A High-Performance Liquid Chromatography Method for the Simultaneous Determination of Five Index Components in Danhong Injection

The purpose of this study was to establish a high-performance liquid chromatography (HPLC) method for the simultaneous determination of sodium danshensu, protocatechuic aldehyde, rosmarinic acid, salvianolic acid B, and 4-coumaric acid in Danhong injection. The chromatographic method employed was as follows: the column was a Welch Ultimate XB-C18 column (250 mm × 4.6 mm, 10 μm), the mobile phase was a gradient elution of 0.4% formic acid aqueous solution (A) and acetonitrile (B), the detection wavelengths were 280 nm for sodium danshensu, protocatechuic aldehyde, and salvianolic acid B and 326 nm for 4-coumaric acid and rosmarinic acid, the sample volume was 10 μL, the flow rate was 1.0 mL/min, and the column temperature was 35°C. This method can realize the separation and determination of sodium danshensu, protocatechuic aldehyde, rosmarinic acid, salvianolic acid B, and 4-coumaric acid within 50 minutes. The linear relationships of the five peak areas and their concentrations are good (R2> 0.9997). The precision RSD values are all less than 1.0%. The reproducibility RSD values are all less than 1.3%. The stability RSD values are all less than 2.2%. The recovery values ranged from 92.4% to 99.4%. This method is simple, accurate, and reproducible. It can be used for the determination of sodium danshensu, protocatechuic aldehyde, rosmarinic acid, salvianolic acid B, and 4-coumaric acid in Danhong injection.

Protective effect of Danhong injection in patients with acute myocardial infarction at a high risk of no-reflow during primary percutaneous coronary intervention

Objective To observe the effect of Danhong injection(DI)in patients with acute ST-segment elevation myocardial infarction(STEMI)at a high risk of no-reflow(NR)during primary percutaneous coronary intervention(PCI).Methods Patients were placed in a DI group and control group.The DI group was given DI and the control group was given physiologic saline.The administration lasted 4 to 6 days in both groups after PCI.Cardiac magnetic resonance(CMR)was carried out during the perioperative period(7±2 days).The primary endpoint of the study was myocardial infarct size(IS)imaged on delayed-enhancement CMR.The secondary endpoint was major adverse cardiac events observed 6 months after PCI.Results In total,160 high-risk NR patients were enrolled,and 110 patients completed the CMR examination.According to postoperative CMR,the Myocardial Salvage Index and left ventricular ejection fraction were higher in the DI group(0.57±0.13 vs.0.48±0.17,P<0.01;49.3%±6.9%vs.46.2%±7.7%,P=0.03,respectively),whereas the IS was lower(19.7%±5.6%vs.22.2%±6.5%,P=0.04),compared with that in the control group.These differences were observed to be significant.After 6 months,the prevalence of major adverse cardiac events in the DI group decreased compared with that in the control group,but the differences were not observed to be significant(P>0.05).Conclusion The application of DI can reduce the myocardial infarct size in STEMI patients at a high risk of NR during primary PCI.

Pretreatment with Danhong injection protects the brain against ischemia-reperfusion injury

Danhong injection （DHI）, a Chinese Materia Medica standardized product extracted from Radix Salviae miltiorrhizae and Flos Carthami tinctorii, is widely used in China for treating acute isch-emic stroke. In the present study, we explored the neuroprotective efficacy of DHI in a rat model of temporary middle cerebral artery ocdusion, and evaluated the potential mechanisms under-lying its effects. Pretreatment with DHI （0.9 and 1.8 mL/kg） resulted in a significantly smaller infarct volume and better neurological scores than pretreatment with saline. Furthermore, DHI significantly reduced the permeability of the blood-brain barrier, increased occludin protein expression and decreased neutrophil infiltration, as well as profoundly suppressing the upreg-ulation of matrix metallopeptidase-9 expression seen in rats that had received vehicle. Matrix metallopeptidase-2 expression was not affected by ischemia or DHI. Moreover, DHI （1.8 mL/kg） administered 3 hours after the onset of ischemia also improved neurological scores and reduced infarct size. Our results indicate that the neuroprotective efficacy of DHI in a rat model of cerebral ischemia-reperfusion injury is mediated by a protective effect on the blood-brain barrier and the reversal of neutrophil infiltration.

Preventive effect of danhong huayu koufuye on diabetic retinopathy in rats

AIM: To study the effects of danhong huayu koufuye (DHK) on fasting blood glucose (FBG) and diabetic retinopathy (DR) in streptozotocin (517)-induced type 1 diabetic rats to facilitate the rational usage of this drug. METHODS: Diabetic rats were induced by injection of a single dose of STZ intraperitoneally at 50mg/kg. Flash electroretinogram (FERG) and oscillatory potentials (OPs) were used to measure retinal function. The microvascular perfusion of ears was performed to study the microcirculation in rats. FBG, body-weight, and 24-h urine volume, water intake and diet intake were also assessed. RESULTS: DHK had no effect on FBG in normal rats. However, STZ + DHK group were significantly different from those of Model and moved toward those of normal control. It reversed the increase in diet intake (P <= 0.05 vs model control) and the loss in body-weight (P <= 0.05 vs model control) in diabetic rats. DHK decreased the FBG of diabetic rats by 25.6% (P <= 0.05) and 37.9% P <= 0.01) after 14 and 21 days administration as compared with the model control, respectively. Moreover, DHK significantly increased the FERG b-wave amplitude by 80% (P <= 0.05 vs model control) and decreased the FERG b-wave latency by 15.3% (P <= 0.01 vs model control) after 24 days administration. The OP(1) and OP(2) amplitudes in DHK group were 2.6 (P <= 0.01) and 2.0 (P <= 0.01) times of model group after 24 days of DHK treatment, respectively. At the same time, OP(1) and OP(2) latencies in DHK group reduced by 16.0% (P <= 0.001) and 14.7% (P <= 0.001) as compared with the model control, respectively. Furthermore, the microvascular perfusion of DHK group was 2.4 times of model group (P <= 0.001) after 21 days administration. CONCLUSION: DHK had no effect on normal FBG. But it had antihyperglycemic activity, and had a preventive and therapeutic effect on DR in diabetic rats.

Danhong injection A modulator for Golgi structural stability after cerebral ischemiareperfusion injury

The cerebral ischemia-reperfusion model was established using the suture occlusion method, and rats were intraperitoneally given 8 mL/kg Danhong injection once a day prior to model establishment Rat brain tissues were harvested at 6, 24, 48, 72 hours after reperfusion. Immunohistochemical staining showed that transforming growth factor-J31 expression increased, while Golgi matrix protein GM130 expression decreased after cerebral ischemia-reperfusion. Danhong injection was shown to significantly up-regulate the expression of transforming growth factor-131 and GM130, and expres- sion levels peaked at 7 days after reperfusion. At 7 days after cerebral ischemia-reperfusion, Golgi morphology was damaged in untreated rats, while Golgi morphology breakage was not observed after intervention with Danhong injection. These experimental findings indicate that Danhong injec- tion can up-regulate the expression of transforming growth factor-131 and GM130, and maintain Golgi stability, thus playing a neuroprotective role in rats after cerebral ischemia-reperfusion.

Danhong Huayu Koufuye combined with metformin attenuated diabetic retinopathy in Zucker diabetic fatty rats

AIM: To evaluate effects of Danhong Huayu Koufuye (DHK, a Chinese medicinal formulae) alone or combined with metformin on diabetic retinopathy (DR) in Zucker diabetic fatty (ZDF) rats, an animal model of obese type -2 diabetes, and then to investigate the mechanisms. METHODS: ZDF (fa/fa) rats were administered with vehicle (distilled water), metformin, DHK, and DHK plus metformin. Electrophysiological and histological analysis were applied to evaluated effects of DHK alone or combined with metformin on DR. The levels of fasting blood glucose (FBG) and glycosylated hemoglobin (HbA1c) in blood were measured to evaluate the antihyperglycemic activity of DHK. Furthermore, levels of nitric oxide (NO), malondialdehyde (MDA) and activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) in serum were measured to study effects of DHK on oxidative stress in ZDF rats. In addition, body weight, lipidic indexes and insulin level were also assessed. RESULTS: DHK combined with metformin significantly reversed the prolongation of latency times of flash electroretinogram (FERG) and oscillatory potentials (OPs) in diabetic rats. Furthermore, DHK alone or combined with metformin showed a remarkable suppression of retinal neovascularization and amelioration of retinal internal limiting membrane morphology. Moreover, DHK alone or plus metformin reduced FBG (P<0.05), HbA1c 1094 (P<0.01) and MDA (P<0.01) levels in diabetic rats. In addition, reductions in levels of triglycerides (TG) (P<0.01) and low density lipoprotein cholesterol (LDL-c) (P<0.01 and P<0.05, respectively) were also observed in diabetic rats treated with DHK alone or plus metformin. CONCLUSION: DHK in combination with metformin had a preventive and therapeutic effect on DR in type-2 diabetic rats, and the possible mechanisms may be alleviating hyperglycemia, reducing oxidative stress and improving lipid metabolism.

Effects of adjuvant danhong injection therapy on nerve injury and platelet activation markers in patients with acute cerebral infarction

Objective:To investigate the effects of adjuvant danhong injection therapy on nerve injury and platelet activation markers in patients with acute cerebral infarction.Methods: A total of 102 patients with acute cerebral infarction who were treated in our hospital between January 2016 and September 2017 were reviewed and divided into the routine group (n=54) who received conventional therapy and the danhong injection group who received adjuvant danhong injection therapy. The differences in the contents of nerve injury indexes in serum and platelet activation markers in peripheral blood were compared between the two groups before treatment, after 3 d of treatment and after 7 d of treatment.Results: There was no statistically significant difference in the contents of nerve injury indexes in serum and platelet activation markers in peripheral blood between the two groups before treatment. After 3 d of treatment and after 7 d of treatment, copeptin, H-FABP and NSE contents in serum of danhong injection group were lower than those of routine group whereas BDNF and bFGF contents were higher than those of routine group;CD62p, CD42b, PAC-1 and PMA contents in peripheral blood were lower than those of routine group.Conclusion: Conventional therapy combined with adjuvant danhong injection therapy can effectively reduce the degree of nerve injury and inhibit the platelet activation in patients with acute cerebral infarction.

A Meta-Analysis of Danhong Injection in the Treatment of Vascular Dementia

Objectives: To evaluate the efficacy of Danhong injection in the treatment of vascular dementia. Methods: CNKI, VIP, CBM, MEDLINE, PubMed, and SCI Expanded databases were searched, and the clinical research literature of Danhong injection in the treatment of vascular dementia was collected. Literature screening and literature quality assessment were performed independently by two investigators based on inclusion criteria and the Cochrane System Evaluator’s Handbook version 5.0.0. Meta-analysis was performed using RevMan 5.0 software. Results: A total of 16 studies were included in a total of 1383 patients. Meta-analysis showed that Danhong injection combined with conventional chemotherapy can reduce the incidence of vascular dementia compared with conventional chemotherapy alone (odds ratio 3.13, 95% CI2.02 to 4.85, P < 0.0001), improved Mini-mental State Examination (MMSE)) score (SDD 1.76, 95% CI 0.67 to 2.85, P = 0.002). Conclusion: Danhong transfusion combined with chemical drugs for the treatment of vascular dementia is superior to the simple treatment of chemical drugs, which can improve the efficacy and quality of life of patients, indicating that Danhong injection has an auxiliary effect on the treatment of vascular dementia.

Effect of Danhong Injection Combined with Magnesium Polarization Solution and Nitrate in Treating Coronary Heart Disease

The objective of this study was to observe the effect of Danhong injection combined with magnesium polarized solution and nitrate on coronary heart disease.A total of 58 patients who had been diagnosed with coronary heart disease who participated in the voluntary study were selected and divided into observation group and control group.The number of patients in the two groups was 29,and the control group was mainly injected with nitroglycerin.The observation group was mainly treated with Danhong injection combined with magnesium polarized solution and nitrate ester.The main comparisons were the difference of clinical treatment effect,total cholesterol before and after treatment between the two groups of patients under different therapeutic drugs,and the differences in endothelin were recorded.The observation group was superior to the control group in terms of clinical treatment effect and total cholesterol and endothelin after treatment.Danhong injection combined with magnesium polarized solution and nitrate ester has a good effect in the treatment of coronary heart disease,which can be promoted and developed.

Clinical study on Danhong + low-dose glucocorticoid treatment of severe pneumonia combined with myocardial injury

Objective:To investigate the clinical effect of Danhong + low-dose glucocorticoid in the treatment of severe pneumonia combined with myocardial injury.Methods: 88 severe pneumonia patients complicated with myocardial injury admitted to our hospital between December 2016 and December 2018 were divided into the control group (n=44) and the study group (n=44) by random number table method. Control group received conventional western medicine + low-dose glucocorticoid treatment, study group were treated with Danhong injection on the basis of the treatment for control group, and the treatment lasted for 2 weeks. The differences in peripheral blood levels of blood gas indexes [PH value, oxygen partial pressure (PaO2) and CO2 partial pressure (PaCO2)] as well as serum contents of inflammatory mediators [interleukin-1 (IL-1), interleukin-17 (IL-17), C-reactive protein (CRP), tumor necrosis factorα (TNF-α) and monocyte chemoattractant protein 1 (MCP-1)], oxidative stress indexes [advanced oxidation protein product (AOPP), lipid hydroperoxide (LHP), total antioxidant capacity (T-AOC) and superoxide dismutase (SOD)] and myocardial injury markers [lactate dehydrogenase (LD), troponin I (TnI) and myoglobin (Mb)] were compared between the two groups of patients before treatment, after 1 week of treatment and after 2 weeks of treatment.Results: Before treatment, there were no statistically significant differences in peripheral blood levels of blood gas indexes as well as serum contents of inflammatory mediators, oxidative stress indexes and myocardial injury markers between the two groups (P>0.05). After 1 week and 2 weeks of treatment, peripheral blood PH value and PaO2 levels in the study group were higher than those in the control group, while PaCO2 levels were lower than those in the control group;IL-1, IL-17, CRP, TNF-α and MCP-1 contents were lower than those in the control group;AOPP and LHP contents were lower than those in the control group, while T-AOC and SOD contents were higher than those in the control group;LD, TnI and Mb contents were lower than those in the control group (P<0.05).Conclusion:Danhong + low-dose glucocorticoid therapy can effectively reduce the inflammatory stress and myocardial injury in severe pneumonia patients complicated with myocardial injury.

Efficacy and safety of Danhong injection in the treatment of arteriosclerosis obliterans:a meta-analysis of randomized controlled trials

Aims:To systematically evaluate the clinical efficacy and safety of danhong injection in the treatment of arteriosclerotic occlusion.Methods:Computer retrieval China knowledge network(CNKI),and Wiper Chinese journal database(VIP),and Wanfang data knowledge service platform,the Chinese biomedical literature database(CBM),Pubmed,science citation index(SCI)database,the Cochrane Library,including eight Chinese English database,to conform to the criteria for the two researchers use the recommended bias risk assessment method to evaluate the Cochrane bias risk,extracted data,and apply the Review Manager software Meta analysis and system evaluation.Results:Finally,13 studies were included,with a total number of 884 cases.The results showed that the clinical efficacy of danhong injection in the treatment of arteriosclerotic occlusion was higher than that of conventional treatment,and the difference was statistically significant[OR=3.08,95%CI(2.07,4.60),P<0.00001].The improvement of arteriosclerosis was more obvious[OR=0.15,95%CI(0.12,0.19),P<0.00001].The improvement of hemodynamics was more obvious[OR=-0.19,95%CI(-0.33,-0.04),P=0.01].No adverse reactions occurred.Conclusions:Danhong injection has higher clinical efficacy in the treatment of atherosclerotic occlusion,more obvious improvement in arteriosclerosis degree and hemodynamics,and higher safety.However,due to the low quality of literature research,more high-quality,large samples and randomized controlled double-blind clinical trials are needed to provide high-level evidence to further verify the efficacy and safety of danhong injection in the treatment of atherosclerotic occlusion.

Effect of Danhong injection combined with irbesartan on oxidative stress and renal function damage in patients with diabetic nephropathy

Objective: To study the effect of Danhong injection combined with irbesartan on oxidative stress and renal function damage in patients with diabetic nephropathy. Methods: Patients with diabetic nephropathy who were treated in hospital of Minzu University of China between April 2013 and February 2017 were selected, and random number table was used to divide them into Danhong + ARB group and ARB group who received routine hypoglycemic + Danhong injection + irbesartan therapy and routine hypoglycemic + irbesartan therapy respectively. The serum levels of renal function biochemical indexes, platelet activation indexes and oxidative stress indexes of the two groups of patients were detected before and 6 weeks after treatment. Results: After treatment, serum BUN, Cr, β2-MG, Cys-C, PECAM-1, CD62p, PAC-1, MDA, 8-OHdG and 8-iso-PGF2a levels of both groups of patients were significantly lower than those before treatment while T-AOC, Cu-Zn-SOD, Mn-SOD and GPx-1 levels were significantly higher than those before treatment and serum BUN, Cr, β2-MG, Cys-C, PECAM-1, CD62p, PAC-1, MDA, 8-OHdG and 8-iso-PGF2a levels of Danhong + ARB group after treatment were significantly lower than those of ARB group while T-AOC, Cu-Zn-SOD, Mn-SOD and GPx-1 levels were significantly higher than those of ARB group. Conclusion: Danhong injection combined with irbesartan therapy for diabetic nephropathy can reduce renal function damage and improve oxidative stress response.

Effect of Danhong injection on neuroethology changes, cell injury and apoptosis in rats with cerebral infarction

Objective:To study the effect of Danhong injection on neuroethology changes, cell injury and apoptosis in rats with cerebral infarction.Methods: SD rats were selected as experimental animals and divided into control group, cerebral infarction group and Danhong group, control group received sham operation, cerebral infarction group were made into cerebral infarction animal models and Danhong group were made into cerebral infarction animal models and then received Danhong injection for intervention. After the intervention, water maze experiment was used to assess the neuroethology changes of rats, and the serum and brain tissue were collected to determine nerve injury molecules and apoptosis molecules.Results: 2 weeks, 4 weeks and 6 weeks after intervention, the escape latency and the course to find platform of cerebral infarction group were higher than those of control group and the number of platform crossing were lower than those of control group;the escape latency and the course to find platform of Danhong group were lower than those of cerebral infarction group, and the number of platform crossing were higher than those of cerebral infarction group;6 weeks after intervention, CK-BB, LDH, NSE and S100β levels in serum as well as ROS, MDA, JNK1, JNK2, c-Jun, ATF-2 and Bax levels in brain tissue of cerebral infarction group were significantly higher than those of control group while GSH-PX, T-SOD and CAT levels in brain tissue were significantly lower than those of control group;CK-BB, LDH, NSE and S100β levels in serum as well as ROS, MDA, JNK1, JNK2, c-Jun, ATF-2 and Bax levels in brain tissue of Danhong group were significantly lower than those of cerebral infarction group while GSH-PX, T-SOD and CAT levels in brain tissue were significantly higher than those of cerebral infarction group. Conclusion:Danhong injection can reduce the neurocyte damage and inhibit the neurocyte apoptosis to improve the neurological function of rats with cerebral infarction.

Effect of Danhong injection on neuroendocrine molecules and ventricular remodeling in patients with ischemic cardiomyopathy heart failure

Objective:To study the effect of Danhong injection on neuroendocrine molecules and ventricular remodeling in patients with ischemic cardiomyopathy heart failure.Methods: 80 cases of patients with ischemic cardiomyopathy heart failure who were treated in our hospital between March 2014 and December 2016 were collected and divided into control group (n=40) and observation group (n=40) according to random number table. The control group received regular treatment, the observation group received regular + Danhong injection treatment, and both therapies lasted for 14 d a course, two sessions. Serum contents of neuroendocrine molecules and ventricular remodeling-related indexes, Doppler ultrasonic ventricular remodeling parameter levels, and so on were compared between two groups of patients before and after treatment.Results: Before treatment, serum contents of neuroendocrine molecules and ventricular remodeling-related indexes as well as Doppler ultrasonic ventricular remodeling parameter levels were not statistically different between two groups of patients. After treatment, serum neuroendocrine molecules NT-proBNP, ANP, AngⅡ, ALD and NE contents of observation group were lower than those of control group, serum ventricular remodeling indexes TGF-β1, NF-κB, CysC and FGF23 contents were lower than those of control group, and ultrasonic ventricular remodeling parameters LVMI, LVRI, RVEDV and RVESV levels were lower than those of control group.Conclusion: Adjuvant Danhong injection therapy helps optimize the neuroendocrine molecule contents and inhibit the ventricular remodeling process in patients with ischemic cardiomyopathy heart failure.

Effect of danhong injection combined with sodium ozagrel on serum IL-8, TNF-α, VEGF, TBIL and NSE in old patients with acute cerebral infarction

Objective:To study the effect of danhong injection combined with sodium ozagrel on serum IL-8, TNF-α, VEGF, TBIL and NSE in old patients with acute cerebral infarction.Methods:A total of 100 old patients with acute cerebral infarction in our hospital from August 2014 to December 2016 were enrolled in this study. The subjects were divided into the control group (n=50) and the treatment group (n=50) randomly. The control group was treated with sodium ozagrel, the treatment group was treated with danhong injection combined with sodium ozagrel, and both the two groups were treated for 14 days. The serum IL-8, TNF-α, VEGF, TBIL and NSE of the two groups before and after treatment were compared.Results:There were no significantly differences of the serum IL-8, TNF-α, VEGF, TBIL and NSE of the two groups before treatment. The serum IL-8, TNF-α, NSE levels of the two groups after treatment were significantly lower than before treatment, the serum VEGF, TBIL levels of the two groups after treatment were significantly higher than before treatment, and that of the treatment group after treatment were significantly better than the control group.Conclusion:Danhong injection combined with sodium ozagrel can significantly reduce the serum IL-8, TNF-α, NSE levels, improve the serum VEGF, TBIL levels, and reduce inflammation, promote the nerve functional recovery of the old patients with acute cerebral infarction, and it was worthy clinical application.

Effect of danhong injection combined with nadroparin calcium on serum CyS-C, CRP, IL-6, vWF, ET and VEGF in patients with acute coronary syndrome

Objective: To study the effect of danhong injection combined with nadroparin calcium on serum CyS-C, CRP, IL-6, vWF, ET and VEGF in patients with acute coronary syndrome. Methods: A total of 80 patients with acute coronary syndrome in our hospital from July 2014 to July 2017 were enrolled in this study. The subjects were divided into the control group (n=40) and the treatment group (n=40) randomly. The control group was treated with xuesaitong injection combined with nadroparin calcium;the treatment group was treated with danhong injection combined with nadroparin calcium. The two groups were treated for 2 weeks. The serum CyS-C, CRP, IL-6, vWF, ET and VEGF levels of the two groups before and after treatment were compared. Results: There were no significantly differences of the serum CyS-C, CRP, IL-6, vWF, ET and VEGF levels of the two groups before treatment. The serum CyS-C, CRP, IL-6, vWF, ET and VEGF levels of the two groups after treatment were significantly lower than before treatment, and that of the treatment group were significantly better than that of the control group. Conclusion: Danhong injection combined with nadroparin calcium can significantly reduce the serum CyS-C, CRP, IL-6, vWF, ET and VEGF levels of patients with acute coronary syndrome, and it was worthy clinical application.

Progresses in the research of Danhong Injection against the cardial ischemia-referfusion injury

Researches in the field of the myocardial ischemia-reperfusion injury are attracting the attentions of clinicians for the treatments that protect cardiac muscle cells from being injured can not only help the patients get recovery but also keep them in health. By clearing the free radicals and reducing calcium overload of myocardial cell, treatments with Danhong Injection will help myocardial cells survive from inflammatory reactions which are triggered by ischemia reperfusion so as that endothelial function will be improved and myocardial cell apoptosis will be inhibited. In all, Danhong Injection is an ideal medicine for protecting myocardial cell against ischemia reperfusion injury.

Danhong Injection Up-Regulates miR-125b in Endothelial Exosomes and Attenuates Apoptosis in Post-Infarction Myocardium

Objective:To investigate the involvement of endothelial cells(ECs)-derived exosomes in the antiapoptotic effect of Danhong Injection(DHI)and the mechanism of DHI-induced exosomal protection against postinfarction myocardial apoptosis.Methods:A mouse permanent myocardial infarction(MI)model was established,followed by a 14-day daily treatment with DHI,DHI plus GW4869(an exosomal inhibitor),or saline.Phosphatebuffered saline(PBS)-induced ECs-derived exosomes were isolated,analyzed by miRNA microarray and validated by droplet digital polymerase chain reaction(ddP CR).The exosomes induced by DHI(DHI-exo),PBS(PBS-exo),or DHI+GW4869(GW-exo)were isolated and injected into the peri-infarct zone following MI.The protective effects of DHI and DHI-exo on MI hearts were measured by echocardiography,Masson's trichrome staining,and TUNEL apoptosis assay.The Western blotting and quantitative reverse transcription PCR(qR T-PCR)were used to evaluate the expression levels of miR-125b/p53-mediated pathway components,including miR-125b,p53,Bak,Bax,and caspase-3 activities.Results:DHI significantly improved cardiac function and reduced infarct size in MI mice(P<0.01),which was abolished by the GW4869 intervention.DHI promoted the exosomal secretion in ECs(P<0.01).According to the results of exosomal miRNA microarray assay,30 differentially expressed miRNAs in the DHI-exo were identified(28 up-regulated miRNAs and 2 down-regulated miRNAs).Among them,DHI significantly elevated miR-125b level in DHI-exo and DHI-treated ECs,a recognized apoptotic inhibitor impeding p53 signaling(P<0.05).Remarkably,treatment with DHI and DHI-exo attenuated apoptosis,elevated miR-125b expression level,inhibited capsase-3 activity,and down-regulated the expression levels of proapoptotic effectors(p53,Bak,and Bax)in post-MI hearts,whereas these effects were blocked by GW4869(P<0.05 or P<0.01).Conclusion:DHI and DHI-induced exosomes inhibited apoptosis,promoted the miR-125b expression level,and regulated the p53 apoptotic pathway in post-infarction myocardium.

‘44号’抗虫黑杨基于根段组培再生和遗传转化体系的建立

[目的]以美洲黑杨和转BtCry1Ac欧洲黑杨的杂交子代‘44号’抗虫黑杨作为试验材料,采用无菌苗根基部为外植体,建立高效稳定的组培再生体系和黑杨遗传转化体系。[方法]以‘44号’抗虫黑杨无菌苗0.5~1.0 cm的根基部为试验材料,研究不同激素配比对根系不定芽的诱导和生根的影响,探索根系对草铵膦的临界耐受浓度,以农杆菌介导法进行遗传转化,并通过β-葡萄糖苷酸酶(GUS)基因染色实时监控和鉴定阳性植株。[结果]筛选出‘44号’抗虫黑杨根系诱导不定芽最优培养基为:WPM+20 g·L^(-1)蔗糖+7.8 g·L^(-1)琼脂+0.25 mg·L^(-1)6-BA+0.05 mg·L^(-1) NAA,分化率为86.67%;不定芽生根的最优培养基为:WPM+0.075 mg·L^(-1) NAA+7 g·L^(-1)琼脂+0.1 g·L^(-1) AC,生根率为75.00%。通过根系草铵膦抗性筛选梯度试验,确定根系对草铵膦筛选最适遗传转化筛选浓度为0.8 mg·L^(-1)。采用农杆菌介导法将外源基因转入到根中,经不定芽诱导和生根两个阶段获得再生植株苗,经特异引物进行PCR鉴定,31个外植体中阳性植株为9株,转化效率为29.0%。[结论]建立了‘44号’抗虫黑杨根系的再生和遗传转化体系,为黑杨派杨树的高效基因转化提供了途径的同时,也为以‘44号’抗虫黑杨为基础,通过转基因技术聚合更多的优良性状,提供了重要的技术支撑。

丹红注射液通过降低氧化应激损伤发挥抗肾纤维化的作用机制研究

目的探索丹红注射液对慢性肾衰竭大鼠肾损伤及肾纤维化的保护作用及其机制。方法采用腺嘌呤法建立大鼠慢性肾衰竭体内模型,采用叔丁基过氧化氢诱导NRK-52E细胞,建立肾损伤体外模型,HE染色评估丹红注射液对慢性肾衰竭大鼠的肾脏病理学损伤,免疫组化法检测肾损伤及纤维化的关键蛋白表达,免疫荧光法检测NRK-52E细胞GRP78蛋白表达,Western blot法进行组织和细胞中的关键蛋白定量,活性氧(ROS)荧光探针检测NRK-52E细胞中的ROS水平,JC-1检测试剂盒检测NRK-52E细胞中线粒体膜电位(MMP)水平。结果丹红注射液可以显著缓解慢性肾衰竭的肾间质纤维化状态,减轻病理损伤;抑制KIM-1、GRP78和CHOP表达(P<0.01),从而缓解内质网过度应激状态;降低纤维化胶原CollagenⅠ沉积(P<0.01),减少α-平滑肌肌动蛋白(α-SMA)(P<0.01),从而抑制肾间质纤维化形成;降低NRK-52E细胞内ROS的水平(P<0.01),减轻NRK-52E线粒体氧化损伤,稳定MMP水平。结论丹红注射液能够通过降低氧化损伤和缓解内质网过度应激状态,从而发挥抗肾纤维化的作用。