Salvianolate increases heat shock protein expression in a cerebral ischemia-reperfusion injury model

Stroke remains a worldwide health problem. Salvianolate exerts a protective effect in various mi- crocirculatory disturbance-related diseases, but studies of the mechanisms underlying its protective action have mainly focused on the myocardium, whereas little research has been carried out in brain tissue following ischemia-reperfusion. We assessed the neuroprotective effects of salvianolate in a rat model of cerebral ischemia-reperfusion injury induced using the suture method. At onset and 24 and 48 hours after reperfusion, rats were intraperitoneally injected with salvianolate （18 mg/kg） or saline. Neurological deficit scores at 72 hours showed that the neurological functions of rats that had received salvianolate were significantly better than those of the rats that had received saline. 2,3,5-Triphenyltetrazolium chloride was used to stain cerebral tissue to determine the extent of the infarct area. A significantly smaller infarct area and a significantly lower number of apoptotic cells were observed after treatment with salvianolate compared with the saline treatment. Expression of heat shock protein 22 and phosphorylated protein kinase B in ischemic brain tissue was significantly greater in rats treated with salvianolate compared with rats treated with saline. Our findings suggest that salvianolate provides neuroprotective effects against cerebral ischemia-reperfusion injury by upregulating heat shock protein 22 and phosphorylated protein kinase B expression.

Protection of Salvianolate against Atherosclerosis via Regulating the Inflammation in Rats

Inflammation plays an essential role in the pathophysiology of atherosclerosis. Our study was aimed to investigate whether salvianolate, a novel water-soluble phenolic compound of Danshen, alleviates atherosclerosis via regulating the inflammation in rats. High fat diet feeding plus vitamin D3 injection was used to induce atherosclerosis in rats. Salvianolate （60, 120 or 240 mg/kg） or placebo was given to atherosclerotic rats. The plasma lipids, interleukin 6 （IL-6） and C reactive protein （CRP） were measured by ELISA. CD4＋CD25＋Foxp3＋ cells were determined by flow cytometry. Histological changes were examined by hematoxylin and eosin staining. The results showed that the levels of plasma IL-6 and CRP were elevated in the rats fed on high fat diet, and the histological analysis demonstrated the successful establishment of atherosclerosis models. Treatment with salvianolate alleviated the atherosclerotic process and decreased the levels of plasma IL-6 and CRP. Also the number of CD4＋CD25＋Foxp3＋ cells was increased in salvianolate-treated rats. It was concluded that salvianolate could treat atherosclerosis via modulating the inflammation at cytokine and cell levels.

LapRLSR for NIR spectral modeling and its application to online monitoring of the column separation of Salvianolate

A novel near infrared （NIR） modeling method-Laplacian regularized least squares regression （LapRLSR） was presented,which can take the advantage of many unlabeled spectra to promote the prediction performance of the model even if there are only few calibration samples. Using LapRLSR modeling, NIR spectral analysis was applied to the online monitoring of the concentration of salvia acid B in the column separation of Salvianolate. The results demonstrated that LapRLSR outperformed partial least squares （PLS） significantly, and NIR online analysis was applicable.

Changes in plaque stability, cell apoptosis and coagulation indexes after patients with unstable angina pectoris accept adjuvant salvianolate therapy

Objective:To investigate the effect of adjuvant salvianolate therapy on plaque stability, cell apoptosis and coagulation indexes in patients with unstable angina pectoris.Methods: 92 patients with unstable angina pectoris treated in our hospital between May 2011 and August 2015 were collected, and after the treatment process and auxiliary examination results were retrospectively analyzed, they were divided into the control group (n=45) who accepted conventional treatment and the observation group (n=47) who accepted adjuvant salvianolate treatment. Before and after treatment, diasonograph was used to evaluate the plaque stability parameters of two groups of patients;ELISA was used to detect apoptosis-related molecule levels;immunoturbidimetry was used to detect blood coagulation indexes.Results: Before treatment, differences in plaque stability parameters, cell apoptosis molecules and coagulation indexes were not statistically significant between two groups of patients (P>0.05). After treatment, the plaque stability parameters plaque thickness, enhanced intensity, rise time and time to peak of observation group were significantly lower than those of control group (P<0.05);serum sFas, sFasL, fibrinogen (Fib), platelet (PLT), and D-Dimer (D-D) levels of observation group were significantly lower than those of control group while Bcl-2, prothrombin time (PT) and activated partial thromboplastin time (APTT) levels were significantly higher than those of control group (P<0.05).Conclusions: Adjuvant salvianolate treatment can increase the plaque stability, also inhibit myocardial cell apoptosis and improve the coagulation function in patients with unstable angina pectoris.

Effect of salvianolate therapy on peripheral blood Bcl-2, BAX and Caspase-3 expression in patients with cerebral ischemic stroke and their correlation with neural function

Objective:To study the peripheral blood Bcl-2, BAX and Caspase-3 expression in patients with cerebral ischemic stroke before and after salvianolate therapy and their correlation with neural function.Methods:A total of 89 patients with cerebral ischemic stroke who received salvianolate therapy in Neurology Department of Zigong Fourth People's Hospital between May 2014 and April 2016 were studied. Before treatment as well as 2 weeks and 4 weeks after treatment, the peripheral blood apoptotic molecules Bcl-2, BAX and Caspase-3 expression as well as serum neural cytokine and oxidative stress product levels were determined respectively. Results:2 weeks and 4 weeks after treatment, peripheral blood Bcl-2 mRNA expression as well as serum BDNF, NGF, VEGF and IGF-1 levels were significantly higher than those before treatment while BAX and Caspase-3 mRNA expression as well as serum MDA, 8-OHdG and 8-iso-PGF2α levels were significantly lower than those before treatment;4 weeks after treatment, peripheral blood Bcl-2 mRNA expression as well as serum BDNF, NGF, VEGF and IGF-1 levels was significantly higher than those 2 weeks after treatment while BAX and Caspase-3 mRNA expression as well as serum MDA, 8-OHdG and 8-iso-PGF2α levels were significantly lower than those 2 weeks after treatment. Peripheral blood Bcl-2 mRNA expression was positively correlated with serum BDNF, NGF, VEGF and IGF-1 levels, and negatively correlated with serum MDA, 8-OHdG and 8-iso-PGF2α levels;peripheral blood BAX and Caspase-3 mRNA expression were negatively correlated with serum BDNF, NGF, VEGF and IGF-1 levels, and positively correlated with serum MDA, 8-OHdG and 8-iso-PGF2αlevels.Conclusion:Salvianolate treatment of cerebral ischemic stroke can increase the anti-apoptotic molecule expression and inhibit the pro-apoptotic molecule expression to improve the neural function.

Role of CD36 in central nervous system diseases

CD36 is a highly glycosylated integral membrane protein that belongs to the scavenger receptor class B family and regulates the pathological progress of metabolic diseases.CD36 was recently found to be widely expressed in various cell types in the nervous system,including endothelial cells,pericytes,astrocytes,and microglia.CD36 mediates a number of regulatory processes,such as endothelial dysfunction,oxidative stress,mitochondrial dysfunction,and inflammatory responses,which are involved in many central nervous system diseases,such as stroke,Alzheimer’s disease,Parkinson’s disease,and spinal cord injury.CD36 antagonists can suppress CD36 expression or prevent CD36 binding to its ligand,thereby achieving inhibition of CD36-mediated pathways or functions.Here,we reviewed the mechanisms of action of CD36 antagonists,such as Salvianolic acid B,tanshinone IIA,curcumin,sulfosuccinimidyl oleate,antioxidants,and small-molecule compounds.Moreover,we predicted the structures of binding sites between CD36 and antagonists.These sites can provide targets for more efficient and safer CD36 antagonists for the treatment of central nervous system diseases.

A Prospective Randomized Multicenter Controlled Trial on Salvianolate for Treatment of Unstable Angina Pectoris in A Chinese Elderly Population

Objective: To evaluate the efficacy and safety of salvianolate in elderly patients with unstable angina pectoris(UAP). Methods: A prospective double-blind randomized placebo-controlled multicenter trial in elderly patients with UAP from 13 third-grade class-A hospitals in China was performed. A total of 318 patients were randomly allocated in a 1:1 ratio to an experimental group(160 patients) and a control group(158 patients). The experimental group was treated with salvianolate for 14 days on the basis of conventional medicine, and the control group was given a placebo for 14 days with the same criteria. Follow-up was lasted 28 days in both groups. The primary endpoint was biweekly frequency of angina pectoris attacks. The secondary endpoints included biweekly dosage of nitroglycerin, the Seattle Angina Questionnaire, angina pectoris severity and duration, myocardial injury markers, high-sensitivity C-reactive protein(hs-CRP) and N-terminal pro-B-type natriuretic peptide(NT-proBNP), as well as major adverse cardiovascular events(MACEs). Safety was assessed according to adverse events and serious adverse events. Results: Baseline characteristics were similar between treatment groups. Compared with those in the control group, the frequency of biweekly angi na attacks(2.92 vs. 4.08, P =0.025), th e biweekly dosage of nitroglycerin, as well as the severity and duration of angina attacks(P <0.01) were reduced by salvianolate. The Seattle Angina Questionnaire score was also significantly improved in the experimental group than in the control group(P <0.05). No significant differences were observed between the two groups with respect to the incidence of MACEs. Salvianolate was well tolerated. Conclusions: Salvianolate appear to have efficacy and well tolerated for elderly patients with UAP.

Salvianolate Reduces Murine Myocardial Ischemia and Reperfusion Injury via ERK1/2 Signaling Pathways in vivo

Objective： To analyze the effects of salvianolate on myocardial infarction in a murine in vivo model of ischemia and reperfusion （I/R） injury. Metheds： Myocardial I/R injury model was constructed in mice by 30 min of coronary occlusion followed by 24 h of reperfusion and pretreated with salvianolate 30 min before I/R （SAL group）. The SAL group was compared with SHAM （no I/R and no salvianolate）, I/R （no salvianolate）, and ischemia preconditioning （IPC） groups. Furthermore, an ERK1/2 inhibitor PD98059 （1 mg/kg）, and a phosphatidylinositol-3-kinase （PI3-K） inhibitor, LY294002 （7.5 mg/kg）, were administered intraperitoneal injection （i.p） for 30 min prior to salvianolate, followed by I/R surgery in LY and PD groups. By using a double staining method, the ratio of the infarct size （IS） to left ventricle （LV） and of risk region （RR） to LV were compared among the groups. Correlations between IS and RR were analyzed. Western-blot was used to detect the extracellular signal-regulated kinase 1/2 （ERK1/2） and protein kinase B （AKT） phosphorylation changes. Results： There were no significant differences between RR to LV ratio among the SHAM, I/R, IPC and SAL groups （P〉0.05）. The SAL and IPC groups had IS of 26.1% ± 1.4% and 22.3% ±2.9% of RR, respectively, both of which were significantly smaller than the I/R group （38.5% ± 2.9% of RR, P〈0.05, P〈0.01, respectively）. Moreover, the phosphorylation of ERK1/2 was increased in SAL group （P〈0.05）, while AKT had no significant change. LY294002 further reduced IS, whereas the protective role of salvianolate could be attenuated by PD98059, which increased the IS. Additionally, the IS was not linearly related to the RR （r=0.23, 0.45, 0.62, 0.17, and 0.52 in the SHAM, I/R, SAL, LY and PD groups, respectively）. Conclusion： Salvianolate could reduce myocardial I/R injury in mice in vivo, which involves an ERK1/2 pathway, but not a PI3-K signaling pathway.

Protective Effects of Salvianolate on Myocardial Injury or MyocardiaMnfarction after Elective Percutaneous Coronary Intervention in NSTE-ACS Patients:A Randomized Placebo-Controlled Trial

Objective:To evaluate the protective effects of salvianolate on percutaneous coronary intervention(PCI)related myocardial injury or myocardial infarction after elective PCI in non-ST-segment elevation acute coronary syndrome(NSTE-ACS)patients.Methods:A total of 149 patients with NSTE-ACS who underwent elective PCI were enrolled.The patients were randomly allocated in a 1:1 ratio to the salvianolate group(74 cases)or the control group(75 cases).After exclusion criteria of coronary angiography,60 patients with PCI therapy remained in the salvianolate group and 68 in the control group.The incidence and the severity of PCI related myocardial injury or myocardial infarction,in addition to major adverse cardiac events(MACEs)during 1 year follow-up after PCI were studied between the two groups.Multivariate logistic regression analysis was used to determine the independent factors for PCI related myocardial injury or myocardial infarction after elective PCI.Results:Compared with the control group,salvianolate treatment reduced the incidence of PCI related severe myocardial injury or myocardial infarction(11.7%vs.26.5%,P=0.035).The rate of MACEs or all-cause death within 1 month or 1 year after the procedure was not significantly different between the two groups.Conclusions:Periprocedural treatment with salvianolate reduces the incidence of PCI related severe myocardial injury or myocardial infarction,although it does not influence clinical prognosis.

Protection of Salvianolate Lyophilized Injection combined with Xueshuantong Injection (Lyophilized) against focal cerebral ischemia/reperfusion injury in rats through suppression of inflammatory response

Objective： Inflammatory reactions induced by microglia in the brain play an important part in the pathogenesis of focal cerebral ischemia/reperfusion （I/R） injury, resulting in neuronal death. Salvianolate Lyophilized Injection （SLI） and Xueshuantong Injection （Lyophilized） （XST）, which have been widely used in the treatment of acutely cerebral infarction clinically in China, exhibit various biological activities. In this study, the neuroprotective properties of SLI combined with XST in a rat model of middle cerebral artery occlusion- reperfusion （MCAO/R） were investigated. Methods： In this study, male Wistar rats were subjected to 1.5h of middle cerebral artery occlusion followed by reperfusion for 24 h. The rats were randomly divided into the following six groups： normal group （NOR）, model group （MOD）, SLI group （21 mg/kg, SLI）, ）（ST group （100 mg/kg, ）（ST）, SLI combined with XST （XST 100 mg/kg ＋ SLI 21 mg/kg, 1X1S）, and Edaravone （as a positive control drug, 6 mL/kg, EDI）, once a day for 3 d. The neuronal injury, the expression of glial fibrillary acidic protein （GFAP） and ionized calcium binding adaptor molecule 1 （IBA-1）, and the changes of pro-inflammatory mediators interleukin- 6 （IL-6）, tumor necrosis factor alpha （TNF-α） and anti-inflammatory mediator interleukin-10 （IL-10） were observed. Results： 1X1S treatment significantly increased the number of neuron, compared with the MOD group, SH group and XST group. Gliosis （GFAP and IBA-1） and expression of pro-inflammatory mediators IL-6 and TNF-a were significantly reduced. Meanwhile, 1XIS significantly increased the expression of anti- inflammatory mediator IL-10 in the brains of MCAO/R rats, compared with the MOD group, SLI and XST groups. SLI and XST also remarkably down-regulated the expression of IL-6 and TNF-α compared with the MOD group. Conclusions： This study shows that SLI combined with XST （1X1S） can protect cerebral ischemia- reperfusion injury due to its anti-inflammatory property, and may provide a potential promising new therapeutic strategy for acute ischemic stroke.

Salvianolate Reduces Glucose Metabolism Disorders in Dimethylnitrosamine-Induced Cirrhotic Rats

Objective： To evaluate the preventive effect of salvianolate （Sal B） on glucose metabolism disorders of dimethylnitrosamine （DMN）-induced cirrhotic rats. Methods： Fiffy-five Wistar rats were randomly divided into a control group （n=10） and a cirrhotic group （n=45） according to a random number table. Liver cirrhosis was induced by intraperitoneal administration of DMN. The cirrhotic rats were divided into model, Sal B and metformin groups （n=15）, respectively. Rats in the model group were given saline, two treatment groups were given Sal B （50 mg/kg）, metformin （150 mg/kg） respectively for 28 consecutive days, while rats in the control group were injected 0.9% saline with same volume of vehicle. Body weight was measured everyday. Insulin sensitivity was determined by euglycemic hyperinsulinemic clamp. Organ index, glucose tolerance test （OGTT）, and fasting plasma glucose （FPG）, fasting insulin （FINS）, hepatic glycogen, hydroxyproline （HYP） and liver function were detected at the end of the treatment. Area under the curve （AUC） for OG3-1 was calculated. Liver and pancreas histology were determined by histopathological examination with hematoxylin and eosin staining （HE）, Sirius Red staining and Masson＇s trichrome staining, respectively. Hepatic expression of α-smooth muscle actin （oL-SMA） and collagen （Col I ） were evaluated by immunohistochemical staining. Results： Compared with the model group, Sal B significantly increased body and liver weight, liver-body ratio, glucose infusion rate （GIR）, FPG, FINS levels and hepatic glycogen at the end of administration （P〈0.05 or P〈0.01）. Meanwhile, Sal B significantly decreased AUC for OGI-I＇, spleen weight, spleen-body ratio, aminotransferase and HYP level （P〈0.05 or P〈0.01）. Sal B was also effective in alleviating necrosis of liver tissue, suppressing fibrosis progression and inhibiting the expression of oL-SMA and Col I in liver. Compared with the metformin group, Sal B had advantages in ameliorating FPG, hepatic glycogen, spleen weight, organ index, liver function and cirrhosis （P〈0.05）. Metformin increased insulin sensitivity more potently than Sal B （P〈0.05）. Conclusions： Sal B could improve glucose metabolism in cirrhotic rats by protecting hepatic glycogen reserve, increasing insulin sensitivity, and alleviating pancreatic morphology abnormalities. Sal B was clinically potential in preventing glucose metabolism anomalies accompanied with cirrhosis.

Determination of Salvianolic Acid B in Yiqi Huayu Prescription by HPLC

[Objectives]To determine the content of salvianolic acid B in Yiqi Huayu Prescription by HPLC.[Methods]The chromatographic column was ZORBAX Eclipse Plus C 18(4.6 nm×250 nm,5μm);the mobile phase was acetonitrile-0.1%phosphoric acid(21:79),the detection wavelength was 286 nm,the column temperature was 30℃,and the flow rate was 1.0 mL/min.A method for determination of salvianolic acid B in Yiqi Huayu Prescription was established.[Results]The linear relationship of salvianolic acid B was good in the range of 0.0214-0.4064 mg/mL.The regression equation was Y=5995.98984 X-0.07332,r=0.9999.The average recovery rate was 98.88%(RSD=1.6%).[Conclusions]The method is reliable,accurate and specific,and can be used for the determination of salvianolic acid B in Yiqi Huayu Prescription.

Study on Optimization of Ethanol Reflux Extraction of Phenolic Acids from Salvia miltiorrhiza

The extraction technology of phenolic acid compounds from Salvia miltiorrhiza by ethanol reflux was studied. In this experiment, salvianolic acid B standard was used to make the standard curve. Single factor experiment and orthogonal experiment were used to study the extraction of different ethanol concentrations, reflux times and material-to-liquid ratios. The OD value of salvianolic acid compounds was measured with a spectrophotometer. The extraction rate of phenolic acid compounds under different extraction conditions was calculated through a regression equation, so as to obtain the optimal conditions for the ethanol reflux extraction process of Salvia miltiorrhiza. The experimental data can provide a reference for the ethanol reflux extraction process of salvianolic acids in the industry. According to the experiment, the extraction rate of phenolic acids in Salvia miltiorrhiza was the highest when the ethanol concentration was 60%, the reflux time was 1.5 hours, and the ratio of material-to-liquid was 1:10.

Effect of salvianolic acid B-loaded mesoporous silica nanoparticles on myocardial ischemia-reperfusion injury

Background:Currently,no drugs can specifically improve clinical cardiac ischemia-reperfusion injury or the prognosis of hemodialysis.Salvianolic acid B(SalB)is a widely used cardiac protectant;however,its clinical application is limited by its low oral bioavailability and poor intestinal absorption.The exploration of its preparation and clinical applications has become a research hotspot in recent years.Methods:To determine whether mesoporous silica nanoparticles(MSNs)efficiently delivered SalB to the heart and SalB@MSNs-RhB reduced myocardial ischemia-reperfusion injury,we constructed a myocardial ischemia-reperfusion male rat model,hypoxia/reoxygenation cardiomyocytes,and treated them with SalB@MSNs-RhB.Results:SalB@MSNs-RhB showed improved bioavailability,therapeutic effect,heightened JAK2/STAT3-dependent pro-survival signaling,and antioxidant responses,thereby protecting cardiomyocytes from ischemia-reperfusion injury-induced oxidative stress and apoptosis.Conclusion:This use of SalB-loaded nanoparticles and investigation of their mechanism of action may provide a new strategy for treating cardiomyocytes.Thus,hypoxia/reoxygenation promotes the clinical application of SalB.

Effect and mechanism of salvianolic acid B on the myocardial ischemiareperfusion injury in rats

Objective:To investigate the effect of salvianolic acid B on rats with myocardial ischemiareperfusion injury.Methods:SD rats were randomly divided into five groups(n=10 in each group):A sham operation group,B ischemic reperfusion group model group,C low dose salvianolic acid B group,D median dose salvianolic acid B group,E high dose salvianolic acid B group.One hour after establishment of the myocardial ischemia-reperfusion model,the concentration and the apoptotic index of the plasma level of myocardial enzymes(CTnⅠ,CKMB),SOD,MDA,NO,ET were,measured.Heart tissues were obtained and micro-structural changes were observed.Results:Compared the model group,the plasma CTnⅠ,CK-MB,MDA and ET contents were significantly increased,NO,T-SOD contents were decreased in the treatment group(group C,D,and E)(P<0.05);compared with group E,the plasma CTnⅠ,CKMB,MDA and ET levels were increased,the NO,T-SOD levels were decreased in groups C and D(P<0.05).Infarct size was significantly reduced,and the myocardial ultrastructural changes were improved significantly in treatment group.Conclusions:Salvianolic acid B has a significant protective effect on myocardial ischemia-reperfusion injury.It can alleviate oxidative stress,reduce calcium overload,improve endothelial function and so on.

Effects of salvianolic acid-A on NIH/3T3 fibroblast proliferation,collagen synthesis and gene expression

AIM To investigate the mechanisms ofsalvianolic acid A（SA-A）against liver fibrosisin vitro.METHODS NIH/3T3 fibroblasts were culturedroutinely,and incubated with 10-4mol/L-10-7mol/L SA-A for 22 h.The cell viability wasassayed by[3H]proline incorporation,cellproliferation by[3H]TdR incorporation,cellcollagen synthetic rate was measured with[3H]proline impulse and collagenase digestionmethod.The total RNA was prepared from thecontrol cells and the drug treated cellsrespectively,and α（1）I pro-collagen mRNAexpression was semi-quantitatively analyzedwith RT-PCR.RESULTS 10-4mol/L SA-A decreased cellviability and exerted some cytotoxiciy,while10-5mol/L-10-7mol/L SA-A did not affect cellviability,but inhibited cell proliferationsignificantly,and 10-6mol/L SA-A had the besteffect on cell viability among theseconcentrations of drugs.10-5mol/L-10-6mol/LSA-A inhibited intracellular collagen syntheticrate,but no significant influence on extracellularcollagen secretion.Both 10-5mol/L and10-6mol/L SA-A could decrease α（1）I pro-collagen mRNA expression remarkably.CONCLUSION SA-A had potent action againstliver fibrosis.It inhibited NIH/3T3 fibroblastproliferation,intracellular collagen syntheticrate and type I pro-collagen gene expression,which may be one of the main mechanisms of thedrug.

Effects of salvianolic acid B on liver mitochondria of rats with nonalcoholic steatohepatitis

AIM: To investigate the effects of salvianolic acid B(Sal B) on the morphological characteristics and functions of liver mitochondria of rats with nonalcoholic steatohepatitis(NASH).METHODS: A total of 60 male Sprague-Dawley rats were randomly divided into three groups:(1) a normal group fed a normal diet;(2) an NASH model group; and(3) a Sal B-treated group fed a high-fat diet. Two rats from each group were executed at the end of the 12 th week to detect pathological changes. The rats in the Sal B-treated group were gavaged with 20 m L/kg Sal B(1 mg/m L) daily. The model group received an equal volume of distilled water as a control. At the end of the 24 th weekend, the remaining rats were executed. Serum biochemical parameters and liver histological characteristics were observed. Malondialdehyde(MDA) and superoxide dismutase(SOD) in the liver were determined. Protein expression of Cyt C and caspase-3 was determined by immunohistochemistry. The m RNA transcripts of mitofusin-2(Mfn2) and NF-κB in the liver tissue were detected by real-time PCR. Mitochondrial membrane potential was detected using a fluorescence spectrophotometer. Mitochondrial respiratory function was detected using a Clark oxygen electrode.RESULTS: The model group showed significantly higher ALT, AST, TG, TC and MDA but significantly lower SOD than the normal group. In the model group,the histological characteristics of inflammation and steatosis were also evident; mitochondrial swelling and crest were shortened or even disappeared. Cyt C(18.46 ± 1.21 vs 60.01 ± 3.43, P < 0.01) and caspase-3 protein expression(30.26 ± 2.56 vs 83.31 ± 5.12, P < 0.01) increased significantly. The m RNA expression of NF-κB increased(0.81 ± 0.02 vs 0.91 ± 0.03, P < 0.05), whereas the m RNA expression of Mfn2 decreased(1.65 ± 0.31 vs 0.83 ± 0.16, P < 0.05). Mitochondrial membrane potential also decreased and breathing of rats was weakened. Steatosis and inflammation degrees in the treatment group were significantly alleviated compared with those of the model group. In the treatment group, mitochondrial swelling was alleviated. Cyt C(60.01 ± 3.43 vs 30.52 ± 2.01, P < 0.01) and caspase-3 protein expression(83.31 ± 5.12 vs 40.15 ± 3.26, P < 0.01) significantly decreased. The m RNA expression of NF-κB also decreased(0.91 ± 0.03 vs 0.74 ± 0.02, P < 0.01), whereas the m RNA expression of Mfn2 increased(0.83 ± 0.16 vs 1.35 ± 0.23, P < 0.01). Mitochondrial membrane potential increased and respiratory function was enhanced. CONCLUSION: Sal B can treat NASH by protecting the morphological characteristics and functions of liver mitochondria, regulating lipid metabolism, controlling oxidative stress and lipid peroxidation and inhibiting apoptosis.

Effect of salvianolic acid B on Smad3 expression in hepatic stellate cells

BACKGROUND: Salvianolic acid B (SA-B), one of water soluble compounds derived from Radix salviae miltiorrhizae, had good action against liver fibrosis of patients with chro- nic hepatitis. Hepatic stellate cells (HSCs) is the cellular re- source for liver fibrogenesis, while transforming growth factor-β1 (TGF-β1) is most potent fibrogenic factor. In this study we investigated the mechanism of SA-B action against liver fibrosis relating to the interference with TGF- β1 signaling at HSC. METHODS: Hepatic stellate cells (HSCs) were isolated, cultured, and incubated with SA-B. The TGF-β1 content in the supernatant of subcultured HSCs was assayed with ELISA. Type I collagen and Smad3 protein in TGF-β1-sti- mulated primarily cultured HSCs for 4 days were detected by Western blot. RESULTS: TGF-β1 secreted in activated HSCs was more than in primary HSCs, and SA-B significantly decreased TGF-β1 secretion in activated HSCs. TGF-β1 increased the expression of type I collagen and Smad3 protein in d4 pri- mary HSCs, while SA-B inhibited their expression. CONCLUSIONS: SA-B inhibits TGF-β1 secretion in activa- ted HSCs and counteracts the expression of TGF-β1 stimu- lated type I collagen and Smad3. These actions are associat- ed with the effect of SA-B on liver fibrosis.

Protective actions of salvianolic acid A on hepatocyte injured by peroxidation in vitro

INTRODUCTION Salvianolic radix,one of the most commonly usedtraditional Chinese herbs,was widely studied aboutits actions against liver injury and fibrosis,and wasone of the focuses of recent research.Salvianolic acid-A(SA-A)was an aqueous solublecomponent of Salvianolic radix.In our

Effects of salvianolic acid B on in vitro growth inhibition and apoptosis induction of retinoblastoma cells

AIM: To observe the effects of salvianolic add B (SalB) on in vitro growth inhibition and apoptosis induction of retinoblastoma HXO-RB44 cells. METHODS: The effects of SalB on the HXO-RB44 cells proliferation in vitro were observed by MTT colorimetric method. The morphological changes of apoptosis before and after the treatment of SalB were observed by Hoechst 33258 fluorescent staining method. Apoptosis rate and cell cycle changes of HXO-RB44 cells were detected by flow cytometer at 48 hours after treated by SalB. The expression changes of Caspase-3 protein in HXO-RB44 cells were detected by Western Blot. RESULTS: SalB significantly inhibited the growth of HXO-RB44 cells, while the inhibition was in a concentration-and time-dependent manner. The results of fluorescent staining method indicated that HXO-RB44 cells showed significant phenomenon of apoptosis including karyorrhexis, fragmentation and the formation of apoptotic bodies, etc. after 24, 48 and 72 hours co-culturing of SalB and HXO-RB44 cells. The results of flow cytometer showed that the apoptosis rate and the proportion of cells in S phase were gradually increased at 48 hours and 72 hours after treated by different concentrations of SalB. Western Blot strip showed that the expression of Caspase-3 protein in HXO-RB44 cells was gradually increased with the increase of the concentration of SalB. CONCLUSION: SalB can significantly affect on HXO-RB44 cells growth inhibition and apoptosis induction which may be achieved through the up-regulation of Caspase-3 expression and the induction of cell cycle arrest.