Astragaloside IV alleviates heart failure via activating PPARα to switch glycolysis to fatty acid ß-oxidation.

In heart failure (HF), energy metabolism pathway in cardiac muscle changes from fatty acid ß-oxidation to glycolysis. However, the exact mechanism is unknown. Sarcoendoplasmic reticulum Ca2+α ATPase (SERCA) expression is downregulated and mitochondrial function is reduced in HF, perhaps partly due to a substantially reduced energy supply for excitation-contraction coupling resulting from a lower fatty acid ß-oxidation rate. We investigated whether Astragaloside IV can activate peroxisome proliferator-activated receptor alpha (PPARα) to stimulate fatty acid ß-oxidation and increase cardiac energy production, improving mitochondrial function and the efficiency of SERCA in HF. In pressure overload-induced HF mice and isolated hypertrophic myocardial cells, fatty acid ß-oxidation and heart function were substantially strengthened following Astragaloside IV treatment, as demonstrated by the increased expression of PPARα and SERCA2a. In vitro, Astragaloside IV regulated energy metabolism by increasing ATP production and enhancing mitochondrial function, attributable to increased oxygen consumption and slightly increased mitochondrial Ca2+ uptake. In HF, Astragaloside IV switched glycolysis to fatty acid ß-oxidation, as confirmed by reduced anaerobic glycolysis and an increased oxygen consumption ratio. These results suggest that Astragaloside IV can stimulate fatty acid ß-oxidation and improve mitochondrial function, which may present a novel cardioprotective treatment that inhibits the progress of HF.

Stachydrine Protects Against Pressure Overload-Induced Cardiac Hypertrophy by Suppressing Autophagy.

BACKGROUND: Autophagy is required for the maintenance of cardiomyocyte homeostasis. However, excessive autophagy plays a maladaptive role in pressure overload-induced heart failure. To identify mechanisms by which Stachydrine inhibits pressure overload-induced cardiac hypertrophy, we determined inhibitory activities against activation of NADPH oxidase, reactive oxygen species(ROS) production and excessive activation of autophagy. METHODS: Stachydrine was administered intragastrically to Wistar rats after Transverse aortic constriction(TAC) and H9c2 cells were treated with Stachydrine after Angiotension II stimulation. The activation of NADPH oxidase2 required the membrane translocation of p47phox and p67phox. Cell membrane fraction was isolated by ultracentrifuge in sucrose. The expression of p67phox, p47phox, gp91phox subunit in the cell membrane were determined by western blot. The combination of p67phox and gp91 phox subunit was detected by immunofluorescence staining. The expression of phosphorylated p47phox subunit was determined by western blot. The intracellular ROS were measured with DCF-DA fluoresence. The autophagic flux was measured by recording the fluorescence emission of the fusion protein mRFP-GFP-LC3 by dynamic live-cell imaging. Reuslts: We report here that stachydrine, a major constituent of Leonurus heterophyllus Sweet, inhibited AngII-induced excessive autophagy within H9c2 cells. Stachydrine blocked the over phosphorylation of the p47phox subunit, decreased the translocation of p47phox and p67phox to the membrane, inhibited the activity of NOX2, and reduced the generation of ROS. We also demonstrated that stachydrine ameliorated TAC-induced cardiac hypertrophy, dysfunction and excessive autophagy in vivo. CONCLUSIONS: Our study highlights the importance of regulating NOX2 when autophagy is obviously activated. By inhibiting NOX2, Stachydrine inhibits ROS production, thus exerting a remarkable activity of inhibiting hypertrophy, which could have considerable effect on clinical practice.

The Zuo Jin Wan Formula Induces Mitochondrial Apoptosis of Cisplatin-Resistant Gastric Cancer Cells via Cofilin-1.

Despite the status of cisplatin (DDP) as a classical chemotherapeutic agent in the treatment of cancer, the development of multidrug resistance often leads to a failure of DDP therapy. Here we found that phosphorylated cofilin-1 (p-cofilin-1) was overexpressed in the DDP-resistant human gastric cancer cell lines SGC7901/DDP and BGC823/DDP, relative to the respective parent cell lines (SGC7901 and BGC823), and that DDP induced the dephosphorylation of p-cofilin-1 in both parent lines but not in the DDP-resistant lines. However, we noted that the traditional Chinese medicine formula Zuo Jin Wan (ZJW) could induce the dephosphorylation of p-cofilin-1 and promote cofilin-1 translocation from the cytoplasm into the mitochondria in both SGC7901/DDP and BGC823/DDP cells. This mitochondrial translocation of cofilin-1 was found to induce the conversion of filamentous actin to globular-actin, activate mitochondrial damage and calcium overloading, and induce the mitochondrial apoptosis pathway. We further observed that these effects of ZJW on DDP-resistant human gastric cancer cell lines could be reversed via transfection with cofilin-1-specific siRNA, or treatment with a PP1 and PP2A inhibitor. These results suggest that ZJW is an effective drug therapy for patients with DDP-resistant gastric cancer.

Shengmai San Ameliorates Myocardial Dysfunction and Fibrosis in Diabetic db/db Mice.

In this study, we mainly investigated the effects of Shengmai San (SMS) on diabetic cardiomyopathy (DCM) in db/db mice. The db/db mice were randomly divided into model group and SMS group, while C57BLKS/J inbred mice were used as controls. After 24-week treatment, blood glucose, body weight, and heart weight were determined. Hemodynamic changes in the left ventricle were measured using catheterization. The myocardial structure and subcellular structural changes were observed by HE staining and electron microscopy; the myocardium collagen content was quantified by Masson staining. To further explore the protective mechanism of SMS, we analyzed the expression profiles of fibrotic related proteins. Compared to nondiabetic mice, db/db mice exhibited enhanced diastolic myocardial dysfunction and adverse structural remodeling. Higher expression of profibrotic proteins and lower levels of extracellular matrix degradation were also observed. After SMS oral administration for 24 weeks, cardiac dysfunction, hypertrophy, and fibrosis in diabetic mice were greatly improved. Moreover, increased profibrotic protein expression was strongly reversed by SMS treatment in db/db mice. The results demonstrate that SMS exerts a cardioprotective effect against DCM by attenuating myocardial hypertrophy and fibrosis via a TGF-ß dependent pathway.

Effects of stachydrine on norepinephrine-induced neonatal rat cardiac myocytes hypertrophy and intracellular calcium transients.

BACKGROUND: Leonurus heterophyllus sweet has been suggested to have cardioprotective effects against heart diseases, including ischemic diseases and ventricular remodeling. However, the active ingredients of the herb and the underlying mechanisms are poorly understood. The aim of the present study was to investigate the effects of stachydrine (STA), a major constituent of Leonurus heterophyllus sweet, on norepinephrine (NE) induced hypertrophy and the changes of calcium transients in neonatal rat cardiomyocytes. METHODS: Ventricular myocytes from 1-day-old Wistar rats were isolated and cultured in DMEM/F12 with 1 µmol/L norepinephrine in the presence or absence of 10 µmol/L STA for 72 h. Cardiomyocytes hypertrophy was evaluated by cell surface area, total protein/DNA content, ß/α-MHC mRNA ratio. While calcium handling function was evaluated by Ca2+-transient amplitude and decay, SERCA2a activity and expression, PLN expression and phosphorylation. ß1-adrenergic receptor system activation was evaluated by the content of cAMP and the activation of PKA. RESULTS: NE treatment increases the cell surface area, protein synthesis, the expression level of ß-MHC and ß/α-MHC ratio. These effects were attenuated by STA. NE-induced hypertrophy was associated with increased Ca2+-transient amplitude, accelerated decay of the Ca2+-transient, increased phospholamban expression, hyper-phosphorylation at both the serine-16 and threonine-17 residues, increased intracellular cAMP level, and PKA overactivation. All of which were significantly inhibited by STA. CONCLUSION: These data suggest that STA attenuates norepinephrine-induced cardiomyocyte hypertrophy and has potential protective effects against ß-adrenergic receptor induced Ca2+ mishandling.

[Effects of buyang huanwu decoction on the sarcoplasmic reticulum calcium uptake in abdominal aortic constriction induced myocardial hypertrophic rats].

UNLABELLED: objective: To investigate effects of buyang huanwu decoction (BYHWD) on the rats' myocardial hypertrophic model induced by abdominal aortic constriction, and to clarify the molecular regulatory mechanisms for sarcoplasmic reticulum calcium uptake. METHODS: Hypertrophic myocardium rat model was induced by abdominal aorta constriction (AAC). Four weeks after modeling, rats were randomly divided into the sham-operation group (Group S), the AAC model group (Group M), the Enalapril group (Group E), and the BYHWD treatment group (Group BYHWD), respectively. The left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), + dp/dtmax,-dp/dtmax, cardiac output (CO), heart mass index (HMI), and left ventricular mass index (LVMI) were observed in each group after 12-week medication. The serum contents of the atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) were detected using ELISA. The SERCA2a activity, the ex- pressions of SERCA2a, phospholamban (PLN), and PLN phosphorylation were observed finally. RESULTS: Compared with Group S, LVSP and LVEDP significantly increased,-dp/dtmax and CO obviously decreased, the myocardial tissue was obviously thickened, the serum contents of ANP and BNP increased, the activity and expression of SERCA2a decreased, the SERCA2a/PLN ratio and PLN phosphorylation degree decreased in Group M (all P <0.05). Compared with Group M, LVEDP obviously decreased, -dp/dtmax and CO obviously increased, the hypertrophy myocardial tissue was obviously lessened, the serum contents of ANP and BNP decreased, the activity of SERCA2a increased, the relative expression contents of SERCA2a, Ser16, and Thrl7 were elevated in Group BYHWD (all P <0.05). BYHWD had significant roles in elevating the SERCA2a/PLN ratio and PLN phosphorylation degree (P <0.05). CONCLUSION: BYHWD could significantly improve hemodynamics of heart failure rats, elevate CO, lessen cardiac hypertrophy, and improve the capabilities for sarcoplasmic reticulum calcium uptake.

[Effect of Leonurus stachydrine on myocardial cell hypertrophy].

OBJECTIVE: To investigate the effects of Leonurus stachydrine on myocardial cell hypertrophy induced by angiotensin II in rats. METHODS: Myocardial cells of neonatal rats were incubated with angiotensin II for 48 hours to establish myocardial cell hypertrophy models. The cell surface area and protein/DNA ratio were detected as index for hypertrophy of myocardial cells. ROS positive myocardial cell were counted by means of flow cytometry, the expression of p-IkappaB (ser32) protein in the cytosol and NF-kappaB (p65) protein in the nucleus were also detected by Western blot. RESULTS: Various concentrations of stachydrine intervention (10(-6) - 10(-4) mol/L) showed better anti-hypertrophic effect and decreased the number of ROS positive myocardial cell (P<0.05). Meanwhile, intervention of stachydrine significantly suppressed the level of p-IkappaB (ser32) protein in the cytosol and NF-kappaB (p65) protein in the nucleus (P<0.05). CONCLUSION: Stachydrine can inhibit the NF-kappaB signal pathway, and this may be ralated to the mechanism of anti-hypertrophic.

[Research progress of sarcolipin-a new regulatory protein of sarcoplasmic reticulum Ca2+ ATPase].

Sarcolipin (SLN) is a 3 kD membrane protein found in sarcoplasmic reticulum (SR). It has 31 amino acid residues; SLN and phopholamban (PLB) are belong to the same protein family, so they have similar physiological functions. SLN inhibits sarcoplasmic reticulum Ca(2+) ATPase (SERCA) activity and reduces its affinity of Ca(2+), resulting in dysfunction of myocardial contraction and heart failure. However, much remains to be elucidated. SLN independently or in conjunction with PLB affects SERCA activity, imbalancing intracellular calcium homeostasis, and reducing myocardial contractivity; these effects promote the development of heart failure.

[Protective effects of Leonurus japonicas on myocardial remodeling induced by isoproterenol in rats].

OBJECTIVE: To investigate the effects of Leonurus japonicas on myocardial remodeling induced by isoproterenol (ISO) in rats. METHODS: The model of rat myocardial remodeling was established by subcutaneous injection of ISO (20,10 and 5 mg/kg for 3 days, subsequently 3 mg/kg for 7 days). Rats were randomly divided into five groups: Control, ISO, ISO + enalapril, ISO + Leonurus 8 g/ kg and ISO + Leonurus 16 g/kg. Recorded the values of LVSP, LVEDP, +/- dp/dt (max), cardiac output ( CO). Weighed the body weight (BW), heart weight (HW) and left ventricular weight (LVW) of rats to calculate the values of HW/BW and LVW/BW. The hydroxyproline contents were investigated by spectrophotometric measurement. The contents of type I collagen and type III collagen and ratio of I/III collagen were assessed by immunohistochemical stain. RESULTS: Leonurus (16 g/kg x day(-1)) could increased the value of LVSP, + dp/dt(max), CO (P < 0.05). Leonurus (8 g/kg) could increased the value of -dp/dt(max), decreased the ratios of HW/BW and LVW/BW (P < 0.05), the contents of hydroxyproline, type I collagen, type III collagen and the ratios of I/III collagen (P < 0.05). CONCLUSIONS: Leonurus at a dosage of 16 g/kg may improve the systolic function; Leonurus at a dosage of 8 g/kg may improve the diastolic function, down-regulate the expression of collagen and normalize the ratio of I/III collagen.

[Dynamic study on inhibition of qi-benefiting, blood-activating recipe on myocardial hypertrophy induced by ISO in rats].

OBJECTIVE: To observe the effect of Qi-Benefiting, Blood-Activating Recipe on the different pathologic stage of myocardial hypertrophy induced by ISO in rats. METHODS: Myocardial hypertrophy rats were induced by isoproterenol, and treated with Qi-Benefiting, Blood-Activating Recipe for 5,10 and 15 weeks, hemodynamic parameters, LVMI, HMI were determined, ANP and BNP were analysed. RESULTS: Qi-Benefiting, Blood-Activating Recipe could increase cardiac output and improve hemodynamic parameters all after 5, 10 and 15 weeks' treatment (P<0.05). It could decrease the contents of ANP and BNP after 5, 10 and 15 weeks' treatment (P<0.05). Qi-Benefiting, Blood-Activating Recipe could significantly decrease the levels of HWI and LVMI after 5, 10 and 15 weeks' treatment (P<0.05). CONCLUSION: Qi-Benefiting, Blood-Activating Recipe can significant improve hemodynamic status, increase cardiac output and decrease the level of neurohormonal factors.

Lipopolysaccharide "two-hit" induced refractory hypoxemia acute respiratory distress model in rats.

To establish a stable and reliable model of refractory hypoxemia acute respiratory distress syndrome (ARDS) and examine its pathological mechanisms, a total of 144 healthy male Wistar rats were randomized into 4 groups: group I (saline control group), group II (LPS intravenous "single-hit" group), group III (LPS intratracheal "single-hit" group) and Group IV (LPS "two-hit" group). Rats were intravenously injected or intratracheally instilled with a large dose of LPS (10 mg/kg in 0.5 mL) to simulate a single attack of ARDS, or intraperitoneally injected with a small dose of LPS (1 mg/kg) followed by tracheal instillation with median dose of LPS (5 mg/kg) to establish a "two-hit" model. Rats in each group were monitored by arterial blood gas analysis and visual inspection for three consecutive days. Arterial blood gas values, lung wet/dry weight ratio and pathological pulmonary changes were analyzed to determine the effects of each ALI/ARDS model. Concentrations of TNF-alpha, IL-1 and IL-10 in the bronchoalveolar lavage fluid (BALF) and blood plasma were measured by using enzyme-linked immunosorbent assays (ELISA). Our results showed that single LPS-stimulation, whether through intravenous injection or tracheal instillation, could only induce ALI and temporary hypoxemia in rats. A two-hit LPS stimulation induces prolonged hypoxemia and specific pulmonary injury in rats, and is therefore a more ideal approximation of ARDS in the animal model. The pathogenesis of LPS two-hit-induced ARDS is associated with an uncontrolled systemic inflammatory response and inflammatory injury. It is concluded that the rat ARDS model produced by our LPS two-hit method is more stable and reliable than previous models, and closer to the diagnostic criteria of ARDS, and better mimics the pathological process of ARDS.

Lipopolysaccharide "Two-hit" Induced Refractory Hypoxemia Acute Respiratory Distress Model in Rats / 华中科技大学学报（医学）（英德文版）

cs the pathological process of ARDS.

[Enriching of total flavonoids from Herba Leonuri with polyamide and macroporous resin].

OBJECTIVE: To study the enriching method of total flavonoid from Herba Leonuri with polyamide and macroporous resin. METHOD: Seven enriching and purifying methods were compared with the yield and purity as indexes. The method of enriching with polyamide and macroporous resin was confirmed and the process of purifying was determined by orthogonal design. RESULT: D101 resin is packed by wet method, the ratio of diameter to height is 1:7. After mixed with the extract liquids, the weight of wet resin increased to 3 times of the dry resin. Evaporated the wet resin to dryness, mixed well with a little of 95% ethanol and dry polyamide powder, evaporated them to dryness again. Elute with deionized water until the effluent being colourless, then loaded it on the macroporous adsorptive resin, elute with 50% ethanol, the volume of effluents was collected to 7 times of the column volume. The purity of total flavonoids reached to 23%, while the diversion rate from raw Herba Leonuri was to 69%. CONCLUSION: The process is simple and convenient, and the regeneration of resin is easy, which has a good application foreground.

[Effect of dahuang zhechong pill on content of amino acids in hippocampal tissue of ischemic cerebral hemorrhagic rats].

OBJECTIVE: To observe the effect of Dahuang Zhechong Pill (DZP) on the content of amino acids in hippocampal tissue of model rats of ischemic cerebral hemorrhage (ICH). METHODS: Collagenase and heparin were injected into caudate nucleus to establish ICH rat model. The content of glutamic acid (Glu), aspartic acid (Asp) and gamma-aminobutyric acid (GABA) in hippocampal tissue was detected by anti-phase high-performance liquid chromatographic fluorimetry to observe the effect of DZP on the content of amino acids in brain tissue. RESULTS: Three days after modeling, the content of Glu, Asp and GABA was significantly higher in the model group than those in the sham-operation and the normal control group (P < 0.05), for Glu 15.2926 +/- 4.2429 micromol/g vs 8.0057 +/- 1.1227 micromol/g and 8.5040 +/- 1.7794 micromol/g, for Asp 3.6384 +/- 3.1021 micromol/g vs 1.4986 +/- 0.4174 micromol/g and 1.2669 +/- 0.4695 micromol/g, and for GABA 0.3859 +/- 0.1846 micromol/g vs 0.1829 +/- 0.04665 micromol/g and 0.1770 +/- 0.0472 micromol/g. The content of Glu (9.0550 +/- 1.7195 micromol/g), Asp (1.8085 +/- 0.6862 micromol/g) and GABA (0.1993 +/- 0.0424 micromol/g) in hippocampal tissue in the DZP group was reduced significantly, as compared with that in the model group (P <0.05). CONCLUSION: DZP could reduce the release of excitatory amino acids (EAA-Glu, ASP) to balance EAA/inhibitory amino acid (IAA), so as to relieve ICH-induced brain injury.

[Effect of Chinese and western medicine integration on spinning behavior of rats with Parkinson disease].

OBJECTIVE: To observe the effect of Chinese and western medicine integration on the spinning behavior of rats with Parkinson disease. METHODS: Model of the lateral Parkinson disease was made with injection of 6-hydroxydopamine (6-OHDA) into the black substance of the right side of the brain, and the model rats were treated with madopar and Chinese herbs for nourishing the liver and kidney, clearing collaterals and detoxification. The rat's spinning behavior was observed, and was compared with the normal control group, madopar group and sham-operation group at the same time. RESULTS: Chinese and western medicine integration could obviously reduce the spinning circles of the rats. CONCLUSION: Chinese and western medicine integration can significantly improve the spinning behavior of the model rats.