Synergistic protective effect of astragaloside IV-tetramethylpyrazine against cerebral ischemic-reperfusion injury induced by transient focal ischemia

Ethnopharmacological relevance

Astragaloside IV and tetramethylpyrazine have been extensively used in the cardio-cerbrovascular diseases of medicine as a chief ingredient of glycoside or alkaloid formulations for the treatment of stroke and myocardial ischemia diseases.

Aim of the study

To investigate the effects of astragaloside IV (ASG IV) and tetramethylpyrazine (TMPZ) on cerebral ischemia-reperfusion (IR) injury model in rat model.

Materials and methods

Rats were randomly divided into the following five groups: sham group, IR group and treatment group including ASG IV, ASG IV–TMPZ and nimodipine treatment. The therapeutic effect was evaluated by micro-positron emission tomography (Micro-PET) using ¹⁸F-fluoro-2-deoxy-d-glucose. The neurological examination, infarct volume and the levels of oxidative stress- and cell apoptosis-related molecules were assessed.

Results

Micro-PET imaging showed that glucose metabolism in the right hippocampus was significantly decreased in the IR group compared to the sham group (P < 0.01). ASG IV and ASG IV–TMPZ treatments reversed the decreased glucose metabolism in the model group (P < 0.05 and P < 0.01, respectively). IR induced the increase of Caspase-3 mRNA levels, MDA content and iNOS activity, but it caused the decrease of SOD activity and Bcl-2 expression compared the sham group (P < 0.01). ASG IV–TMPZ and ASG IV reversed the IR-induced changes of these parameters, i.e. the down regulation of Caspase-3 mRNA, MDA content and iNOS activity, and the up regulation of SOD activity and Bcl-2 expression (P < 0.05).

Conclusion

This study showed that ASG IV–TMPZ played a pivotal synergistic protective role against focal cerebral ischemic reperfusion damage in a rat experimental model.     

Anti-hyperuricemic and nephroprotective effects of Modified Simiao Decoction in hyperuricemic mice

Ethnopharmacological relevance

Modified Simiao Decoction (MSD), based on clinical experience, has been used for decades and famous for its efficiency in treating hyperuricemic and gouty diseases.

Aim of the study

To investigate the effects of MSD on anti-hyperuricemic and nephroprotective effects are involved in potassium oxonate-induced hyperuricemic mice.

Materials and methods

The effects of MSD were investigated in hyperuricemic mice induced by potassium oxonate. MSD were fed to hyperuricemic mice daily at a dose of 0.45, 0.90, 1.80 g/kg for 10 days, and allopurinol (5 mg/kg) was given as a positive control. Serum and urine levels of uric acid and creatinine, and fractional excretion of uric acid (FEUA) were determined by colorimetric method. Its nephroprotective effects were evaluated by determining a panel of oxidative stress markers after the intervention in hyperuricemic mice. Simultaneously, protein levels of urate transporter 1 (URAT1) and organic anion transporter 1 (OAT1) in the kidney were analyzed by Western blotting.

Results

MSD could inhibit XOD activities in serum and liver, decrease levels of serum uric acid, serum creatinine and BUN, and increased levels of urine uric acid, urine creatinine, FEUA dose-dependently through down-regulation of URAT1 and up-regulation of OAT1 protein expressions in the renal tissue of hyperuricemic mice. It also effectively reversed oxonate-induced alterations on renal MDA levels and SOD activities in this model.

Conclusion

MSD processes uricosuric and nephroprotective actions by regulating renal urate transporters and enhancing antioxidant enzymes activities to improve renal dysfunction in hyperuricemic mice.     

Antioxidative and hepatoprotective effect of CGX, an herbal medicine, against toxic acute injury in mice

Aim

CGX, a modified traditional Chinese herbal drug whose name means “liver cleaning,” is used to treat various liver disorders. This study investigated the protective effects of CGX and its mechanisms.

Material and methods

After pretreating ICR mice twice daily with CGX (po, 50 or 100 mg/kg) or distilled water for three consecutive days, acute liver injury was induced by a single injection of CCl₄ (ip, 10 mL/kg of 0.2% in olive oil) (n = 8 per group).

Results

Pretreatment with CGX significantly attenuated the elevation in biochemical parameters, such as alanine transaminase (ALT), aspartate transaminase (AST), and lactate dehydrogenase (LDH) in serum, and the malondialdehyde concentrations in liver tissue. Pretreatment with CGX significantly restored the reduction of catalase activity and glutathione (GSH) content, but not superoxide dismutase (SOD) activity, and it inhibited the CCl₄-induced high expression of iNOS and TNF-α in hepatic tissue.

Conclusion

This study showed that CGX has hepatoprotective effects against free radical-induced acute injury via primarily antioxidative properties.     

Neuroprotective activityof Cymbopogon martinii against cerebral ischemia/reperfusion-induced oxidative stress in rats

Ethnopharmacological relevance

Cymbopogon martinii (Roxb.) Watson (Family: Graminae), commonly known as Palmarosa, is traditionally prescribed for central nervous system (CNS) disorders such as neuralgia, epileptic fits and anorexia. Although the plant possesses diverse pharmacological actions, the neuroprotective action has got little attention.

Aim of the study

The present study evaluated neuroprotective effect of essential oil of Cymbopogon martinii (EOCM) against global cerebral ischemia/reperfusion (I/R)-induced oxidative stress in rats.

Materials and methods

Global ischemic brain damage was induced by bilateral common carotid artery (BCCA) occlusion for 30 min, followed by 60 min reperfusion on Wistar albino rats. The biochemical levels of lipid peroxidation (LPO), superoxide dismutase (SOD), catalase (CAT), total thiols and glutathione (GSH) were estimated and brain coronal sections and histopathological studies were performed.

Results

BCCA occlusion, followed by reperfusion caused varied biochemical/enzymatic alterations viz. increase in LPO and decrease in SOD, CAT, total thiols and GSH. The prior treatment of EOCM (50 mg/kg and 100 mg/kg, p.o. for 10 days) markedly reversed these changes and restored to normal levels as compared to I/R groups. Moreover, brain coronal sections and histopathological studies revealed protection against ischemic brain damage in the EOCM-treated groups.

Conclusion

This study, for the first time, shows potent neuroprotective effect of EOCM against global cerebral I/R-induced oxidative stress in rats, suggesting its therapeutic potential in cerebrovascular diseases (CVD) including stroke.     

Effect of essential oil of Syringa pinnatifolia Hemsl. var. alashanensis on ischemia of myocardium, hypoxia and platelet aggregation

Aim of the study

To investigate the cardioprotective potential of Syringa pinnatifolia Hems1. var. alashanensis essential oil (SPEO) against experimental acute myocardial ischemia (AMI), hydrogen peroxide (H₂O₂)-induced myocyte injury and activities against hypoxia and platelet aggregation.

Materials and methods

Wistar rats, Kunming mice and primary cultured rat neonatal myocytes were used in this study. AMI in rats was induced by ligation of the left anterior descending (LAD) coronary artery, and deviation of ST-segment, as well as changes of myocardial enzyme activities were observed. Hypoxia test in Kunming mice was performed to evaluate the effect of SPEO against hypoxia. The protective effect of SPEO on H₂O₂-induced cell injury was evaluated in terms of cell viability assay. The in vitro effect of SPEO against platelet aggregation was studied using adenosine 5′-diphosphate (ADP) as agonist.

Results

Administration of SPEO reduced deviation of ST-segment, decreased the levels of lactate dehydrogenase (LDH), creatine kinase (CK) and Troponin T (TnT) while increased the activities of superoxide dismutase (SOD). The protective role of SPEO was further confirmed by histopathological examination. In the hypoxia test, both 8 and 32 mg/kg of SPEO could prolong survival time of mice under hypoxia condition. At the meantime SPEO showed remarkable protective effect on cultured rat myocyte death induced by H₂O₂. SPEO also inhibited ADP-induced rat platelet aggregation by 47.4%, 37.0% and 32.9% at the dose of 5, 2.5 and 1.25 μg/mL, respectively.

Conclusions

These results suggest that SPEO possessing activities against hypoxia, oxidative stress and platelet aggregation has a significant protective effect against experimental myocardial ischemia.     

Extracts from Astragalus membranaceus limit myocardial cell death and improve cardiac function in a rat model of myocardial ischemia

Ethnopharmacological relevance

The root of Astragalus membranaceus, known as “huang-qi”, is one of the most widely used Chinese herbal medicines for the prevention and treatment of myocardial ischemic diseases. However, the mechanisms governing its therapeutic effects are largely unknown.

Aims of the study

The aims of the present study were to investigate the cardioprotective effect of the root extract of Astragalu membranaceus (EAM) in myocardial ischemia and to explore its underlying mechanisms in ROS-mediated signaling cascade in vivo and in vitro.

Materials and methods

The saponins in EAM were analyzed using HPLC. The tests for the cardioprotective effects of EAM and its mechanisms were performed in vivo and in vitro. In vivo, the rat model of persistent myocardial ischemia was produced by occlusion of the left anterior descending (LAD) coronary artery. In vitro, the cardiomyocyte model of oxidative stress was mimicked by the direct free radical donor, H₂O₂.

Results

In vivo, the increased myocardial infarct size and the increased serum levels of lactate dehydrogenase (LDH), creatine kinase isoform MB (CK-MB), and cardiac troponin (cTnI) were significantly decreased by pre-treatment with EAM. Moreover, cardiac function, as assessed by±dP/dt, left ventricular developed pressure (LVDP), and left ventricular end-diastolic pressure (LVEDP), was dramatically improved. An oxidative stress biomarker, malondialdehyde (MDA), was reduced, and the antioxidant enzyme superoxide dismutase (SOD) was induced. In vitro, H₂O₂-triggered myocardial cell death and cytoplasm Ca²⁺ overload were blocked by treatment with EAM. Furthermore, the K_ATP channel blocker (5-HD, glibenclamide) blocked the anti-apoptotic protective effect of EAM on cardiomyocytes injured by H₂O₂.

Conclusions

The cardioprotection of EAM was manifested as a protection of tissue structure and as a decrease in serum markers of ischemic injury. The mechanisms underlying the EAM-mediated protective effects may involve improving cardiac function, attenuating the oxidative injury via a decrease in MDA, a maintenance in SOD, and a reduction in free radical-induced myocardial cell injury. Additionally, EAM enhanced the myocardial cell viability via arresting the influx of Ca²⁺ to block cell death and opening mitochondrial K_ATP channels to reduce cell apoptosis.     

The protective effect of Zizyphus jujube fruit on carbon tetrachloride-induced hepatic injury in mice by anti-oxidative activities

Aim of the study

The study was aimed to investigate the protective effect against hepatic injury induced by CCl₄ for the ethanolic extract of FZJ.

Materials and methods

The alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected as biomarker in blood of hepatic injury. Product of lipid peroxidation (MDA), activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px), and reduced glutathione (GSH) contents were evaluated for oxidative stress in hepatic injury. Moreover, histopathological observation was assayed at the degree of hepatic injury.

Results

After administrated the FZJ, the dose of 200 mg/kg significantly decreased ALT and AST, and attenuated histopathology of hepatic injury, and ameliorated the oxidative stress in hepatic tissue. Partly assayed indexes were ameliorated after administrated FZJ at the dose of 100 mg/kg.

Conclusion

These results indicated that hepatic protective effects of FZJ were very relevant to modulate the oxidative stress in hepatic injury.     

Cardioprotective effect of water and ethanol extract of Salvia miltiorrhiza in an experimental model of myocardial infarction

Ethnopharmacological relevance

Salvia miltiorrhiza has long been used in the traditional Chinese formulations for the treatment of heart ischemic diseases.

Aim of the study

We investigated the cardioprotective effect of purified Salvia miltiorrhiza extract (SME) in an experimental model of acute myocardial infarction.

Materials and methods

Following induction of acute myocardial infarction in rats by adminstration of isoproterenol, hemodynamic and electrocardiographic parameters were monitored and recorded continuously, cardiac enzymes and parameters of oxidative stress were measured, and histopathological examination of heart tissue was performed. Experiments were performed in rats treated with SME or vehicle, as well as in those treated with Fufang Danshen Tablet (FDT) as a positive control which has previously been shown to prevent myocardial ischemia.

Results

Isoproterenol-treated rats showed reductions in left ventricular systolic pressure as well as in maximum and minimum rate of developed left ventricular pressure, together with an increase in left ventricular end-diastolic pressure. They also demonstrated ST-segment elevation, together with increases in serum levels of lactate dehydrogenase, glutamic oxalacetic transaminase, creatine kinase and malondialdehyde, as well as decreases in serum activities of glutathione peroxidase and superoxide dismutase. Oral administration of SME (29.76 or 59.52 mg/kg) blunted all of the hemodynamic and biochemical changes induced by isoproterenol, as did FDT (1210 mg/kg). The protective effect of SME on isoproterenol-induced myocardial damage was further confirmed by histopathological examination.

Conclusions

Our results suggest that SME affords protection against isoproterenol-induced myocardial infarction.     

Protective effect of Danhong injection on cerebral ischemia–reperfusion injury in rats

Ethnopharmacological relevance

Danhong injection (DH), a Chinese medical product, is used extensively for the treatment of cerebrovascular diseases such as acutely cerebral infarction in clinic.

Aim of the study

To explore the protective effect and the relevant mechanisms of DH on cerebral ischemia–reperfusion (I/R) injury.

Materials and methods

Cerebral I/R injury was induced through four-vessel occlusion (4-VO) or middle cerebral artery occlusion (MCAO). Adult male SD rats were randomly divided into six kinds of groups: normal control group, sham-operated group, I/R injury group, DH-treated groups at doses of 0.5 ml/kg, 1.0 ml/kg and 2.0 ml/kg. The effects of DH on murine neurological deficits and cerebral infarct volume, 6-keto-prostagladin F_1α (6-keto-PGF_1α) level, malondialdehyde (MDA) level and superoxide dismutase (SOD) activity in brain tissue, as well as the activities of plasma tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI) after I/R were evaluated. Moreover, the expressions of Bcl-2 and Bax protein were detected by immunohistochemistry.

Results

There was no significant difference between the control group and the sham-operated group based on the measurement indicators. Compared with the vehicle-treated group, rats treated with DH showed dose dependent reductions in brain infarction size, and improvement of neurological outcome. The level of 6-keto-PGF_1α and the activities of SOD and plasma t-PA were enhanced significantly, whereas the level of MDA and the activity of plasma PAI were declined significantly. The immunohistochemical staining results also revealed that the expression of Bcl-2 protein was up-regulated and that of Bax protein was down-regulated when exposed to DH.

Conclusion

DH demonstrates a strong ameliorative effect on cerebral I/R damage in rats by its anticoagulant, antithrombotic, antifibrinolytic and antioxidant activities. Furthermore, suppressing apoptosis through regulating Bcl-2 and Bax protein expressions should be another potential mechanism by which DH exerts its neuroprotective function.     

Protective effect of hydroalcoholic extract of Mimusops elengi Linn. flowers against middle cerebral artery occlusion induced brain injury in rats

Ethnopharmacological relevance

In the traditional Indian and Thai system of medicine, Mimusops elengi Linn., flower is used as brain tonic and to calm anxiety and panic attacks.

Aim of the study

The present study was designed to investigate the neuroprotective effect of hydroalcoholic extract of Mimusops elengi (ME) against cerebral ischemic reperfusion injury in rats.

Materials and methods

Male rats were pretreated with ME (100 and 200 mg/kg) for seven days and focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) method. After 60 min of MCAO and 24 h of reperfusion, a battery of behavioral tests assessed the extent of neurological deficits. Infarct volume and brain edema were measured in TTC stained brain sections and the extent of blood brain barrier (BBB) disruption was observed by Evan's blue extravasation. Oxidative and nitrative stress parameters were estimated in the brain homogenates. Further, simultaneous quantification of five polyphenolic biomarkers were done using HPLC.

Results

Pretreatment with ME at doses of 100 and 200 mg/kg significantly improved the neurobehavioral alterations and reduced the infarct volume, edema and extent of BBB disruption induced by ischemia reperfusion injury. It also prevented the alteration in the antioxidant status and reduced the nitrite levels when compared to ischemic animals. Further, HPLC studies revealed that ME contains five bioactive polyphenolic compounds.

Conclusions

These results clearly indicate the neuroprotective effect of ME against stroke like injury. The observed protective effect might be attributed to the polyphenolic compounds and their antioxidant and anti-inflammatory property.     

Antifibrotic effects of Artemisia capillaris and Artemisia iwayomogi in a carbon tetrachloride-induced chronic hepatic fibrosis animal model

Ethnopharmacological relevance

Artemisia capillaris and Artemisia iwayomogi, both members of the Compositae family, have been indiscriminately used for various liver disorders as traditional hepatotherapeutic medicines in Korea for many years.

Aim of the study

In this study, the anti-hepatofibrotic effects of Artemisia capillaris and Artemisia iwayomogi were comparatively analyzed using a carbon tetrachloride (CCl₄)-induced liver fibrosis rat model.

Materials and methods

Hepatic fibrosis was induced via a 10-week course of intraperitoneal CCl₄ injections (50% dissolved in olive oil, 2 mL/kg, twice per week). Water extract of Artemisia capillaris (AC) or Artemisia iwayomogi (AI) was orally administered six times per week from the 5th to the 10th week.

Results

AI (50 mg/kg) significantly attenuated the CCl₄-induced excessive release of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) in serum (p < 0.05), and hydroxyproline and malondialdehyde (MDA) contents in liver tissue (p < 0.05). Further, AI markedly ameliorated the depletion of total antioxidant capacity (TAC), glutathione (GSH), and superoxide dismutase (SOD) in liver tissue (p < 0.01). Unexpectedly, AC did not exert any effects on the above parameters. Histopathological and immunohistochemical analyses revealed that AI drastically reduced inflammation, necrosis, fatty infiltration, collagen accumulation, and activation of hepatic satellite cells in liver tissue. These changes were not observed with AC treatment. Several critical genes of fibrosis-related cytokines including transforming growth factor beta (TGF-β), platelet-derived growth factor beta (PDGF-β), and alpha smooth muscle actin (α-SMA) were more prominently downregulated by AI compared to AC treatment.

Conclusion

Our results show that AI exerts greater hepatoprotective and anti-fibrotic effects as compared with AC via enhancing antioxidant capacity and downregulating fibrogentic cytokines.     

A study on antioxidant, free radical scavenging, anti-inflammatory and hepatoprotective actions of Aegiceras corniculatum (stem) extracts

AIM OF THE STUDY: The present study was conducted to evaluate the antioxidant, anti-inflammatory and hepatoprotective potential of Aegiceras corniculatum Linn. Blanco (Aegicerataceae). METHODS AND RESULTS: The n-hexane, ethyl acetate and methanol extracts, derived from Aegiceras corniculatum stems, scavenged superoxide anions (O2*) and hydroxyl radicals (*OH) in nitro blue tetrazolium reduction and deoxyribose degradation assays, respectively. All the extracts inhibited the process of lipid peroxidation at its initiation step. Additionally, in rat liver microsomes n-hexane and ethyl acetate extracts also caused termination of radical chain reaction supporting their scavenging action towards lipid peroxy radicals (LOO*). Moreover, increased production of O2* in human neutrophils, stimulated by phorbol-12-myristate-13-acetate (PMA) and/or opsonized zymosan were also suppressed (IC50 approximately 3-20 microg/mL). Thereby, revealing the ability of plant extracts to antagonize the oxidative stress via interference with NADPH oxidase metabolic pathway. These in vitro results coincide with the reduction in the glucose oxidase-induced paw edema in mice in the presence of ethyl acetate and methanol extracts (10, 50, and 100mg/kg, i.p.). Plant extracts (250, 500 and 1000 mg/kg, p.o.) also significantly protected the carbon tetrachloride (CCl4)-induced oxidative tissue injury in rat liver. This was reflected by a approximately 60% decline in the levels of serum aminotransferase enzymes. CONCLUSION: Aegiceras corniculatum extracts found to possess pronounced antioxidant effect that may be at least in part related to its anti-inflammatory and hepatoprotective activities. This study provides a scientific basis for the ethnomedical claims that Aegiceras corniculatum is effective against inflammation and liver injury.     

Andrographolide inhibits the expression and metabolic activity of cytochrome P450 3A4 in the modified Caco-2 cells

Aim of the study

The aim of this study is to examine the effects of andrographolide on intestinal enzyme cytochrome P450 3A4 (CYP3A4) and predict whether oral administration of andrographolide-containing remedy leads to herb–drug interaction.

Materials and methods

Caco-2 cells are treated with 1α, 25-dihydroxyvitamin D3 for 3 wks to induce the expression of CYP3A4, and then andrographolide (1, 10, 100 μM) is added and treated for 72 h. Upon the further 4-h testosterone (250 μM) or nifedipine (200 μM) treatment, the basolateral medium samples and the Caco-2 monolayers are collected for analyses.

Results

Andrographolide (1, 10, 100 μM) significantly down-regulates the mRNA level and protein level of CYP3A4, and inhibits nifedipine oxidation and testosterone 6β-hydroxylation.

Conclusion

Oral administration of andrographolide likely leads to reduction of the metabolic activity of intestinal CYP3A4, therefore herb preparations containing andrographolide may result to herb–drug interactions in combination therapy.     

Role of Sida cordifolia L. leaves on biochemical and antioxidant profile during myocardial injury

Ethnopharmacological relevance

The Sida cordifolia L. (Family: Malvaceae) is a widely allocated herb by folk tribes of Gujarat state of India for the treatment of coronary manifestations. However, no published data relevant to use of the plant is available.

Aim of the study

The aim of the present study was to evaluate the antioxidant and biochemical profile of hydroalcoholic extract of Sida cordifolia L. (HESC) leaves against myocardial infarction (MI) in rats.

Materials and methods

Albino rats were administered HESC (100 and 500 mg/kg) and propranolol (10 mg/kg) once daily orally for 30 days. At the end of treatment period, MI was induced by administering isoproterenol (ISO) or by subjecting heart to ischemia reperfusion injury (IRI). Endogenous biomarkers (LDH and CK-MB) and antioxidants (SOD and catalase) were estimated in serum/perfusate and heart tissue homogenate (HTH).

Results

The LDH and CK-MB activities were elevated in HTH and depleted in serum/perfusate of HESC and propranolol groups when compared to ISO/IRI control. Further, it was found that both doses significantly increased endogenous antioxidants in HTH. Moreover, biochemical findings were supported by histopathological observations.

Conclusion

The result confirm, at least in part, for the use of Sida cordifolia in folk medicine to treat MI.     

Protective effect of Heliotropium foertherianum (Boraginaceae) folk remedy and its active compound, rosmarinic acid, against a Pacific ciguatoxin

Ethnopharmacological relevance

Terminalia pallida is an evergreen endemic tree, mentioned in Ayurveda as the fruits of Terminalia pallida are excellent in cardioprotective property. Tribal people use Terminalia pallida fruit for the treatment of diabetes and this plant widely used in many other disorders.

Aim of study

The present investigation was to evaluate the antioxidant, biochemical profile and histological studies of qualitatively standardized ethanolic extract of Terminalia pallida fruits (TpFE) against isoproterenol-induced myocardial infarction in rats.

Materials and methods

TpFE was standardized by high performance liquid chromatography (HPLC) and mass spectroscopy (MS). Rats were pretreated orally with different doses of TpFE (100, 300, and 500 mg kg⁻¹ body weight) and cardioprotective positive control gallic acid (GA) for 30 days prior to isoproterenol (ISO) induced myocardial infarction. The rats were sacrificed, hearts were collected and homogenized for biochemical analysis. The effects on total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C) and very low density lipoprotein cholesterol (VLDL-C), high density lipoprotein cholesterol (HDL-C), lipid peroxidation (LPO) marker, malondialdehyde (MDA), creatine kinase (CK), lactate dehydrogenase (LDH), alanine transaminase (ALT), aspartate transaminase (AST), catalase (CAT), glutathione peroxidase (GPx), sodium potassium (Na⁺/K⁺), calcium (Ca²⁺) and magnesium (Mg²⁺) adenosine triphosphatases (ATPases) were estimated in heart tissue homogenate.

Results

Rats administered with ISO showed a significant increase in TC, TG, LDL-C, VLDL-C, and MDA and a significant decrease in HDL-C, cardiac marker enzymes – CK, LDH, ALT and AST. ISO significantly reduced antioxidants – CAT, GPx, and membrane bound enzymes – Na⁺/K⁺, Ca²⁺ and Mg²⁺ ATPases. Pretreatment with TpFE (100, 300, and 500 mg kg⁻¹ bw) and GA (15 mg kg⁻¹ bw) for a period of 30 days significantly inhibited the effects of ISO. Moreover, biochemical findings were supported by histopathological observations.

Conclusion

The present study provide evidence for the first time, that TpFE pretreatment ameliorated myocardial injury in ISO-induced myocardial infarcted rats and exhibited cardioprotective activity.     

Proteomic studies on protective effects of salvianolic acids, notoginsengnosides and combination of salvianolic acids and notoginsengnosides against cardiac ischemic-reperfusion injury

Ethnopharmacological relevance

Salvia miltiorrhiza and Panax notoginseng are popularly used traditional Chinese medicine for cardiovascular disorders and they are often used in the form of combination. However, mechanisms of their cardioprotective effects were still not clear. In the present study, the protective effects of salvianolic acids (SA), notoginsengnosides (NG) and combination of SA and NG (CSN) against rat cardiac ischemia–reperfusion injury were checked and the protein expression profiles of heart tissues were examined to search their possible protein targets.

Materials and methods

The cardioprotective effects of SA, NG and CSN were checked in a rat model of ischemia–reperfusion (IR) by temporarily occluding coronary artery for 20 min followed by reperfusion. Rats were grouped into sham-operation group, IR group, IR + SA group, IR + NG group and IR + CSN group. The plasma creatine kinase (CK) activities were measured using commercial kit and the percentages of infarcted area in total ventricle tissue were calculated after nitroblue-tetrazolium (N-BT) staining of heart tissue slices. Two-dimensional protein electrophoresis (2-DE) was used to check the protein expression profiles of heart tissues. Then, proteins differentially expressed between IR group and sham-operation group were identified using matrix assisted laser desorption ionization-time of flight-mass spectrometry/mass spectrometry (MALDI-TOF MS/MS). The regulative effects of SA, NG and CSN on these IR-related proteins were analyzed.

Results

Treatments including SA, NG and CSN all showed cardioprotective effects against ischemia–reperfusion injury and CSN exhibited to be the best. Eighteen proteins involved in IR injury were found. These proteins are involved in pathways including energy metabolism, lipid metabolism, muscle contraction, heat shock stress, cell survival and proliferation. The regulation of these proteins by SA, NG or CSN suggested possible protein targets in their cardioprotective effects.

Conclusions

SA and NG showed both similarity and difference in their protein targets involved in cardioprotective effects. The capability of CSN to regulate both protein targets of SA and NG might be the basis of CSN to show cardioprotective effects better than that of SA or NG.