1.Differentially expressed genes of HepG2 cells treated with gecko polypeptide mixture
Yi-Meng DUAN ; Jian-Gang WANG ; Ying JIN ; Meng-Li GUO ; Leng-Xin DUAN
Chinese Journal of Pharmacology and Toxicology 2018;32(4):278-278
OBEJECTIVE Gecko has been clinically used in China for many years. It has been proved that the gecko polypeptide mixture(GPM)extracted from gecko could inhibit the growth of multiple types of tumor cells.In order to investigate the possible anti-tumor molecular mechanisms of GPM,we used RNA-seq technology to identify the differentially expressed genes of human hepatocellular carci-noma(HCC)HepG2 cells treated with or without GPM.METHODS The HepG2 cells were treated with different concentration of GPM(0,0.1,0.2,0.3,0.4 mg·mL-1)for 6 h,12 h and 24 h,respectively.MTT assay was used to detect the viability of HepG2 cells. DAPI fluorescence staining was performed to observe nucleus morphological changes of HepG2 cells.Western blot analysis was applied to observe the expres-sion of apoptosis- related proteins and endoplasmic reticulum stress (ERs)-related proteins in HepG2 cells.Flow cytometry was also applied to detect reactive oxygen species(ROS)generation.In this report, we showed that GPM could induce HepG2 cells apoptosis and influence HepG2 cells proliferation in a dose-dependent manner.We applied many analysis methods,including differentially expressed genes analysis,Gene Ontology(GO)enrichment analysis,KEGG pathway enrichment analysis,protein-protein interaction network analysis to screen out possible molecular mechanisms.RESULTS ER-nucleus signaling pathway, cellular response to stress and apoptotic processes were identified the potential anti-cancer molecular biological process of GPM.GPM may also induce apoptosis in HepG2 cells via endoplasmic reticulum stress pathway. The GPM could induce ROS generation and up-regulate ERs-related proteins. CONCLUSION The present study revealed the potential anti-tumor mechanism of GPM.
2.Measles pathogenic surveillance from 2005 to 2007 in Guangdong Province.
Leng LIU ; Huan-ying ZHENG ; Xue GUO ; Jian-qiong ZHU ; Yi-xin JI ; Wen-bo XU
Chinese Journal of Experimental and Clinical Virology 2008;22(6):406-408
OBJECTIVETo develop pathogenic surveillance on measles and to effectively isolate measles virus. To know the genetic characterizations and molecular epidemiology of wildtype measles viruses from 2005 to 2007 in Guangdong Province, and provide the scientific basis for measles control and eradication.
METHODSVero/Slam cell line were used, measles viruses were isolated from throat swabs or urine specimens collected from uspected measles patients in outbreaks and sporadic patients. A 450 nucleotides fragment of the C-terminal of the nucleoprotein (N) gene was amplified and by RT-PCR and subjected to sequence and phylogenetic analysis using Bio-Edit software.
RESULTS82 wild-type measels virus were obtained from 377 throat swabs and urine specimens from 2005 to 2007 in Guangdong Province measles lab. The measles isolation rate was 23.58% in 2005, 17.11% in 2006, 39.13% in 2007. The succeed rate of virus isolation is related to the quality of specimens collected and the days after rash occurrence.
CONCLUSIONSWe have grasped the technicalability of measles virus isolation and confirm action, and got higher isolation ratio. The wild-type measles virus isolated from Guangdong Province is of H1 genotype from 2005 to 2007, which is the same as the dominant genotype circulation.
China ; epidemiology ; Genotype ; Humans ; Measles ; diagnosis ; epidemiology ; genetics ; Measles virus ; classification ; genetics ; isolation & purification ; Molecular Epidemiology ; Molecular Sequence Data ; Phylogeny ; Plasma ; virology ; Population Surveillance ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; Urine ; virology
3.Causal Relations between Exposome and Stroke: A Mendelian Randomization Study
Hong-Qi LI ; Yi-Wei FENG ; Yu-Xiang YANG ; Xin-Yi LENG ; Prof Can ZHANG ; Shi-Dong CHEN ; Kevin KUO ; Shu-Yi HUANG ; Xue-Qing ZHANG ; Yi DONG ; Xiang HAN ; Xin CHENG ; Mei CUI ; Lan TAN ; Qiang DONG ; Jin-Tai YU
Journal of Stroke 2022;24(2):236-244
Background:
and Purpose To explore the causal relationships of elements of the exposome with ischemic stroke and its subtypes at the omics level and to provide evidence for stroke prevention. Methods We conducted a Mendelian randomization study between exposure and any ischemic stroke (AIS) and its subtypes (large-artery atherosclerotic disease [LAD], cardioembolic stroke [CE], and small vessel disease [SVD]). The exposure dataset was the UK Biobank involving 361,194 subjects, and the outcome dataset was the MEGASTROKE consortium including 52,000 participants.
Results:
We found that higher blood pressure (BP) (systolic BP: odds ratio [OR], 1.02; 95% confidence interval [CI], 1.01 to 1.04; diastolic BP: OR, 1.03; 95% CI, 1.01 to 1.05; pulse pressure: OR, 1.03; 95% CI, 1.00 to 1.06), atrial fibrillation (OR, 1.18; 95% CI, 1.13 to 1.25), and diabetes (OR, 1.13; 95% CI, 1.07 to 1.18) were significantly associated with ischemic stroke. Importantly, higher education (OR, 0.69; 95% CI, 0.60 to 0.79) decreased the risk of ischemic stroke. Higher systolic BP (OR, 1.06; 95% CI, 1.02 to 1.10), pulse pressure (OR, 1.08; 95% CI, 1.02 to 1.14), diabetes (OR, 1.28; 95% CI, 1.13 to 1.45), and coronary artery disease (OR, 1.58; 95% CI, 1.25 to 2.00) could cause LAD. Atrial fibrillation could cause CE (OR, 1.90; 95% CI, 1.71 to 2.11). For SVD, higher systolic BP (OR, 1.04; 95% CI, 1.00 to 1.07), diastolic BP (OR, 1.06; 95% CI, 1.01 to 1.12), and diabetes (OR, 1.22; 95% CI, 1.10 to 1.36) were causal factors.
Conclusions
The study revealed elements of the exposome causally linked to ischemic stroke and its subtypes, including conventional causal risk factors and novel protective factors such as higher education.
4.Effects of growth hormone supplementation on erectile function and expression of nNOS in aging rats.
Xian-Sheng ZHANG ; Yi-Xin WANG ; Yin-Fa HAN ; Zheng LI ; Zu-Qiong XIANG ; Jing LENG ; Xu-Yuan HUANG
National Journal of Andrology 2005;11(5):339-342
OBJECTIVETo explore the effects of growth hormone( GH) supplementation on erectile function and expression of nNOS in the intracavernosal nerves in aging rats.
METHODSTwenty male Sprague-Dawley rats aged 18 months were randomly divided into Groups A and B, and ten 2-month-old male Sprague-Dawley rats included in Group C. 1 U/(kg x d) GH was given to Group A, and the same volume of saline to Groups B and C. After 8 weeks of treatment, evaluation was made of the erections induced by apomorphine (APO), maximal intracavernous pressure (ICP) induced by intracavernous papaverine injection and expression of nNOS in the intracavernosal nerves by streptavidin-peroxidase immunohistochemical techniques.
RESULTSAfter 8 weeks of treatment, the erection frequency, maximal ICP and expression of nNOS in the intracavernosal nerves of the rats in Groups A and C were significantly higher than those in Group B (P < 0.05 or P < 0.01).
CONCLUSIONGrowth hormone supplementation can improve the erectile function of aging rats, which may be attributed to the increase in the expression of nNOS in the intracavernosal nerves.
Animals ; Apomorphine ; pharmacology ; Growth Hormone ; pharmacology ; Immunohistochemistry ; Male ; Nitric Oxide Synthase Type I ; biosynthesis ; Papaverine ; pharmacology ; Penile Erection ; drug effects ; Penis ; enzymology ; innervation ; Random Allocation ; Rats ; Rats, Sprague-Dawley
5.Comparison between sildenafil plus sertraline and sertraline alone in the treatment of premature ejaculation.
Xian-sheng ZHANG ; Yi-xin WANG ; Xu-yuan HUANG ; Jing LENG ; Zheng LI ; Yin-fa HAN
National Journal of Andrology 2005;11(7):520-525
OBJECTIVETo compare the efficacy and safety of sildenafil plus sertraline with those of sertraline alone in the treatment of premature ejaculation (PE).
METHODSSeventy-two patients with PE but without any obvious organic cause were enrolled in this study. They were randomly divided into Groups A and B of equal number. Group A received 50 mg sertraline daily 4 to 6 hours before planned sexual activity for 12 weeks, and Group B were given 50 mg sertraline daily plus 50 mg sildenafil as needed, 1 hour before planned sexual activity, for 12 weeks. Before and after the treatment, the mean intravaginal ejaculation latency time, the intercourse satisfaction, the mean number of coituses per week and the drug-related side effects were evaluated.
RESULTSThe mean intravaginal ejaculatory latency time was (0.59 +/- 0.12), (3.9 +/- 0.15) minutes (P < 0.001) at baseline and post-treatment in Group A, and (0.56 +/- 0.11), (5.6 +/- 0.12) minutes (P < 0.001) in Group B, improved in both of the 2 groups, but more significantly in Group B (P < 0.05). Before and after the treatment, the mean intercourse satisfaction domain values of the IIEF were (8.9 +/- 1.2), (10.8 +/- 1.1) (P < 0.05) and (8.8 +/- 1.1), (13.8 +/- 1.3) (P < 0.001) in Groups A and B, respectively, significantly greater in Group B than in Group A (P < 0.05) after the treatment; the mean numbers of coituses per week in Groups A and B were (0.9 +/- 0.2), (1.9 +/- 0.3) (P < 0.05) and (1.0 +/- 0.2), (2.7 +/- 0.2) (P <0.001) respectively, significantly larger in Group B (P<0.05) after the treatment. As for the side effects, there was a higher rate of headaches (P < 0.01) and flushing episodes (P < 0.001) in Group B than in Group A.
CONCLUSIONSertraline combined with sildenafil can produce significantly better results than sertraline alone in patients with premature ejaculation. However, the combined treatment is associated with a slight increase in the drug-related side effects.
Adolescent ; Adult ; Drug Therapy, Combination ; Ejaculation ; Genital Diseases, Male ; drug therapy ; Humans ; Male ; Phosphodiesterase Inhibitors ; adverse effects ; therapeutic use ; Piperazines ; administration & dosage ; adverse effects ; Purines ; administration & dosage ; adverse effects ; Sertraline ; administration & dosage ; adverse effects ; Sildenafil Citrate ; Sulfones ; administration & dosage ; adverse effects
6.Optimization of Water Extraction and Alcohol Precipitation Process of Qizhi Yifei Granules by Multi-Index Orthogonal Experimental
Wen-Ping WANG ; Jian NI ; Xin LENG ; Chun-Jing YANG ; Long-Tai YOU ; Yi LIU ; Na SAI ; Xiao LIANG ; Xing-Bin YIN
Chinese Journal of Information on Traditional Chinese Medicine 2018;25(9):71-75
Objective To optimize the extraction and alcohol precipitation process of Qizhi Yifei Granules by multi index orthogonal experiment. Methods With extraction rate of astragaloside in Astragali Radix, quercetin-3-O-β-D-glucose-7-O-β-D-gentian diglucoside in Descurainiae Semen Lepidii Semen and yield rate of dry extract as indexes, the extraction process of Qizhi Yifei Granules was optimized. Taking the retention rate of astragaloside and quercetin-3-O-β-D-glucose-7-O-β-D-gentian diglucoside as indexes, the alcohol precipitation process was optimized. Results The best water extraction process was as follows: adding 10 times amount of water, extracting for 1.5 h, 3 times. The optimum alcohol precipitation process was: concentrated to the relative density of 1.05–1.10 (60 ℃), adding ethanol to 60% and alcohol precipitation. Conclusion The optimized extraction and alcohol precipitation process is stable and feasible, which can provide the basis for the preparation.
7.Discovery of biomarkers related to abnormal lipid metabolism in liver and serum and intervention mechanism of ginsenoside Rb_1 in hyperlipidemia rats based on non-targeted metabolomics.
China Journal of Chinese Materia Medica 2023;48(14):3922-3933
Through the non-targeted metabolomics study of endogenous substances in the liver and serum of hyperlipidemia rats, the biomarkers related to abnormal lipid metabolism in hyperlipidemia rats were found, and the target of ginsenoside Rb_1 in improving hyperlipidemia was explored and its mechanism was elucidated. The content of serum biochemical indexes of rats in each group was detected by the automatic biochemical analyzer. The metabolite profiles of liver tissues and serum of rats were analyzed by HPLC-MS. Principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to compare and analyze the metabolic data in the normal group, the hyperlipidemia group, and the ginsenoside Rb_1 group, and screen potential biomar-kers. The related metabolic pathways were further constructed by KEGG database analysis. The results showed that hyperlipemia induced dyslipidemia in rats, which was alleviated by ginsenoside Rb_1. The non-targeted metabolomics results showed that there were 297 differential metabolites in the liver tissues of hyperlipidemia rats, 294 differential metabolites in the serum samples, and 560 diffe-rential metabolites in the hyperlipidemia rats treated by ginsenoside Rb_1. Perillic acid and N-ornithyl-L-taurine were common metabolites in the liver and serum samples, which could be used as potential biomarkers for ginsenoside Rb_1 in the improvement of hyperlipidemia. As revealed by pathway enrichment in the liver and serum, ginsenoside Rb_1 could participate in the metabolic pathway of choline in both the liver and serum. In addition, ginsenoside Rb_1 also participated in the ABC transporter, alanine, aspartic acid, and glutamate metabolism, protein digestion and absorption, β-alanine metabolism, taurine and hypotaurine metabolism, caffeine metabolism, valine, leucine, and isoleucine biosynthesis, arachidonic acid metabolism, and methionine and cysteine metabolism to improve dyslipidemia in rats.
Rats
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Animals
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Hyperlipidemias/drug therapy*
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Metabolome
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Ginsenosides/metabolism*
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Lipid Metabolism
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Metabolomics/methods*
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Liver/metabolism*
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Biomarkers
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Taurine
8. Effects of overexpression of IncRNA AC079466.1 on apoptosis of NSCLC cells through endoplasmic reticulum stress signaling pathway
Meng-Lin FENG ; Xin-Yi WEI ; Ping WANG ; Leng-Xin DUAN ; Nan-Ya WANG
Chinese Pharmacological Bulletin 2023;39(9):1689-1695
Aim To investigate the expression of IncRNA AC079466. 1 in non-small cell lung cancer (NSCLC) tissues and cells, and the effect of its overexpression on the proliferation, apoptosis, migration and invasion of A549 and H1299 cells. Methods Cancer tissues and corresponding adjacent tissues from 20 NSCLC patients were collected, and the expression of IncRNA AC079466. 1 in tissue and cells was detected by qRT-PCR. AC079466. 1 group was transfected with overexpression plasmid, NC group was transfected with empty plasmid, and no transfection was used in the Blank group. MTT, flow cytometry and Transwell were used to detect the effects of IncRNA AC079466. 1 overexpression on the viability, apoptosis, migration and invasion of A549 and HI299 cells. Western blot was used to detect the effect of overexpression of IncRNA AC079466. 1 on the expression of endoplasmic reticulum stress-related factors GRP78, PERK, eIF2a, ATF4, CHOP, Bax and caspase-3. Results Compared with adjacent tissues, the expression of IncRNA AC079466. 1 in cancer tissues significantly decreased. Compared with HBE cells, the expression of IncRNA AC079466. 1 significantly decreased in A549 and H1299 cells. Compared with the Blank group and NC group, the viability, migration and invasion abilities of A549 and H1299 cells in AC079466. 1 group all markedly decreased, the apoptosis rate apparently increased, and the expressions of endoplasmic reticulum stress-related factors GRP78, p-PERK, eIF2a, ATF4, CHOP, Bax and caspase-3 were significantly up-regulated. Conclusion The overexpression of IncRNA AC079466. 1 significantly inhibits the viability, migration and invasion of A549 and HI299 cells, and promotes cell apoptosis. The mechanism may be related to the promotion of endoplasmic reticulum stress-mediated cell apoptosis.
9.Effects of Smad4 on liver fibrosis and hepatocarcinogenesis in mice treated with CCl4/ethanol.
Xin-bao XU ; Zhen-ping HE ; Xi-sheng LENG ; Zhi-qing LIANG ; Ji-run PENG ; Hong-yi ZHANG ; Hong-yi ZHANG ; Mei XIAO ; Hui ZHANG ; Cheng-li LIU ; Xi-dong ZHANG
Chinese Journal of Hepatology 2010;18(2):119-123
To study the effects of Smad4 on liver fibrosis and hepatocarcinogenesis in mice treated with CCl(4)/ethanol. The wild-type mice (Smad4 +/+) and the Smad4 knockout mice (Smad4 +/-) were injected subcutaneously with carbon tetrachloride(CCl(4))/ethanol twice a week for twenty weeks. The expression of Smad4, TGFbeta1, Smad2, Smad3, Smad6, TIMP1, MMP2 and MMP9 was detected by RT-PCR. In the cirrhotic liver, the expression of Smad4 mRNA was significantly higher than that in the normal liver. Comparing with wild-type mice (Smad4 +/+), the TGFbeta1-Smad4 signaling was markedly attenuated in the Smad4 knockout mice (Smad4 +/-). After induction by CCl(4)/ethanol, the hepatic fibrosis in the Smad4 knockout mice (Smad4 +/-) was obviously alleviated compared with the wild-type mice (Smad4 +/+), and the incidence rate of hepatocarcinogenesis of the former was also lower than that of the latter(32.0% vs 41.9%). These results indicate that knocking out Smad4 can delay the progression of liver fibrosis and liver cancer.
Animals
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Carbon Tetrachloride
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administration & dosage
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Disease Models, Animal
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Ethanol
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administration & dosage
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Female
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Liver Cirrhosis, Experimental
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chemically induced
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metabolism
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pathology
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Liver Neoplasms, Experimental
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chemically induced
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metabolism
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pathology
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Male
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Mice
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Mice, Knockout
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RNA, Messenger
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genetics
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metabolism
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Reverse Transcriptase Polymerase Chain Reaction
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Signal Transduction
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Smad Proteins
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genetics
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metabolism
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Smad4 Protein
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genetics
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metabolism
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Tissue Inhibitor of Metalloproteinase-1
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genetics
;
metabolism
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Transforming Growth Factor beta1
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genetics
;
metabolism
10.Effect of angiotensin converting enzyme inhibitor on the calcium transients and calcium handling proteins in ventricular myocytes from rats with heart failure.
Li-chun WANG ; Hong MA ; Jian-gui HE ; Xin-xue LIAO ; Wen-fang CHEN ; Xiu-yu LENG ; Li MA ; Wei-yi MAI ; Jun TAO ; Wu-tao ZENG ; Jun LIU ; Yu-gang DONG ; An-li TANG ; Chong FENG
Chinese Medical Journal 2005;118(9):731-737
BACKGROUNDChronic heart failure (CHF) is associated with calcium transients and calcium handling proteins. Angiotensin converting enzyme (ACE) inhibitor has been demonstrated to have beneficial effect on CHF. Yet studies addressed to the relationship between ACE inhibitor and calcium transients in CHF are rare. The aim of this study was to investigate the influence of ACE inhibitor (perindopril) on the contractility and calcium transients and calcium handling proteins in ventricular myocytes from rats with experimental heart failure.
METHODSMale Wistar rats were randomized to heart failure group treated with perindopril [CHF-T, 3 mg.kg(-1).d(-1)], heart failure group without treatment (CHF-C) and sham-operated group (PS). Heart failure was induced by abdominal aortic constriction. All groups were further followed up for 12 weeks. Left ventricular myocytes were then isolated. Single cell shortening fraction and [Ca(2+)]i were simultaneously measured by laser scanning confocal microscope under the field stimulation (1.0 Hz). Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot were performed to evaluate the changes of mRNA and protein of Na(+)-Ca(2+) exchanger (NCX1), sarcoplasmic reticulum Ca(2+)-ATPase (SERCA2) and phospholamban (PLB).
RESULTSThe fraction of cell shortening (FS%) and [Ca(2+)]imax (nmol/L) were significantly reduced in group CHF-C compared with group PS (FS%: 7.51 +/- 1.15 vs 13.21 +/- 1.49; [Ca(2+)]i max: 330.85 +/- 50.05 vs 498.16 +/- 14.07; both P < 0.01), and restored at least partially in CHF-T group. In CHF-C group, the left ventricular mRNA of NCX1 and PLB were significantly upregulated in comparing with PS group (RNCX1/beta-Actin: 0.51 +/- 0.12 vs 0.19 +/- 0.06, P < 0.01; RPLB/beta-Actin: 0.26 +/- 0.12 vs 0.20 +/- 0.08, P < 0.05), while SERCA2 mRNA was downregulated (0.48 +/- 0.10 vs 0.80 +/- 0.11, P < 0.01). The mRNA levels of NCX1 and SERCA2 in CHF-T group were between the CHF-C and PS group, and the differences of the latter two groups were significant (all P < 0.05). In CHF-C and CHF-T groups, the protein expression of NCX1 were 1.141 +/- 0.047 and 1.074 +/- 0.081 times of that in PS group respectively (both P < 0.05), and SERCA2 protein levels were 0.803 +/- 0.100 and 0.893 +/- 0.084 times of that in PS group respectively (both P < 0.05). The protein expression of NCX1 and SERCA2 in the CHF-C and CHF-T groups is significantly different (both P < 0.05).
CONCLUSIONACE inhibitor could improve cardiac function of failing heart through directly enhancing the contractility of single cardiomyocyte, and these effects are probably mediated by its roles in preventing the deleterious changes of calcium transients and calcium handling proteins in CHF.
Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Animals ; Calcium ; metabolism ; Calcium-Binding Proteins ; genetics ; Calcium-Transporting ATPases ; genetics ; Heart Failure ; drug therapy ; metabolism ; Heart Ventricles ; drug effects ; Male ; Myocytes, Cardiac ; drug effects ; metabolism ; Perindopril ; pharmacology ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; Sodium-Calcium Exchanger ; genetics