[Effect of Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction on myocardial apoptosis and relevant protein expressions in miniature swine phlegm and blood stasis type coronary heart disease model].

There have been very few studies on the effect of Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction in inhibiting apoptosis in myocardial ischemial injury caused by coronary heart disease. In this experiment, Gualou Xiebai Banxia decoction combined with-Xuefu Zhuyu decoction were used to intervene the miniature swine phlegm and blood stasis type coronary heart disease model, in order to observe the effect of the combined prescription on the myocardial apoptosis and the expressions of Bcl-2, Bax, Caspase-3, Caspase-9 in the model. Totally 15 Chinese experimental miniature swine were adopted and randomly divided into the control group, the model group and the phlegm and stasis-treating group. The model group and the stasis-treating group were fed with high fat diets for two weeks, intervened with the coronary artery injury and then given drugs and high fat diets for eight weeks. The control group was fed with ordinary diets for 10 weeks, without the coronary artery injury. After the experiment, myocardia at the juncture of infracted areas were collected and made into formalin-fixed paraffin sections. The TDT-mediate dUTP nick end labeling (TUNEL) assay was used to detect the myocardial apoptosis. The immunohistochemistry (IHC) technique was applied to detect Bcl-2, Bax, Caspase-3, Caspase-9 levels in myocardial tissues. According to the findings, the apoptosis indexes (AI) for the control group, the model group and the phlegm and stasis-treating group were 0.92%, 27.68%, 17.28%, respectively. The AI of the phlegm and stasis-treating group was significantly lower than that of the model group (P < 0.01). Compared with the model group, the phlegm and stasis-treating group showed significantly higher Bcl-2 protein expression (P < 0.01) and lower Bax, Caspase-3 and Caspase-9 protein expressions (P < 0.01). In conclusion, Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction have a significant protective effect against the myocardial apoptosis in miniature swine phlegm and blood stasis type coronary heart disease model.

Long-term effect of stent coating with zedoary essential components on neointimal formation in the porcine coronary artery.

OBJECTIVE: To examine the effect of the zedoary essential component-eluting stent (ZES) on a porcine coronary neointimal formation. METHODS: ZES, sirolimus-eluting stents (SES), and bare metal stents (BMS) were randomly implanted in three different major epicardial vessels in 36 balloon-injured pigs. Coronary angiography, optical coherence tomography, and histomorphological analysis were used to determine antihyperplasia effects. RESULTS: ZES and SES had a significantly larger lumen diameter and area, and reduced diameter and area of stenosis in arteries at 30 and 90 days compared with arteries implanted with BMS (P<0.01). Histomorphometric analysis showed moderate inflammatory responses, such as infiltration of mononuclear cells, lymphocytes, and multinucleated giant cells in some arteries with SES compared with ZES (P<0.05). Injury scores were not different among the three groups at 30 and 90 days. The endothelialization score in the SES group was 2.69 ± 0.42 at 30 days and 2.83 ± 0.39 at 90 days compared with the ZES and BMS groups (both were 3.00 ± 0.00 at either 30 or 90 days, P<0.05). Well developed endothelium was observed in the ZES group, while incomplete endothelium and inflammatory cells were observed with stent struts partly naked at the vessel lumen in the SES group. CONCLUSION: The ZES inhibits neointimal hyperplasia with good endothelia coverage in the porcine balloon injury coronary model.

Hawthorn leaves flavonoids decreases inflammation related to acute myocardial ischemia/reperfusion in anesthetized dogs.

OBJECTIVE: To investigate the effects and mechanisms of hawthorn leaves flavonoids (HLF) on acute myocardial ischemia/reperfusion in anesthetized dogs. METHODS: The acute ischemia models were prepared by ligating left anterior descending (LAD) artery for 60 min. Qualified 15 male dogs were randomly divided into 3 groups with 5 in each group: blank control (treated with normal saline 3 mL/kg) group, HLF low dosage (5 mg/kg) group and high dosage (10 mg/kg) group, with an once injection through a femoral vein 5 min before reperfusion. Epicardial electrocardiogram was adopted to measure the scope and degree of myocardial ischemia. Simultaneously, neutrophil infiltration in infarct (Inf) and remote site (RS) of myocardial tissue was measured by myeloperoxidase (MPO) activity assay. The serum interleukin-1 (IL-1) and tumor necrosis factorα (TNF-α) content were quantified by radioimmuno-assay. Furthermore, expression of G protein-coupled receptor kinase 2 (GRK2) and nuclear factor κB (NF-κB) in Inf and RS tissue were detected by Western blotting technique. RESULTS: Ischemia and reperfusion increased the MPO activity and IL-1 and TNF-α content. HLF (10 and 5 mg/kg) could significantly decrease the degree and scope of myocardial ischemia; markedly inhibit the increase of MPO activity, and IL-1 and TNF-α content induced by myocardial ischemia/infarction. Furthermore, HLF increased GRK2 expression and inhibited NF-κB expression in Inf tissue. CONCLUSION: HLF could improve the situation of acute myocardial ischemia and inhibit the inflammation in anesthetized dogs, which might be due to its increasing effect on the GRK2 and NF-κB expressions.

Effects and mechanism of Weinaokang on reperfusion-induced vascular injury to cerebral microvessels after global cerebral ischemia.

OBJECTIVE: To study the effects of the Weinaokang (WNK), the active compounds extracted from Ginkgo, Ginseng, and saffron, on ischemia/reperfusion (I/R)-induced vascular injury to cerebral microvessels after global cerebral ischemia. METHODS: Male C57BL/6J mice were randomly divided into 5 groups (10 animals/group): the sham group (0.5% CMC-Na, 20 mL/kg), the I/R model group (0.5% CMCNa, 20 mL/kg), the I/R+Crocin control group (20 mg/kg), the I/R+high dose WNK group (20 mg/kg), and the I/R+low dose WNK group (10 mg/kg). Bilateral common carotid artery occlusion (BCCAO, 20 min) in mice, followed by 24 h reperfusion, was built. The generation of nitric oxide (NO), the activity of nitric oxide synthase (NOS), the phosphorylation of extracellular signal-regulated kinase1/2 (ERK1/2), and the expression of matrix metalloproteinases-9 (MMP-9) and G protein-coupled receptor kinase 2 (GRK2) in cortical microvascular homogenates were evaluated. The ultrastructural morphology of cortical microvascular endothelial cells (CMEC) was observed. RESULTS: The transient global cerebral ischemia (20 min), followed by 24 h of reperfusion, significantly promoted the generation of NO and the activity of NOS. The reperfusion led to serious edema with mitochondrial injuries in the cortical CMEC, as well as enhanced membrane GRK2 expression and reduced cytosol GRK2 expression. Furthermore, enhanced phosphorylation of ERK1/2 and decreased expression of MMP-9 were detected in cortical microvessels after I/R (20 min/24 h). As well as the positive control Crocin (20 mg/kg, 21days), pre-treatment with WNK (20, 10 mg/kg, 21 days) markedly inhibited nitrative injury and modulated the ultrastructure of CMEC. Furthermore, WNK inhibited GRK2 translocation from cytosol to the membrane (at 20 mg/kg) and reduced ERK1/2 phosphorylation and MMP-9 expression in cortical microvessels. CONCLUSION: WNK and its active compounds (Crocin) are effective to suppress I/R-induced vascular injury to cerebral microvessels after global cerebral ischemia with the target on GRK2 pathways.

[Establishment of coronary heart disease model of phlegm-stasis cementation syndrome type in mini-swines].

OBJECTIVE: To establish a disease-syndrome conjugated animal model, the mini-swine coronary heart disease (CHD) model of phlegm-stasis cementation syndrome type, by high fat diet feeding and coronary artery balloon injury. METHODS: Mini-swine were randomly divided into the control group and the model group, 6 in each group. They were fed with common forage and high fat forage respectively for 10 weeks and the coronary left anterior descending branch in the model group was injured by balloon intervention technique after 2-week feeding to establish CHD model. The model establishment and its physiopathological indices were evaluated by examinations on body mass index (BMI), blood levels of lipid, high-sensitivity C-reactive protein (hs-CRP), body surface electrocardiograph (BS-ECG), coronary angiography and pathological indices. RESULTS: BMI, total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL-C), hs-CRP, sigma-ST and N-ST indicated by BS-ECG in the model group were all higher than those in the control group respectively (P < 0.05 or P < 0.01). Coronary angiography showed coronary lumen narrowed with apparent lumen loss, showing a significant difference as compared with the control group (P < 0.01). In EVG staining, the diameters of lumen in the model group was obviously narrow with intima proliferation, also significantly different to those in the control group (P < 0.01). CONCLUSION: Disease-syndrome combined model for coronary heart disease of phlegm-stasis cementation syndrome type in mini-swine could be established by high fat diet feeding with coronary arterial injury.

[Effect of haw leaf extract and its preparation on polymorphonuclear leucocyte adhesion during HUVEC anoxia/reoxygenation injury].

OBJECTIVE: To study the effect and molecular mechanism of two haw leaf extracts, Vitexin-rhamnoside (VR) and Vitexin-glucoside (VG), and their preparation, Aoshaen injection (AI), on the polymorphonuclear leucocyte (PMN) adhesion during human umbilical vein endothelial cell (HUVEC) anoxia/reoxygenation (A/R) injury. METHODS: The cell model of A/R injury duplicated by breaking off the oxygen supplying of HUVEC for 60 min followed with reoxygenating for 30 min (phase 1) or 240 min (phase 2) was taken as the experimental objective. The effects of testing drugs (VR, VG and AI) on PMN adhesion in the model cells were measured by enzyme immunoassay, and their effects on PMN superficial adhesion molecule CD11/CD18 expression were measured by flow cytometer respectively. RESULTS: After 60 min of anoxia, HUVEC was shrunk and deformed. The adhesion between PMN and HUVEC significantly revealed at phase 1 in the model group, but it was fewer in the normal cell group, and also lesser in the groups treated with various drugs. The condition of cell adhesion revealed at phase 2 was the similar to that at phase 1. All testing drugs, VR, VG and AI, showed inhibitory effect on the cell adhesion at either phase 1 or phase 2, showing a certain dose-effect relationship. The expression of CD11/ CD18 was also inhibited by the testing drugs, and a good dose-effect relation was shown by VG and AI. CONCLUSION: At the resting condition, there are almost no expression of CD11/CD18 molecule, but it could be enhanced by incubating PMN with supernate of A/R injured HUVEC culture, and more marked at phase 1. Adding the test drugs into the supernate could inhibit the enhancing of CD11/CD18 molecule expression and reduce the PMN-HUVEC adhesion, which may be one of the molecular mechanisms of haw leaf extracts and their preparation in protecting heart against A/R injury.

[Definition and application of clinical-mimetic research in animal models with syndromes of Chinese medicine].

On the base of reviewing the current literatures concerning the animal models with syndromes of Chinese medicine and investigating thee present state of the syndrome models, the authors have put forward a definition that research of animal models with the syndromes of Chinese medicine should be combined with clinical methods, and commented the potential application. Further it was suggested that clinical methods should be used in the exploration of scientific principles and the progresses on syndromes of Chinese medicine. In addition, intervene and evaluation of Chinese herbal medicine in animal model with integrated diseases and syndromes should be emphasized to establish a platform with special properties of TCM.

[Protective effects of Shuangshen Ningxin capsule on miniature swine after myocardial ischemia by intervention].

OBJECTIVE: To study the therapeutical effects of Shuangshen Ningxin capsule on miniature swine after myocardial ischemia by intervention. METHOD: Myocardial ischemic model miniature swine induced by self-thrombus via cardiac catheter in left anteriar descending coronary artery (LAD), were administrated Shuangshen Ningxin capsule for 6 days. The changes of coronary arteriography, hemodynamics, biochemistry and pathohistology were observed. RESULT: 6 days after modeling, LAD in myocardial ischemic miniature swine was basically embolized, cardiac output (CO), cardiac index (CI), left cardiac work (LCW) and left cardiac work index (LCWI) obviously lowed, and pathohistological analysis revealed myocardial degeneration, necrosis, fibrosis and inflammatory cell infiltration. After being administered with shuangshen Ningxin capsule 6 days, the degree of self-thrombus blocked LAD reduced, hemodynamic indexes of CO, CI, LCW, LCWI and blood plasm superoxide dismutase (SOD) activity increased, and systemic vascular resistance (SVR), systemic vascular resistance index (SVRI) and malondialdehyde (MDA) content were lowed. on the same time, pathohistological degeneration and necrosis reduced. CONCLUSION: Shuangshen Ningxin capsule has anti-myocardial ischemia effect by improving cardiac muscle systolic function, increasing left cardiac work, inhibiting cardiac muscle cellular membrane lipid peroxidation.

Effects of Corocalm (shuguan capsule) on acute myocardial ischemia in anesthetized dogs.

OBJECTIVE: To investigate the effects of Corocalm (shuguan capsule) on acute myocardial ischemia in anesthetized dogs and its possible therapeutic mechanism. METHODS: The acute ischemia model was established by ligating the left anterior descending (LAD) artery. Twenty-five dogs were randomly divided into 5 groups (5 dogs in each group): the control group (treated with normal saline 3 mL/kg), the refined Guanxin Capsule group (GXC 200 mg/kg), high and low dose Corocalm groups (48.5 mg/kg for low dose group and 194.0 mg/kg for high dose group) and the Diltiazem group (5 mg/kg). The animals were treated via a single duodenal administration after the model was established. The experiments used epicardial electrocardiogram (EECG) to measure the scope and degree of myocardial ischemia. Simultaneously, the coronary blood flow (CBF) and serum activity levels of creatine phosphokinase (CK) and lactate dehydrogenase (LDH) were measured by electromagnetic flow meter and automatic biochemical analyzer respectively. The plasma endothelin (ET) content was quantified by radioimmunoassay. RESULTS: Corocalm (48.5 mg/kg and 194.0 mg/kg) significantly decreased the degree and scope of myocardial ischemia, reduced the infarct area, markedly increased the CBF, and inhibited the increase of CK and LDH activities and ET levels induced by myocardial ischemia/infarction. CONCLUSION: Corocalm could improve the state of acute myocardial ischemia and infarction in dogs. The mechanism of action might be correlated to increasing CBF, inhibiting CK and LDH activities and preventing ET release.

Cardio-protective effects of Corocalm on acute myocardial ischemia/reperfusion injury in rats.

OBJECTIVE: To investigate the cardio-protective effects of Corocalm on acute myocardial ischemia in rats, and to explore its possible therapeutic mechanisms. METHODS: The acute ischemic model was prepared by ligating the left anterior descending (LAD) coronary artery in rats. The animals were divided into 6 groups, 8 in each group. The sham operated group underwent heart exposure without ligation and were treated with normal saline 3 ml/kg, while the other 5 groups, the model groups, consisted of acceptable acute ischemic model rats and were also treated with normal saline, with the Guanxin Capsule (GXC) group treated with refined GXC, 600 mg/kg, the low and high dose Corocalm groups treated with 85 mg/kg and 340 mg/kg of Corocalm respectively, and the Diltiazem group, treated with Diltiazem 5 mg/kg, with all the tested drugs prepared with normal saline into equal volume (3 ml/kg) and administrated once via duodenum 10 min before ligation. Myocardial infarction area was determined by the quantitative histological assay with nitroblue tetrazolium (N-BT) stain. And the levels of creatine phosphokinase (CK), lactate dehydrogenase (LDH), malondialdehyde (MDA) content, and the activity of superoxide dismutase (SOD) in serum were measured by biochemical assay and spectrophotometry respectively. Besides, the blood viscosity in another 50 rats was determined, who received for 7 successive days oral administration with different concentration of Corocalm or aspirin. RESULTS: It showed that low and high dose Corocalm could significantly reduce the infarction area, inhibit the increase of serum CK, LDH activity and MDA content, and enhance the SOD activity after ischemia/reperfusion. The whole blood viscosity at different shear rates in rats treated with high dose Corocalm was significantly lower than those treated with normal saline (P < 0.05). CONCLUSION: Corocalm has favourable protective effects on heart in ischemic condition, the effect of which might be through its actions in inhibiting CK and LDH activity, scavenging oxygen free radicals, and lowering blood viscosity.

[Effects of shuangshen ningxin capsule on mini-porcine myocardial ischemia model induced by thrombosis via cardiac catheterization intervention].

OBJECTIVE: To investigate the establishment of mini-porcine myocardial ischemia (MI) model induced by thrombosis via cardiac catheterization intervention, and observe the anti-MI action of Shuangshen Ningxin Capsule (SNC). METHODS: MI model of Chinese mini-porcine was established by injection of self-thrombus in left anterior descending coronary artery (LAD) with guiding catheter through carotid artery. The effect of SNC on MI was evaluated comprehensively by analyzing the parameters as coronary embolism, 30 dots surface mapping electrocardiogram and quantitative histology, etc. RESULTS: LAD in the model animals was basically embolized 6 days after modeling, showing increase of amplitude and dot of ST segment elevation. After being administered with SNC for 6 days, the self-thrombus blocked LAD was recanalized and the degree and extent of MI reduced. CONCLUSION: It is the first time to successfully establish MI model of Chinese mini-procine by thrombosis via cardiac catheterization intervention. SNC has anti-MI effect.

[Protective effects of saponines of stem and leaf of Panax notoginseng on acute myocardial ischemia in anaesthetic dogs].

OBJECTIVE: To study the protective effects of saponines of stem and leaf of Panax notoginseng (PNSSL) on acute myocardial ischemia in anaesthetic dogs. METHOD: The acute ischemia models were made by ligation of left anterior descending (LAD) artery. The myocardial blood flow (MBF) was determined by ultrasonic doppler. The experiments adopted epicardiogram mapping to measure the scope and degree of myocardial ischemia, quantitative histologic assay (nitroblue tetrazolium, N-BT stain) to determine the size of myocardial infarction. And the endothelin (ET) and thromboxane B2 (TXB2) were measured by radioimmunological assay. RESULT: PNSSL was showed to obviously alleviate the degree of myocardial ischemia (sigma-ST) and narrow the ischemic area indicated by N-BT staining. In addition, PNSSL could increase the MBF of ischemia section. And the treatment could inhibit the ET and TXB2 release induced by ischemia and infarction. CONCLUSION: PNSSL demonstrated to attenuate the damage subjected to myocardial ischemia and infarction, which may be due to its function of inhibiting the ET and TXA2 release, increasing the MBF, and then improving the damaged cardiac function.