiac hypertrophy, and Shenmai injection could improve the action potential change in rat cardiac hypertrophy.

Electrophysiological effects of antiarrhythmic drugs on pacemaker cells in guinea pig left ventricular outflow tract / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the electrophysiological effects of antiarrhythmic drugs on pacemaker cells of left ventricular outflow tract.</p><p><b>METHODS</b>By using conventional intracellular microelectrode technique to record action potentials, series antiarrhythmic drugs were used to investigate the electrophysiological features and regularities of spontaneous activity of left ventricular outflow tract.</p><p><b>RESULTS</b>(1) Perfusion with 1 micromol/L quinidine resulted in a significant decrease in rate of pacemaker firing (RPF, P < 0.05), velocity of diastolic depolarization (VDD, P < 0.05), amplitude of action potential (APA, P < 0.05), and maximal rate of depolarization (V(max), P < 0.05), and a marked prolonging in 50% and 90% of duration of action potential (APD50 and APD90, P < 0.05). (2) 1 micromol/L lidocaine decreased RPF, VDD, MDP, APA and V(max) significantly (P < 0.05), shortened APD50 and APD90 notably (P < 0.05). (3) 1 micromol/L propafenone led to a significant decrease in RPF (P < 0.01), VDD (P < 0.05), APA (P < 0.05), V(max) (P < 0.01), and a marked prolonging in APD50 (P < 0.01) and APD90 (P < 0.05). (4) Application of 5 micromol/L propranolol resulted in a significant decrease in RPF and VDD (P < 0.01), MDP and APA (P < 0.01), V(max) (P < 0.05) and a notable prolonging in APD50 and APD90 (P < 0.05). (5) Perfusion with 1 micromol/L amiodarone resulted in a significant decrease in RPF and VDD (P < 0.01), APA (P < 0.01), V(max) (P < 0.05), a marked prolonging in APD50 (P < 0.01) and APD90 (P < 0.05). (6) 1 micromol/L verapamil significantly decreased RPF and VDD (P < 0.01), MDP and APA (P < 0.05), V(max) (P < 0.05), notably prolonged APD50 and APD90 (P < 0.01). (7) 50 micromol/L adenosine significantly decreased RPF and VDD (P < 0.05), APA (P < 0.05), V(max) (P < 0.01), markedly shortened APD50 and APD90 (P < 0.05).</p><p><b>CONCLUSION</b>All kinds of antiarrhythmic drugs can decrease the autorhythmicity of guinea pig left ventricular outflow tract. By altering APD50 and APD90, they can affect effective refractory period (ERP) and having a significant effect on autorhythmicity of left ventricular outflow tract.</p>

Effects of Panax notoginseng saponins on rat cardiomyocytes apoptosis induced by angiotengin II in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To study the protective effects of Panax notoginseng saponins (PNS) on angiotensin II (Ang II)-induced rat cardiomyocyte apoptosis in vitro and the probable mechanism.</p><p><b>METHOD</b>Cultured cardiomyocytes from neonatal rats were stimulated with Ang II. Cell viability was measured by MTT. Apoptosis was evaluated using Acridine Orange (AO) fluorescent dye staining and flow cytometry; Fluo-3 AM was used to test the change of intracellular free calcium.</p><p><b>RESULT</b>It was found that incubating with Ang II (10(-7) mol x L(-1)) for 48 h increased cardiomyocyte apoptosis, PNS (25, 100 mg x mL(-1)) increased myocyte viability. PNS (50 mg x mL(-1)) significantly decreased this Ang II-induced rat cardiomyocyte apoptosis (P < 0.05) and decreased fluorescent intensity of intracellular calcium.</p><p><b>CONCLUSION</b>PNS has a significant effect on Ang II-induced rat cardiomyocytes apoptosis in vitro by alleviating intracellular calcium overload.</p>

Electrophysiological effects of neurotransmitters on pacemaker cells in guinea pig left ventricular outflow tract / 生理学报

This study was designed to explore the innervation of autonomic nervous system and the distribution of receptors on pacemaker cell membrane in guinea pig left ventricular outflow tract (aortic vestibule). By using conventional intracellular microelectrode technique to record action potentials, autonomic neurotransmitters and antagonists were used to investigate the electrophysiological features and regularities of spontaneous activity of left ventricular outflow tract cells. Electrophysiological parameters examined were: maximal diastolic potential (MDP), amplitude of action potential (APA), maximal rate of depolarization (V(max)), velocity of diastolic depolarization (VDD), rate of pacemaker firing (RPF), 50% and 90% of duration of action potential (APD(50) and APD(90)). The results are listed below: (1) Perfusion with 100 mumol/L isoprenaline (Iso) resulted in a significant increase in V(max) (P <0.05), VDD, RPF, and APA (P <0.01), a notable decrease in MDP (P<0.05), and also a marked shortening in APD(50) (P<0.01). Pretreatment with Iso (100 mumol/L), propranolol (5 mumol/L) significantly decreased RPF and VDD (P<0.01), decreased APA, MDP and V(max) (P<0.01) notably, prolonged APD(50) (P<0.01) and APD(90) (P<0.05) markedly. (2) Application of 100 mumol/L epinephrine (E) resulted in a significant increase in VDD (P<0.05), RPF (P<0.001), V(max) (P<0.05) and APA (P<0.001), and a notable shortening in APD(50) and APD(90) (P<0.05). (3) Perfusion with 100 mumol/L norepinephrine (NE) led to a significant increase in VDD, RPF, APA and V(max) (P<0.05), and a marked shortening in APD(50) (P<0.05). Pretreatment with NE (100 mumol/L), phentolamine (100 mumol/L) significantly decreased RPF and VDD, MDP and APA (P<0.01), decreased V(max) notably (P<0.05), prolonged APD(50) and APD(90) markedly (P<0.01). (4) During perfusion with 10 mmol/L acetylcholine (ACh), VDD and RPF slowed down notably (P<0.05), APA decreased significantly (P<0.001), V(max) slowed down notably (P<0.01), APD50 shortened markedly (P<0.05), Atropine (10 mmol/L) antagonized the effects of ACh (10 mumol/L) on APD(50) (P<0.05). These results suggest that there are probably alpha-adrenergic receptor (alpha-AR), beta-adrenergic receptor (beta-AR) and muscarinic receptor (MR) on pacemaker cell membrane of left ventricular outflow tract in guinea pig. The spontaneous activities of left ventricular outflow tract cells are likely regulated by sympathetic and parasympathetic nerves.

Comparison of the electrophysiological features between the rhythmic cells of the aortic vestibule and the sinoatrial node in the rabbit / 生理学报

The purpose of this study was to clarify the characteristics of the pacemaker cells in the left ventricular outflow tract (aortic vestibule) and compare them with those of the cells in the sinoatrial node (SAN). By using conventional intracellular microelectrode technique to record their action potentials, some ionic channel blockers were used to observe their electrophysiological effects on the two types of pacemaker cells in the rabbit, especially on the ionic movement during phase 0 and phase 4. The results obtained are as follows. (1) Perfusion with 1 micromol/L verapamil (VER) resulted in a significant reduction in the amplitude of action potential (APA), maximal rate of depolarization (V(max)), absolute value of the maximal diastolic potential (MDP), velocity of diastolic depolarization (VDD) and rate of pacemaker firing (RPF), and also a prolongation of the 90% of the duration of action potential (APD(90)) in the pacemaker cells of the SAN and aortic vestibule (P<0.05). (2) Perfusion with 180 micromol/L nickel chloride (NiCl2) resulted in a decrease in VDD in the two types of the pacemaker cells (P<0.01). APA, V(max) and RPF fell notably, and the APD(90) prolonged in the sinoatrial node cells (P<0.05). (3) 2 mmol/L 4-aminopyridine (4-AP) led to a increase in VDD in both types of pacemaker cells (P<0.01). At the same time the absolute values of MDP, APA and V(max) decreased significantly, and APD(90) prolonged notably (P<0.05). During the perfusion, RPF in SAN increased markedly, while RPF in aortic vestibule exhibited no significant change. (4) 2 mmol/L cesium chloride (CsCl) led to a decrease in VDD and RPF in the two types of the pacemaker cells (P<0.05).These results suggested: (1) the ion currents in phase 0 and phase 4 of depolarization and repolarization of slow-response activity in aortic vestibule are similar to those in dominant pacemaker cells of sinoatrial node; (2) for the pacemaker cells in the left ventricular outflow tract, Ca(2+) current is the main depolarizing ion current of the phase 0, K(+) current is the main factor responsible for the repolarization. Attenuation of K(+) current is responsible for the phase 4 spontaneous depolarization. In addition, it seems that I(Ca-T), I(Ca-L) and I(f ) play some role in the pacemaker currents.