Plasma levels of endothelin-1, nitric oxide,malondialdehyde and superoxide dismutase in rats exposed to infrasound

AIM: To observe the changes of NO, ET-1, SOD and MDA levels in plasma of rats exposed to infrasound. METHODS: Using infrasound (frequency: 8 Hz; sound pressure level:130 dB), the rats were exposed for 1 d， 7 d， 14 d， 21 d and 28 d, 2 h daily, then the levels of NO, ET-1, SOD and MDA were measured after exposure. RESULTS: The changes of NO levels in plasma significantly declined at 7 d and 14 d （P<0.01）, then 1, 21 and 28 d normally （P>0.05）. The changes of ET-1 levels in all groups in plasma were significantly increased （P<0.01）, mostly at 7 d, least at 14 d. The changes of SOD activity in all groups in plasma were significantly declined （P<0.01）. The changes of MDA levels in all groups in plasma were significantly increased （P<0.01）. CONCLUSIONS: Infrasouns induces changes of NO, ET-1, SOD and MDA in rat plasma, and it depends on infrasound exposure time.     

Effect of N-acetyl-L-cystein on blood pressure and endothelial function in aorta of rats exposed to chronic intermittent hypoxia

AIM: To investigate the effect of N-acetyl-L-cystein (NAC) on blood pressure and endothelial function in the aorta of the rats exposed to chronic intermittent hypoxia (CIH). METHODS: Thirty healthy male SD rats were randomly divided into 3 groups: control group, CIH group and CIH+NAC group. The systolic blood pressure (SBP) was measured with tail-cuff me-thod. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression of endothelial nitric oxide synthase (eNOS） and endothelin-1 (ET-1) in the thoracic aorta. The protein expression of eNOS in the thoracic aorta was examined by Western blotting. The levels of ET-1 in the thoracic aorta and serum were detected by radioimmunoassay. The serum nitric oxide was determined by nitric acid reduction method.The superoxide dismutase (SOD) activity in peripheral blood plasma was detected by xanthine oxidase method.The serum malondialdehyde content was detected by thiobarbituric acid method, and superoxide anion (O-·2) in thoracic aorta was determined by chemical colorimetric method. RESULTS: Compared with the control animals, CIH exposure was associated with decreased SOD level, and NAC-treated CIH animals showed recovery in SOD level. NAC treatment prevented CIH-induced hypertension as well as CIH-induced increase in MDA. The aorta eNOS mRNA and protein, and serum NO levels in CIH group were lower than those in control group, and those in NAC treatment group were higher than those in CIH group. The increases in ET-1 mRNA,ET-1 protein and O-·2 levels in the aorta, and the elevated circulating ET-1 level were also observed in CIH-exposed animals. Treatment with NAC significantly decreased the mRNA and protein levels of ET-1, the O-·2 content, and the circulating ET-1 level in CIH-exposed animals. CONCLUSION: NAC protects endothelial function and alleviates hypertension by suppressing the oxidant stress in the aorta tissues, indicating that oxidant stress may be involved in the mechanism of endothelial disorder of CIH-induced hypertension.     

Protective effect of anti-aging Klotho protein on human umbilical vein endothelial cells treated with high glucose

AIM: To study the protective effect of anti-aging Klotho protein on human umbilical vein endothelial cells (HUVECs) treated with high glucose (HG).METHODS: HUVECs were cultured in vitro, and divided into PBS control group, 5.5 mmol/L glucose group, 33.3 mmol/L glucose group, 0.1 μmol/L Klotho+33.3 mmol/L glucose group, 1 μmol/L Klotho+33.3 mmol/L glucose group, and 10 μmol/L Klotho+33.3 mmol/L glucose group. The viability of the HUVECs was measured by MTT assay. The content of malondialdehyde (MDA), and the activities of lactate dehydrogenase (LDH), superoxide dismutase (SOD) and glutathione (GSH) in cell culture supernatants were observed. The production of reactive oxygen species (ROS) in HUVECs was analyzed by flow cytometry. The levels of nitric oxide (NO), endothelin (ET-1), intercellular adhesion molecule-1 (ICAM-1) in HUVEC culture medium were detected by ELISA. The protein expression of nuclear factor-kappa B (NF-κB) in the HUVECs was determined by Western blot. RESULTS: Compared with PBS control group, 33.3 mmol/L glucose significantly decreased the HUVEC viability, increased ROS, LDH and MDA levels, reduced the activities of SOD and GSH, decreased the NO secretion, and induced the ET-1 and ICAM-1 secretion and the protein expression of NF-κB in HUVECs. When HUVECs were treated with Klotho protein at different concentrations combined with 33.3 mmol/L glucose, the cell viability was increased significantly, the ROS, LDH and MDA levels were decreased significantly, the antioxidant SOD and GSH activities were significantly increased, the secretion of NO was increased, but ET-1 and ICAM-1 releases and protein expression of NF-κB were significantly reduced.CONCLUSION: Anti-aging Klotho protein promotes the viability of HUVECs treated with HG, reduces the oxidative damage and ROS production, and restores the normal secretory function of HUVECs, thus playing a protective role in vascular endothelial cells through reducing the protein expression of NF-κB.     

Effect of external counterpulsation on asymmetry diethylarginine and endothelin levels in blood

AIM:To observe changes of serum asymmetry dimethylarginine (ADMA) and endothelin-1 (ET-1) levels,which reflect blood vessel endothelial function,after therapy of enhanced external counterpulsation(EECP) in patients who had taken percutaneous transluminal coronary angioplasty and stent.METHODS:Fifty one coronary heart disease patients (all of them had taken percutaneous tranluminal coronary angioplasty and stent) were distributed into two groups by matching them with ratio of 1∶2,17 patients in EECP group and 34 patients in control group.Both of two groups were given conventional medicine,in addition,EECP group was undertaken three courses of treatment of EECP.ADMA was detected by HPLC-fluorescence method,and ET-1 was detected by radio-immunity method.RESULTS:In EECP group ，compared with prior treatment,ADMA and ET-1 levels was obviously reduced (P<0.05).In control group,compared with prior treatment,ADMA had no significant difference (P>0.05) and ET-1 was higher (P<0.01).Compared between two groups,extent of decrease in ADMA and ET-1 levels in EECP group were more obvious than control group (P<0.01).Improvement of angina and decrease of frequency of angina was more obvious in EECP group than control group.Besides,decrease of ADMA level was positive correlation with them (r=0.85，0.70,respectively P<0.01).CONCLUSION:EECP reduces serum ADMA and ET-1 levels by increasing shear stress to vessel endothelium in coronary heart disease patients who had taken percutaneous tranluminal coronary angioplasty.It hints that EECP can improve endothelial function and provides experimental evidence for the combined treatment of coronary heart disease.     

Protective effect of procyanidin single active ingredient B2 on human endothelial progenitor cells under high glucose stimulation

AIM: To study the protective effect of procyanidin single active ingredient B2(PC-B2) on human endothelial progenitor cells(EPCs) stimulated with high glucose. METHODS: The human EPCs were isolated from peripheral blood of healthy people and identified. The EPCs were divided into control group(PBS treatment), hypertonic control group(25 mmol/L mannitol treatment), high glucose(30 mmol/L) group, and different concentrations(2, 10 and 50 mg/L) of PC-B2+30 mmol/L glucose groups. The viability of EPCs was detected by CCK-8 assay. The levels of LDH, MDA, SOD and GSH in the EPCs were detected. The changes of NO, ET-1, ICAM-1 and VCAM-1 in the EPCs cultured medium were measured by ELISA. The cell apoptotic rate and reactive oxygen species(ROS) in the EPCs were analyzed by flow cytometry. The expression of VEGF and VEGFR-2 in the EPCs were determined by Western blot. RESULTS: Compared with control group, the viability of human EPCs was decreased significantly in 30 mmol/L glucose group(P<0.05). The LDH leakage, MDA content and the releases of ET-1, ICAM-1 and VCAM-1 were induced significantly(P<0.05), but SOD and GSH activity and NO production were decreased significantly(P<0.05). The ROS and cell apoptotic rate were increased significantly(P<0.05). The expression of VEGF and VEGFR-2 in the EPCs were decreased(P<0.05). When human EPCs were treated with different concentrations of PC-B2 and 30 mmol/L glucose, the viability was obviously rebounded(P<0.05), the LDH leakage, MDA content and the releases of ET-1, ICAM-1 and VCAM-1 were decreased gradually(P<0.05), the SOD, GSH activity and NO production were increased significantly(P<0.05), the ROS and cell apoptotic rate were decreased significantly(P<0.05), and the expression of VEGF and VEGFR-2 in the EPCs was increased gradually(P<0.05).CONCLUSION: PC-B2 enhances the viability of human EPCs under high glucose condition, reduces high glucose-induced oxidative damage, restores the EPCs normal function, and reduces the releases of inflammatory cytokines and apoptosis, thus playing a protective effect on human EPCs through inducing the expression of VEGF and VEGFR-2.     

Protective effect of BNDF on vascular endothelial cells with H2O2-induced oxidative injury

AIM: To study the protective effect of brain-derived neurotrophic factor (BDNF) on vascular endothelial cells with H₂O₂-induced oxidative injury. METHODS: Human umbilical vein endothelial cells (HUVECs) were cultured in vitro, and the oxidation injury model of HUVECs was established by treatment with H₂O₂. The oxidatively injured HUVECs were cultured with different concentrations (1, 10 and 100 μg/L) of BDNF. At the same time, the control group (no injury), PBS treatment after H₂O₂ injury group and TrkB inhibitor group (with 100 μg/L BDNF and 1: 1 000 TrkB inhibitor) were also set up. The viability of the HUVECs was detected by MTT assay. The levels of LDH, MDA, SOD and GSH were measured. The releases of NO, ET-1 and ICAM-1 were analyzed by ELISA. The changes of ROS production and cell apoptosis were evaluated by flow cytometry. The protein levels of TrkB, p-TrkB, cleaved caspase-3, Bcl-2 and Bax were determined by Western blot. RESULTS: Compared with uninjured control group, in H₂O₂ oxidative injury plus PBS treatment group, the viability of the cells was decreased significantly, the LDH and MDA levels were increased significantly and the activities of SOD and GSH were decreased significantly. The NO secretion was decreased, and the ET-1 and ICAM-1 concentrations were increased significantly. The ROS content and apoptotic rate were increased significantly. The protein levels of cleaved caspase-3 and Bax were increased but Bcl-2 protein expression was decreased significantly. Compared with PBS treatment group, in H₂O₂-injured HUVECs treated with different concentrations of BDNF, the cell viability was gradually increased, the LDH and MDA levels were decreased and the activities of SOD and GSH were increased gradually. The secretion of NO was increased but ET-1 and ICAM-1 were decreased gradually. The ROS content and apoptotic rate were decreased significantly. The TrkB and p-TrkB levels were significantly increased significantly, the protein expression of cleaved-caspase 3 and Bax was decreased gradually and the Bcl-2 protein expression increased gradually. The role of BDNF was inhibited by TrkB inhibitor. CONCLUSION: BDNF protects HUVECs from oxidative injury by binding with TrkB to activate the BDNF-TrkB signaling pathways.     

Effects of Chaihu-Shugan decoction on ROCK/JNK signaling pathway and atherosclerosis in spontaneously hypertensive rats

AIM To observe the effect of Chaihu-Shugan decoction (CHSGD) on atherosclerosis in spontaneously hypertensive rats (SHR) and its possible mechanism. METHODS The male SHR (n=50) were randomly divided into model group (gavage of normal saline), compound kendir leaves (CKL) group (gavage of 0.5 g/kg CKL), and low-, medium- and high-dose CHSGD (CHSGD-L, CHSGD-M and CHSGD-H) groups (gavage of 2.5, 5 and 10 g/kg CHSGD, respectively), and another 10 male Wistar rats of the same origin were selected as normal control （NC) group (gavage of normal saline). The blood pressure was measured by intelligent noninvasive sphygmomanometer. The levels of blood lipids were measured by automatic biochemical analyzer. The expression of oxidative stress-related indexes, nitric oxide (NO), malondialdehyde (MDA) and superoxide dismutase (SOD), were detected by colorimetry. HE staining was used to detect the degree of atherosclerosis, and Western blot was used to detect the expression of Rho-associated kinase (ROCK)/c-Jun N-terminal kinase (JNK) signaling pathway-related proteins, RhoA, ROCK1 and JNK. RESULTS After 4 weeks of treatment, compared with NC group, the blood pressure, the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and MDA, and the protein expression of RhoA, ROCK1 and JNK in aortic tissues of the rats in model group were significantly increased (P<0.05), and the serum levels of high-density lipoprotein cholesterol, NO and SOD were significantly decreased (P<0.05). HE staining showed that the diameter of aortas in the rats was thickened, a large number of foam cells were formed under the endothelium, and the proliferation of smooth muscle cells was observed. Compared with model group, the blood pressure, the serum levels of TG, TC, LDL-C and MDA, and the protein expression of RhoA, ROCK1 and JNK in aortic tissues of the rats in CKL, CHSGD-L, CHSGD-M and CHSGD-H groups were significantly decreased (P<0.05), and the serum levels of NO and SOD were significantly increased (P<0.05). HE staining showed that the structure of each layer of rat aortas gradually returned to normal, the vascular cells were in good order, and the inflammatory cell infiltration was slight. Compared with CKL group, the blood pressure, the serum levels of TG, TC, LDL-C and MDA, and the protein expression of RhoA, ROCK1 and JNK in aortic tissues of the rats in CHSGD-L and CHSGD-M groups were significantly increased (P<0.05), and the serum levels of NO and SOD were significantly decreased (P<0.05). No significant difference of the above indexes between CHSGD-H group and CKL group was observed (P>0.05). CONCLUSION Chaihu-Shugan decoction may attenuate the oxidative stress response via inhibition of ROCK/JNK signaling pathway, thus alleviating the symptoms of atherosclerosis in SHR.     

Xuebijing relieves testicular ischemia/reperfusion injury in rats by activating PI3K/Akt/mTOR signaling pathway

AIM: To investigate the effect of Xuebijing on testicular ischemia/reperfusion (I/R) injury in rats and its related mechanisms. METHODS: Male Sprague-Dawley rats (n=45) were randomly divided into control group, I/R group, low-dose Xuebijing group, high-dose Xuebijing group and dexamethasone group (n=9 in each group). Except for the rats in control group, the rats in other groups underwent testicular torsion, and after the operation, the rats were treated with 0.5 mL·kg^-1·d^-1 Xuebijing, 2 mL·kg^-1·d^-1 Xuebijing and 0.5 mL·kg^-1·d^-1 dexamethasone in low-dose Xuebijing group, high-dose Xuebijing group and dexamethasone group, respectively. On the 3rd, 7th, and 14th days after treatment, the left testis in the rats of each group was taken. The histopathological changes of the testis were observed by hematoxylin-eosin staining. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), endothelin-1 (ET-1) and nitric oxide (NO) in the testicular tissue were detected by biochemical methods. The protein levels of cell cycle-related molecules, apoptosis-related proteins and PI3K/Akt/mTOR signaling pathway-related proteins were determined by Western blot. RESULTS: Xuebijing significantly attenuated the testicular damage in I/R rats, significantly increased the activity of SOD in the testis of I/R rats, reduced the content of MDA, ET-1 and NO, inhibited oxidative stress in I/R-injured tissues, mediated the protein expression of cell cycle-related factors and apoptosis-related factors, and significantly increased the protein levels of p-PI3K, p-AKT, p-mTOR and p-S6K in the testis of I/R rats (P<0.05). These effects were time-dependent and dose-dependent. CONCLUSION: Xuebijing reduces testicular I/R injury of rats by mediating the expression of cell cycle-related and apoptosis-related proteins and activating PI3K/Akt/mTOR signaling pathway in dose-dependent and time-dependent manners.     

Effects of extract of gingko biloba on the lipid metabolism and the function of macrophages from diabetic rats

AIM: To study effect of extract of ginkgo biloba (EGb) on the lipid metabolism and the function of macrophages from diabetic rats.METHODS: Sprague-Dauley rats were divided into four groups: normal control group, high-fat group, diabetic group and EGb treatment group.At the end of experiment, the rats were sacrificed, the blood glucose, blood insulin and serum lipid were measured.The activity of superoxide dismutase (SOD), content of malondialdehyde (MDA), nitric oxide (NO) in alveolar macrophages (AM) and peritoneal macrophages (PM) were assayed.In addition, peroxisome proliferator activated receptor γ (PPARγ), CD36 mRNA expression in AM was measured by RT-PCR.RESULTS: The concentration of the blood glucose, blood insulin and total cholesterol (TC), total triglycerides (TG), low-density lipoprotein- cholesterol (LDL-C) in blood increased significantly in type 2 diabetic group.The supplement of EGb decreased blood glucose, blood insulin and TC, TG, LDL-C levels.The activity of SOD decreased, while the content of NO, MDA increased in the diabetic macrophages, the activity of SOD became increased, but the content of NO and MDA decreased in EGb-treated group.The mRNA expression level of CD36 and PPARγ in alveolar macrophages from diabetic group increased, while expression level of CD36 and PPARγ mRNA in EGb treated rats continued to rise.CONCLUSIONS: EGb corrected insulin resistance and ameliorated disturbance of lipid metabolism caused by type2 diabetes in rats.Adjustment of PPARγ and CD36 mRNA expression of as well as reduction of lipid peroxidation and NO level may be involved in this process.     

Effect of non-invasive ischemic preconditioning on nitric oxide/endothelin-1 imbalance and gas exchange impairment following limb ischemia reperfusion: a clinical study

AIM: To investigate the effects of non-invasive ischemic preconditioning on nitric oxide (NO)/endothelin-1 (ET-1) imbalance and gas exchange impairment following limb ischemia reperfusion in patients undergoing unilateral lower extremity surgery with tourniquet. METHODS: Twenty-seven patients aged 25-65 years, whose tourniquets duration varied from 1 h to 1.5 h and matched American Society of Anesthesiologists Physical Status Ⅰ-Ⅱ, were randomized into two groups: a control group (n=14) and a ischemic preconditioning group (IPC group, n=13) in which patients received three cycles of 5 min of ischemia/5 min of reperfusion before tourniquet inflation. Radial arterial blood gas, plasma malondialdehyde (MDA) and NO, serum ET-1 and interleukin-6 (IL-6) were measured just before tourniquet inflation(T0), 1 h after inflation(T1), and 0.5 h(T2), 2 h(T3), 6 h(T4), 24 h(T5) after tourniquet deflation. Meanwhile NO/ET-1 ratio, alveolar-arterial oxygen gradient (PA-aDO2) and intrapulmonary shunt (Qs/Qt) were calculated. RESULTS: In control group, arterial partial pressure of oxygen (PaO2) were decreased, while PA-aDO2 and Qs/Qt were increased significantly at T4 compared to the baselines at T0 (P<0.01). Plasma NO levels and NO/ET-1 ratios decreased gradually after tourniquets deflation and statistical significances were observed at T3 (P<0.01) with a valley at T4 (P<0.01) and recovered to baselines at T5. Serum ET-1, IL-6 and plasma MDA began to increase remarkably after T3 (P<0.05 or P<0.01), peaked at T4 and dropped slightly at T5. The changes above-mentioned could be well attenuated by the application of IPC (P<0.05 or P<0.01) except PaO2 (P>0.05). CONCLUSION: Clinical application of unilateral tourniquet within safe time limit (1.5 h) may lead to limb ischemia reperfusion and further pulmonary gas exchange impairment, which could be partially attenuated by the application of IPC via alleviating NO/ET-1 imbalance.     

Renoprotective effects of sesamin on renal hypertensive and hyperlipidemic rats

AIM: To investigate the effects of sesamin on progression of renal injury in renal hypertensive and hyperlipidemic rats (RHHR). METHODS: RHHR was induced by 2K1C and high lipid baitvessel. After 7 weeks of intragastric administration with sesamin, the contents of serum creatinine (Scr), blood urea nitrogen (BUN), 24 h urinary protein excretion (UPE) were measured. In addition, the activity of total antioxidative capacity (T-AOC), superoxide dismutase (SOD), the concentrations of malondialdehyde (MDA), hydrogen peroxide (H2O2), and the nitric oxide synthase (NOS) and nitric oxide (NO) levels in renal homogenate were measured. RESULTS: Compared with the model group, seasamin (in 100 mg·kg-1 and 33 mg·kg-1 groups) evidently decreased the contents of Scr, BUN, UP and the concentration of MDA, iNOS, H2O2 in renal tissure. It also improved the levels of NO, cNOS and activity of SOD, T-AOC in renal tissure. CONCLUSION: Sesamin ameliorates hypertensive and hyperlipidemic-induced renal injury, probably by enhancing antioxidative activity, scavenging hydroxyl radical and restraining iNOS level.     

Effects of aminophylline used in the early stage of resuscitation on adenosine and vaso-active factors

AIM: To investigate the effects of aminophylline used early in resuscitation on achievement ratio of resuscitation, the concentrations of plasma norepinephrine (NE), adenosine and nitric oxide (NO), and the levels of cardiac tissue endothelin-1 (ET-1) and adenosine in rats with sudden cardiac arrest. METHODS: Sixty SD rats were randomly divided into three groups: operated control (group A), epinephrine treatment (group B), and epinephrine plus aminophylline treatment (group C). Each group had 20 rats. The concentrations of plasma NE, adenosine and NO, and the levels of cardiac tissue ET-1 and adenosine were examined in group A and 30 min after survived in group B and group C. RESULTS: The duration of circulation recovered in group C was less than that in group B, significantly (P＜0.05). The rate of return of spontaneous circulation and survival to 30 min was achieved in 75% and 70% in group C, while those were achieved in 60% and 55% in group B, respectively (P>0.05). The concentrations of plasma adenosine and NE, and the levels of cardiac tissue ET-1 and adenosine in group B and group C were higher than those in group A significantly (P<0.05). The concentration of NO in group B was also higher than that in group A (P<0.01). The levels of plasma NO and cardiac tissue ET-1 in group C was lower than those in group B significantly (P＜0.05). CONCLUSION: Aminophylline as adenosine receptor antagonist used early in resuscitation not only increases the achievement ratio of resuscitation, but also decreases the levels of ET-1 and NO, so improves after-resuscitation syndrome.     

Changes of free radicals and antioxidative system of prostate tissue in the male rats with short-term diabetes

AIM:To study the dynamic changes of free radicals and antioxidative system of prostate in rats with short-term diabetes.METHODS:48 Wistar male rats were divided into 6 groups at random (8 rats in each group). One group were injected(ip) with sodium citrate buffer to be the control group, the other 5 groups were injected (ip) with steptozocin(STZ) 60mg/kg body weight which were called diabetes group. Rats were killed after 1, 3, 5, 7 and 14 days; superoxide dismutase(SOD), malondialdehyde(MDA), glutathione(GST), glutathione s-transferase(GST), glutathione peroxidase(GSH-px), nitric oxide (NO) and nitric oxide synthase(NOS) of prostate were examined.RESULTS:The levels of MDA, GSH-px and NO in prostate homogenate of diabetes group were higher evidently as compared with control levels. The activities of SOD, GST, NOS and the levels of GSH in prostate homogenate were increased obviously after injection of STZ with control levels, then returned near to control values.CONCLUSION:The evidences mentioned above indicate that high-level of free radicals and decrease in antioxidative system rendere the prostate cells to be in oxidative stress.     

Relationship between endothelial dysfunction and renin-angiotensin-aldosterone system under the condition of fatigue stress and effect of herbs to dredge collaterals

AIM: To investigate the mechanisms and relationship among endothelial dysfunction, renin-angiotensin-aldosterone system (RAAS) and family of interleukins under the condition of excessive fatigue, by using the rats with fatigue stress. METHODS: Healthy male Wistar rats were randomly divided into control group, homocysteine (HCY) group, fatigue stress group, renshen group, shuangshen group and tongxinluo group. Radioimmunoassay was carried out to detect plasma renin activity (PRA), angiotensin II (AngⅡ), aldosterone (ALD), endothelin (ET), thromboxane-2 (TXA2), prostaglandin I2 (PGI2) in plasma and interleukin-1β, 2, 6, 10 (IL -1β, IL -2, IL -6) in sera. ELISA was used to detect NE and IL -10. The content of nitric oxide (NO) in sera was also detected. The bioinformatical analysis was used to determine the relationship between RAAS and endothelial dysfunction. RESULTS: Compared with control group, the levels of ET-1 and TXA2 in fatigue stress group were significantly increased (P<0.01, P<0.05), but the content of PGI2 and NO was significantly decreased (P<0.01, P<0.01). Compared with control group, the renin activity in plasma of animals in fatigue stress group was significantly decreased (P<0.01), the AngⅡ, IL -1β, IL -6 level was significantly increased compared with the control group (P<0.01, P<0.01), and was significantly increased compared with the HCY group (P<0.05, P<0.01, P<0.01). The NE level showed the tendency of decrease in different degree. After the intervention of three kinds of herbs to dredge collaterals, the ET-1, AngⅡ, IL -1β, IL-6 level in plasma was decreased in different degree (P<0.05, P<0.01, P<0.01), and at the same time, the contents of NO and NE level were significantly increased (P<0.05). The ALD level in tongxinluo groups was apparently higher than that in control group and the fatigue stress group (P<0.05). The bioinformatics analysis showed that Ang II and ET, IL-1; PGI2 and ALD; NO and ALD composed of three subsystems and interrelated according to the principle of optimality of complex system, and gradual change regulation was also observed in fatigue stress group. However, in control group, HCY group and tongxinluo group, the same interrelation among subsystems was not existed. CONCLUSION: In a state of long-term excessive fatigue, vascular endothelial dysfunction may be induced, and is related with renin-angiotensin-aldosterone system imbalance and serious turbulence of the autonomic dysfunction. Herbs to dredge collaterals could improve it significantly. The results suggest that bearing excessive fatigue and pressure in long-term may be the potential risk factors to induce vascular endothelial dysfunction and further result in cardio-cerebro vascular diseases, Tongluo therapy may be one of the useful ways to prevent such diseases.     

Effects of statin reloading before percutaneous coronary intervention on circulatory endothelial progenitor cells and inflammatory cytokines

AIM:To investigate the effects of atorvastatin reloading in pre-percutaneous coronary intervention (PCI) period on endothelial progenitor cell (EPC) count and inflammatory cytokine expression in the stable angina pectoris patients who had previously received long-term statin treatment. METHODS:The patients with stable angina pectoris that had received long-term statin therapy and planned to accept PCI were randomized into 3 groups: 80 mg atorvastatin 12 h and 40 mg 2 h before coronary angioplasty (80 mg reloading), pre-operatively with 40 mg/d atorvastatin for 7 d (40 mg reloading), and without atorvastatin reloading (no reloading). CD45-/CD133+/CD34+, CD45-/CD34+/KDR+ and CD45-/CD144+/KDR+ EPCs in 100 μL peripheral blood were determined by flow cytometry 1 h prior to PCI and 1 h, 6 h and 24 h after PCI. The serum concentrations of soluble intercellular adhesion molecule 1 (sICAM-1), C-reactive protein (CRP) and troponin I (TnI) were analyzed immediately prior to and 24 h after PCI. RESULTS: (1) In 80 mg reloading group, the numbers of circulating CD45-/CD133+/CD34+ and CD45-/CD34+/ KDR+ early differentiation stage EPCs 1 h and 6 h after coronary angioplasty was significantly elevated compared with those before PCI (P<0.05). (2) In control group, the serum concentrations of sICAM-1 and CRP 24 h after PCI were significantly elevated (P<0.05) compared with preoperative values. (3) The rise in serum TnI concentration from pre- to post-operation in 80 mg reloading group was lower than that in control group. CONCLUSION: The method of atorvastatin reload before PCI affects the number of EPCs in peri-operative period. High dose of atorvastatin application before PCI triggers early EPC circulation. The serum levels of post-operative inflammatory cytokine sICAM-1 as well as CRP are reduced by atorvastatin reloading before PCI.     

Effect of Chaihushugansan on pancreatic fibrosis in mice with chronic pancreatitis induced by DBTC plus ethanol and its anti-oxidation mechanism

AIM:To explore the role of superoxide dismutase (SOD) and malondialdehyde (MDA) in chronic pancreatitis (CP) induced by dibutyltin dichloride (DBTC) combined with ethanol, and the mechanisms for prevention and treatment of pancreatic fibrosis by Chaihushugansan. METHODS:The KM mice were randomly divided into control group, CP group (DBTC combined with ethanol) and Chaihushugansan group (CP+Chaihushugansan). Except for control group, the mice in other groups were intravenously injected in tail with DBTC (8 mg/kg) and drank 10% ethanol. The mice in Chaihushugansan group were administered intragastrically with Chaihushugansan (6 g·kg-1·d-1) at the following experimenal period. Before modeling and 1 week, 2 weeks, 4 weeks and 8 weeks after modeling, the mice were anesthetized and sacrificed. The activity of amylase and the content of hyaluronic acid in the serum were measured. The morphology and the degree of fibrosis in the pancreas were observed by HE staining. The activity of SOD and the level of MDA in the pancreas homogenate were analyzed. The protein of pancreas was extracted to detect the expression of type I collagen by Western blotting. RESULTS:DBTC combined with ethanol induced CP with increased serum amylase and hyaluronic acid levels， while the serum amylase and hyaluronic acid levels in Chaihushugansan group were significantly lowered (P<0.05). In 1 week, 2 weeks, 4 weeks and 8 weeks, the pancreas were obviously injured and appeared different degrees of fibrosis. The content of MDA and the expression of type I collagen in the increased significantly, but the SOD was decreased. In Chaihushugansan group, the pathological damage and the degree of fibrosis of the pancreas were improved. The level of MDA and type I collagen expression in the pancreas were significantly reduced, but the SOD was increased. CONCLUSION: The oxidative stress may take part in the development of CP. Inhibition of oxidative stress in the pancreas is one of the mechanisms that Chaihushugansan attenuates the development of CP.     

Protective effect of salidroside on rats with hypoxic pulmonary hypertension by suppressing oxidative stress

AIM: To study whether salidroside plays a protective role in hypoxia-induced pulmonary hypertension by suppressing oxidative stress. METHODS: Sprague-Dawley rats were randomly divided into 4 groups:normoxia (N) group, hypoxia for 4 weeks (H₄) group, low-dose salidroside (hypoxia for 4 weeks and treatment with salidroside at 16 mg/kg, H₄S16) group and high-dose salidroside (hypoxia for 4 weeks and treatment with salidroside at 32 mg/kg, H₄S32) group. The mean pulmonary arterial pressure (mPAP), the weight ratio of right ventricle/(left ventricle+septum)[RV/(LV+S)] and vessel wall area/vessel total area (WA/TA) were evaluated. The levels of malondialdehyde (MDA) in the serum and lung tissues were detected by colorimetric method. The levels of 8-iso-prostaglandin F_2α (8-iso-PGF_2α) in the serum and lung tissues were measured by ELISA. The activity of superoxide dismutase (SOD) in the serum was analyzed by hydroxylamine method. The expression of NAPDH oxidase 4 (NOX4) and SOD1 in the lung tissues was determined by Western blot. RESULTS: Compared with N group, the levels of mPAP, RV/(LV+S) and WA/TA in H₄ group were significantly increased, which were apparently attenuated by salidroside injection in a dose-dependent manner. Meanwhile, salidroside administration apparently decreased the levels of MDA and 8-iso-PGF_2α in the serum and lung tissues, as well as the expression of NOX4 in the lung tissues. Besides, compared with N group, the activity of SOD in the serum and the expression of SOD1 in the lung tissues in H₄ group were significantly decreased, while administration of salidroside increased the activity of SOD in the serum and the expression of SOD1 in the lung tissues in a dose-dependent manner. CONCLUSION: Salidroside protects the pulmonary vessels from remodeling and attenuates hypoxia-induced pulmonary hypertension by inhibiting oxidative stress.     

Influence of atorvastatin on scavenger receptor of monocytes/macrophages in patients with coronary heart disease

AIM: To observe activity of scavenger receptor of macrophage derived from monocytes, the change of inflammation factor in plasma (including CRP, sICAM-1, sVCAM-1) and the influence of atorvastatin on activity of scavenger receptor in patients with coronary heart disease. METHODS: 75 patients with coronary heart disease (CHD), whose plasma lipid levels were normal, were divided into three groups: stable angina pectoris, unstable angina pectoris and acute myocardia infarction. 29 healthy persons were served as control. The level of C-reactive protein (CRP), soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1) in plasma were determined in all subjects. Monocytes in peripheral blood were dissevered and cultured to transform into macrophage. The influence of atorvastatin on activity of scavenger receptor in macrophage was observed. RESULTS: The level of CRP, sICAM-1, sVCAM-1 and activity of scavenger receptor of macrophage in acute myocardia infarction group were higher than that in stable angina pectoris, unstable angina pectoris and control. Atorvastatin lowered the activity of scavenger receptor of macrophages derived from monocytes in patients with coronary heart disease. Activity of scavenger receptor of macrophages derived from monocytes in patients with coronary heart disease was correlated positively with CRP, sICAM-1 and sVCAM-1. CONCLUSION: Activity of scavenger receptor may be taken as index for monitoring the degree of active vulnerable atherosclerosis plaque. Atorvastatin may inhibit activity of scavenger receptor in macrophages derived from monocytes in patients with coronary heart disease.