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Objective@#To investigate the effect and related mechanism of salvianolate on myocardial ischemia-reperfusion (I/R) injury in rats.@*Methods@#Thirty-six adult Sprague-Dawley rats were divided into three groups (n=12 each) using random number table method: control group (coronary artery was not ligated) , I/R group (myocardial I/R model was established by ligation and opening of left anterior descending coronary artery) , and salvianolate+I/R group (5 mg/kg of salvianolate was injected through the tail vein at the time of reperfusion) .Triphenyltetrazolium chloride (TTC) stain was utilized to measure the myocardial infarct size. The ELISA method was used to detect myocardial necrotic markers, including cardiac troponin T (TnT) , creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) . Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was used to analyses the levels of apoptosis. The levels of cleaved Caspase-3 protein were analyzed with Western blot.Cold luminescence method was used to detect the ATP level of myocardial tissue. The levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG) in myocardial tissue were detected by immunofluorescence.@*Results@#(1) The infarct size in control group, I/R group and salvianolate+I/R group were 0, (23.90±5.66) %, and (12.06±5.97) %, respectively (P<0.01 or 0.05) . (2) The TnT level was (0.04±0.03) , (16.96±2.80) , and (6.95±2.31) ng/ml, the CK-MB level was (43.6±23.5) , (1 135.8±180.6) ,and (390.3±121.5) U/L, the LDH level was (119.0±58.6) , (1 838.6±543.8) , and (631.6±228.3) U/L in control group, I/R group and salvianolate+I/R group, all significantly lower in salvianolate+I/R group than in I/R group (all P<0.01) . (3) The rate of TUNEL positive myocardial cells were (1.07±1.16) %, (21.36±4.11) %,and (13.30±3.67) % in control group, I/R group,and salvianolate+I/R group (all P<0.01) . (4) The cleaved Caspase-3 expression was 0.11±0.05, 0.84±0.20,and 0.43±0.09 in control group, I/R group, and salvianolate+I/R group (all P<0.01) . (5) The ATP level of myocardial tissue was (100.0±0.0) %, (34.2±9.2) %,and (62.1±18.0) % respectively in control group, I/R group, and salvianolate+I/R group (all P<0.01) . (6) There was almost no 8-OHdG expression in the myocardial tissue of control group. The expression of 8-OHdG in the myocardial tissue of I/R group was greater than that of the control group. The expression of 8-OHdG in the myocardial tissue of salvianolate+I/R group was less than that of the I/R group.@*Conclusion@#Salvianolate may alleviate myocardial I/R injury of rat through reducing the mitochondrial DNA oxidative damage, protecting mitochondrial function and inhibiting the apoptosis of myocardial cells.
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Objective@#To investigate the possible mechanism related to the protective effects of salvianolate in H9c2 cells underwent hypoxia/reoxygenation (H/R)-injury.@*Methods@#H9c2 cells were divided into four groups: control group, salvianolate group (S group), H/R group, and salvianolate+ H/R group(S+ H/R group), in which the H9c2 cells were pretreated with salvianolate before H/R-treatment.Apoptotic cells were detected by Tunel assays and AnnexinⅤ-FITC apoptosis detection kit.The intracellular ATP level, the change of mitochondrial membrane potential and the mitochondrial DNA oxidative damage were also determined in these groups.@*Results@#(1) The apoptosis rate of H/R group(26.36±5.14)% was significantly higher compared to control group(2.71±1.66)%(P=0.000 4), which could be significantly reduced in S+ H/R group(17.28±4.75)%(P=0.012 8 vs. H/R group , P=0.003 9 vs. control group). The ratio of AnnexinⅤ and PI double positive cells in H/R group(28.23±6.73)% was significantly higher compared to control group(3.53±2.83)%(P=0.001 1), which was significantly reduced in S+ H/R group(18.10±4.56)%(P=0.037 2 vs. H/R group, P=0.038 3 vs. control group). (2)The ATP level of H9c2 cells in H/R group(49.05±10.12)% was significantly lower than in control group 100%(P=0.000 5), which was significantly increased in S+ H/R group(68.67±13.32)%(P=0.019 9 vs. H/R group). Confocal microscope showed that red fluorescence was dominant in the control group, red fluorescence was significantly reduced, while green fluorescence was significantly increased in H9c2 cells of H/R group and the fluorescence ratio of red to green in H/R group((37.13±8.47)%) was significantly decreased compared to control group (100%, P=0.000 1), fluorescence ratio of red to green was significantly increased in S+ H/R group((63.77±12.32)% vs. H/R group, P=0.007 3). (3)The mitochondrial DNA oxidative damage in different groups: there was only few 8-hydroxyguanine (8-OHdG) expression, which marked as green, in control group, and 8-OHdG expression was significantly upregulated in H/R group, moreover, the 8-OHdG was co-localized with mitochondria.The expression of 8-OHdG was significantly lower in S+ H/R group compared to H/R group.@*Conclusion@#Salvianolate can reduce mitochondrial DNA oxidative damage, and protect mitochondrial function, thus inhibit myocardial cell apoptosis and eventually reduce the myocardial H/R-injury in H9c2 cells.
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Objective To investigate the possible mechanism of metoprolol on protecting against ischemia‐reperfusion in‐duced injury .Methods A total of 32 healthy 3-4 months male C57BL/6 mice were divided into four groups(n=8)as following :Sham‐operating group(control group);metoprolol group;ischemia‐reperfusion group(I/R group);metoprolol + I/R group .Myo‐cardial injury ,apoptosis ,cytochrome c release ,Caspase‐3 activity and calpain activity were determined in these groups .Results Al‐though there was no obvious changes in the regions at risk between I/R group and metoprolol + I/R group ,metoprolol pretreat‐ment significantly reduced the ratio of the infarct to risk regions(P<0 .05) .In the I/R group ,the rate of cardiomyocyte apoptosis , cytochrome c release ,as well as the activity of Caspase‐3 and calpain significantly increased compared to the control group(P<0 .01) .However ,these effects of I/R injury were alleviated by pretreatment with metoprolol .Conclusion metoprolol might protect against ischemia‐reperfusion induced injury by preventing calpain activation .
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Objective To observe the effect of sinomenine(SINO) on immune function of adjuvant arthritis(AA) rats.Methods SD rats were randomized into normal group,model group,SINO groups(treated with gastric gavage of SINO at the doses of 60,120 and 240 mg/kg respectively),and glucosides of Tripterygium Wilfordii(30 mg/kg) group.Except the normal group,the rats in other groups received subcutaneous injection of the complete Freund's adjuvant 0.1mL into the left hind foot to induce AA.The medication began from 12 days after the modeling and lasted 12 days.The pedal swelling and joint function scores were observed in different time.Radioimmunoassay was used to detect the contents of interleukin-1?(IL-1?) and tumor necrosis factor ?(TNF-?) in synovial cells.Expression of IL-1? and TNF-? mRNA was examined by reverse transcription-polymerase chain reaction(RT-PCR) assay.Results SINO at different concentrations decreased the pedal swelling and arthritis scores to various degrees,inhibited the production of IL-1? and TNF-? in the synovial cells,reduced the expression of IL-1? and TNF-? mRNA,and recovered the normal histological features of synovial cells in AA rats.Conclusion The therapeutic mechanism of SINO for rheumatoid arthritis may be related with the inhibition of secretion of inflammatory mediators in synovial cells,and with the recovery of histological features of synovial cells.
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Aechmea fasciata is a bromeliad that is propagated by tissue culture as an ornamental plant. A high percentage (25 to 55%) of 1- to 5-month-old seedlings were found decayed in nursery gardens that have been established in recent years in Hainan Province, People's Republic of China. There were two types of symptoms: (i) in yellow rot, the decay appears first in the new leaf apex as a water-soaked, yellow lesion, and the lesion spreads from the leaf apex to the leaf base until the whole leaf becomes water soaked and yellow; and (ii) in brown rot, decay occurs first at the base of older leaves. The lesion is water soaked but later becomes brown, and the lesions develop from leaf base to leaf apex and spread to adjacent leaves. Eventually the whole plant is rotted, but the decay remains brown. Both types of symptoms on A. fasciata have not been reported in China. Fusarium sp. was isolated from the yellow rot type lesions, and Pythium sp. was isolated from the brown rot type lesions. Isolates of Fusarium and Pythium spp. were inoculated on wounded and unwounded healthy plants. Yellow rot type and brown rot type lesions were observed on wounded host three days after inoculation with the respective pathogen. No lesions were observed on unwounded inoculated plants or on wounded and unwounded control plants. The same Fusarium and Pythium spp. were reisolated from the yellow and brown rot type lesions, respectively, in inoculated plants, thus fulfilling Koch's postulates. Colonies of the Fusarium sp. were white on potato dextrose agar, purple on rice medium, and first yellow, and then blue green in 4 days on selective carrot medium. On carnation media, microconidia were abundant, 1- to 2-celled, oval- to kidney-shaped, and 2.4 to 3.6 × 4.8 to 7.2 µm. Macroconidia were rare, sickle-shaped with attenuated apical cells and foot-shaped basal cells. Chlamydospores were also abundant, globose, formed singly or in pairs, and intercalary or on short lateral branches. This pathogen was identified as Fusarium oxysporum based on these characteristics (1,3). The Pythium sp. on potato dextrose agar was white and cottony. On selective carrot medium, hyphae were 3.6 to 7.5 µm in diameter. Sporangia had irregular swellings and were acrogenous. Oogonia were globose, colorless, acrogenous or intercalary, 14.8 to 27.6 µm in diameter. Antheridia were copulated with oogonia in its apex or lateral. Oospores were spherical, aplerotic, and 13.2 to 25.2 µm in diameter. This pathogen was identified as Pythium aphanidermatum based on these characters (2). To our knowledge, this is first report of leaf rot caused by F. oxysporum and P. aphanidermatum on A. fasciata in the People's Republic of China. References: (1) C. Booth. The Genus Fusarium. The Eastern Press, London, 1971. (2) W. Jingchao. Pages 30-32 in: Manual of Fungi Identification. The Shanghai Science Technology Press, 1979. (3) P. E. Nelson et al. Fusarium Species-An Illustrated Manual of Identification. The Pennsylvania State University Press University Park, 1976.
