ABSTRACT
The objective of this work was to evaluate the anti-fatigue efficacy of Astragali Radix (AR) from the Shanxi Hengshan area and to reveal possible mechanisms by which it relieves fatigue. Efficacy differences between Guangling (GL) and Hunyuan (HY) AR preparations were compared and evaluated, and an 1H NMR metabolomic technique combined with statistical methods was used to identify the metabolites in different groups of mouse gastrocnemius muscle tissues. The differential metabolites after AR treatments were identified according to VIP and P values and the upstream targets were predicted with the help of Metscape. Cytoscape software was utilized to construct a network map of AR potential anti-fatigue targets. Key differential metabolites were identified based on shared targets and entered into the Metaboanalyst website for pathway enrichment analysis, which led to the preliminary elucidation of the molecular mechanisms. The results showed that intervention with AR can significantly improve the swimming-to-exhaustion time, increase liver glycogen, and reduce urea-nitrogen levels in mice. The difference between GL and HY ARs was relatively small, indicating that the quality of AR produced in the Hengshan area is consistent and stable. The metabolic fingerprints of mouse gastrocnemius muscle tissue extracts were composed of 34 metabolites, and the statistical results showed that 19 differential metabolites were significantly reversed after the Hengshan AR intervention. We found that the anti-fatigue effects of AR in the Shanxi Hengshan area were mainly associated with taurine and hypotaurine metabolism through regulation of GAD1, based on network pharmacological analysis. In conclusion, 1H NMR metabolomic techniques were combined with network pharmacology to compare and evaluate the quality of Hengshan ARs, and further associate the fatigue relieve with the regulation of taurine metabolism. This provides a theoretical basis for the resource utilization of Hengshan ARs and the development of anti-fatigue-related products. The animal experiments in this study followed the regulations of the Animal Ethics Committee of Shanxi University and passed the ethical review of animal experiments (Approval No. SXULL2021028).
ABSTRACT
The pathogenesis of the nephrotic syndrome is complex and the pathological types are diverse, so the minor symptoms in its early phases are difficult to detect. Renal biopsy is the gold indicator for the diagnosis of renal pathology and progression, but poor patient compliance shows, and the optimal treatment time is often delayed. Therefore, the discovery of biomarkers for early diagnosis and disease progression monitoring is of great clinical significance. In this study, doxorubicin-injured podocyte models were used to simulate human kidney disease at different stages of progression. LC-MS-based metabolomic technology combined with statistical methods was used to screen and identify the potential biomarkers associated with early injury or progression of podocytes. The results of cell viability, apoptosis tests and podocyte structural protein analysis showed that the model was successfully constructed, and the degree of podocyte injury was significantly different between the two modeling methods. According to VIP > 1 and P < 0.05 based on the orthogonal partial least squares discriminant analysis (OPLS-DA) model, nine differential metabolites reflecting early podocyte injury and twelve differential metabolites reflecting the injury progression were screened, respectively. ROC analysis was adopted to focus on the potential biomarkers that can reflecting the early podocyte injury including L-tryptophan, guanosine triphosphate (GTP), 5′-thymidylic acid (dTMP) and thymidine, and the biomarkers reflecting the injury progression of podocytes composed of L-phenylalanine, L-tyrosine acid, uridine 5′-diphosphate (UDP) and guanosine 5′-diphosphate (GDP) AUC > 0.85. It indicated that these eight metabolites may have high sensitivity and diagnostic ability. This study provides a reference for the research on biomarkers of progressive diseases.
ABSTRACT
The quality of Astragali Radix (AR) was closely related to the growth period. However, the current commodity grades of AR were only divided by diameter but not directly related to the growth period, which leads to the contradiction between the grade standard and the quality evaluation index. Therefore, solving this problem will be the key for the quality evaluation of AR. The present study established a potential quality evaluation approach for the absolute growth years' wild Astragali Radix (WAR) and transplanted Astragali Radix (TAR) based on the chemical components and anti-heart failure efficacy through adopting a bare-handed sections approach to rapidly identify the growth years of WAR. In this study, the absolute growth years of WAR were obtained by identifying the growth rings of 1-6 growth years root through the methods. The contents of flavonoids and saponins in 2-6 growth years' WAR were determined by HPLC-UV-ELSD. The contents of 12 chemical components and the anti-fatigue failure effects of WAR (4-year-old) and TAR were compared on rat models of heart failure induced by doxorubicin. Meanwhile, NMR-based untargeted metabolomics studies were performed to investigate the regulative effects of WAR and TAR. The result shows that the numbers of growth rings were consistent with the actual growth periods of AR. The HPLC-UV-ELSD determination indicated that the content of total flavonoids in WAR was significantly higher than that in TAR. Pharmacodynamics analysis revealed that the effects of WAR on cardiac function parameters (EF, FS and LVIDs), contents of serum CK and BNP were superior to those of TAR. 13 metabolites of heart were identified that had a higher rate of change in WAR group than TAR. Overall, a rapid identification method for the growth years of WAR was established, and the fact that WAR were significantly better than TAR in the heart failure rats was first proved in the paper. This study provided a scientific basis for establishing a novel commodity specification and grade of AR for clinical rational drug use.
ABSTRACT
The effective material basis of traditional Chinese medicine(TCM) is an indispensable part of studies on TCM, and each technology has its advantages and disadvantages. The target constituent knock-out/knock-in technology has attracted much attention since it was proposed because of its unique advantages of regarding the extract of the formula as a whole, which can better reflect the characteristics of multi-component and multi-target integration and regulation of TCM. This method investigated the contribution of target constituent to the overall efficacy of a TCM by analyzing the changes in efficacy of the remaining formula before and after knock-out/knock-in of the target constitution. The application of this model not only facilitates studies of the effective constituents of TCM, but also help to develop the quality control standard of TCM. However, the application of this model is restricted due to the limitation of target constituent separation technology. By reviewing the literatures in recent years, this study summarized the research process and application of this method for a reference.
Subject(s)
Animals , Drugs, Chinese Herbal/pharmacology , Gene Knock-In Techniques , Gene Knockout Techniques , Medicine, Chinese Traditional , Quality ControlABSTRACT
Ziziphi Spinosae Semen (ZSS) has been used for treatment of insomnia in China for centuries. To reveal the influence of insomnia on the levels of the neurotransmitters including serotonin (5-HT), glutamic acid (Glu), γ-aminobutyric acid (GABA), noradrenaline (NE) and dopamine (DA), and to study the role of ZSS aqueous extract in the treatment of insomnia, an UPLC-ESI- MS/MS method was developed and validated for simultaneous determination of five neurotransmitters in the rat brain. The brain samples were pretreated by one-step direct protein precipitation with acetonitrile. The analytes were detected in positive mode with multiple reaction monitoring (MRM) and the procedure was completed in less than 10 min. The method showed a good linearity (R > 0.9967) with the other validation parameters were within acceptance range. The results indicated that the concentration of 5-HT, GABA and DA is significantly lower (P < 0.01) in para-chlorophenylalanine (PCPA)-induced insomnia rat model group, while Glu and NE significantly higher than those in control group (P < 0.01). Treatment with ZSS aqueous extract (4 or 8 g·kg·d for seven days) could ameliorate the symptoms of insomnia by significantly changing the levels of the neurotransmitter parameters mentioned above. The data obtained in this study demonstrate that ZSS aqueous extract could ameliorate the symptoms of insomnia by modulating the levels of monoamines and amino acid neurotransmitters in the brain.
ABSTRACT
Objective: To explore the receiuer operating characteristic(ROC) curve mathematical statistics method to establish the content limit of isoflavones glucoside contents in Astragali Radix,in order to distinguish the difference in isoflavones glucoside content between Hengshan wild-simulated Astragali Radix in Shanxi and cultivated Astragali Radix in Gansu. Method: A total of 225 samples of 45 batches of wild-simulated Astragali Radix in Hengshan and 210 samples of 42 batches of cultivated Astragali Radix in Gansu were selected as research objects,and isoflavones glucoside contents of these samples were determined by using the method in the edition 2015 Chinese Pharmacopoeia. Data was statistically analyzed, and the ROC curve was used to determine the critical value of wild-simulated Astragali Radix in Hengshan and transplanted Astragali Radix in Gansu. Result: The average contents of isoflavones glucoside in imitative wild Astragali Radix in Henshan and cultivated Astragali Radix in Gansu were 0.107%and 0.039%respectively,with statistically significant differences(PConclusion: ROC curve can be used to distinguish the critical value of the wild-simulated Astragali Radix in Hengshan and cultivated Astragali Radix in Gansu. The results reflected the regularity of "high quality and good price" in the market. The research provided a theoretical basis for the establishment of standard of high-quality Chinese herbal medicines.
ABSTRACT
A rapid and accurate method for determination of astragaloside Ⅳ was established,which was further applied to determine the contents of astragaloside Ⅳ in 87 batches of different origin and different grade of Astragali Radix. The ROC curve was used to analyze the contents of astragaloside Ⅳ in different origin. Simultaneous contents of astragaloside Ⅳ in different grade were compared with chemometrics. HPLC-ELSD method was used to determine the contents of astragaloside Ⅳ. A Vensil MP C18 column( 4. 6 mm×250 mm,5 μm) was used with acetonitrile-water( 32 ∶68) as the mobile phase at a flow rateof 1 m L·min-1. The column temperature was 25 ℃ with ELSD parameters as follows: gas flow rate was 2. 5 L·min-1,the drift tube heating temperature was set to 105 ℃,and the gain value was 4. 0. The optimized method avoided the problem that the consumable quality unstable and the recovery rate was not high. The contents determined by the optimized method were higher than the pharmacopoeia method,with less time and high recovery rate. The ROC curve analysis showed that there was no significant difference of contents of astragaloside Ⅳ between the top grade of Shanxi wild-simulated Astragali Radix top and the first grade of Gansu cultivated Astragali Radix. The contents of astragaloside Ⅳ in the second,third and fourth grade of Shanxi wild-simulated Astragali Radix was significantly higher than those of produced from Gansu.There was a significant negative correlation between the contents of astragaloside Ⅳ and grade in Shanxi Astragali Radix. While there was no correlation for Gansu Astragali Radix. This study provided the basis for the quality grade standard of Astragali Radix.
Subject(s)
Astragalus Plant , Chemistry , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Saponins , TriterpenesABSTRACT
We studied the effect of aqueous extract from Huang qi on gene expression profile of doxorubicin induced nephropathy in rats, and explored the molecular mechanism of the intervention. The gene expression profiles of control group, model group and aqueous extract from Huang qi group were detected by using transcriptome sequencing technique. The differentially expressed genes (DEGs) were screened by STEM trend analysis software. GO function enrichment and KEGG pathway analysis were performed for DEGs, and the gene expression level was verified by real-time fluorescence quantitative PCR (RT-qPCR). The results showed that, compared with the control group, 432 DEGs were obtained in doxorubicin nephropathy model group; compared with the model group, 811 DEGs were obtained due to aqueous extract of Huang qi. The results of GO function enrichment and KEGG enrichment analysis indicated that PI3K-AKT pathway (Col6a6, Nr4a1, Sgk1, Gng7) and lipid metabolism-related genes (Cpt1b, Pcsk9, Abca1, Ascm5) were the key pathways and genes in the treatment of doxorubicin induced nephropathy by aqueous extract from Huang qi, which played a protective role in kidney. In conclusion, the molecular mechanism of aqueous extract from Huang qi in protection against doxorubicin induced nephropathy rats is closely related to apoptosis-related genes and lipid metabolism-related genes, suggesting for the need of follow-up study for key gene validation and mechanism of action of aqueous extract from Huang qi for prevention of doxorubicin induced nephropathy.
ABSTRACT
Objective To determine the optimal model for the evaluation of pharmacodynamics of Fangji Huangqi Decoction. Methods By changing the doses and times of doxorubicin and the intervention of Fangji Huangqi Decoction and a series of test drugs, the effects of different doses and times of doxorubicin on the model were explored, and body weight, organism index, urine protein quantitation, renal histopathology results, and serum biochemistry can be regarded as evaluation indexes. Results For M1 (5 + 2) mg/kg model, the mortality rate of rats was higher. For M2 (6 mg/kg) model, the mortality rate was up to 20%, and the indicators reflecting renal function did not callback, but instead it showed much higher levels than the model group after receiving the test drug. For M3 (4 + 2) mg/kg model, the callback of part of the indexes, such as lung and kidney organ index, serum total triglyceride, and total protein levels, occurred after receiving the test drug, but the curative effect is not ideal. For M4 or M4’ (4 + 1) mg/kg model, the quantitative level of urinary protein in model rats was significantly different from that in the control at the 14th d, and the sustained and stable growth trend was observed; Moreover, the indicators mentioned above have different degrees of callback with high-, middle-, and low dose of Fangji Huangqi Decoction. Conclusion The final model weight of the rats was determined to be about 300 g, and the cumulative dose of doxorubicin was 5 mg/kg tail intravenous injection of doxorubicin was adopted as follows: on the 1st d of the experiment, 4 mg/kg doxorubicin was injected, and followed another 1 mg/kg at intervals of 1 week, which was the best way to evaluate the pharmacological effect of the related drugs derived from the Fangji Huangqi Decoction and the middle dose was the best.
ABSTRACT
This study was designed to explore the intervention of muscle fatigue in rats with Astragali Radix using 1H NMR metabolomics methods. The fatigue model was induced in rats by forced swimming plus food restriction, and the effects of Astragali Radix (3, 6 and 12 g·kg-1) were investigated using the exhaustive time of rat swimming. After 3 weeks, the gastrocnemius was collected for 1H NMR detection, and the anti-fatigue effects of Astragali Radix were explored using multivariate statistical analysis. Astragali Radix significantly improved the exhaustive swimming time of rats. Compared with control group, the levels of isoleucine, leucine, creatine, phosphatidylcholine, trimethylamine oxide, taurine, guanidinoacetate, AMP, inosine, histidine, hypoxanthine, anserine in rat gastrocnemius of model group were increased. While the levels of lactate, acetone, choline, glycerophosphocholine, glycine were decreased. These 6, 11, 5 potential biomarkers could be reversely regulated by treatment with Astragali Radix (high dose, middle dose, low dose), respectively. Metabolomics analysis revealed that Astragali Radix has a certain anti-fatigue effects and the mechanism may be related to regulation of amino metabolism.
ABSTRACT
This study was designed to explore the mechanism of total flavonoids of Astragali Radix (TFA) in treating nephrotic syndrome through establishing the active components-targets network and protein-protein interaction (PPI) networks and analyzing the functions and pathways involved in the targets. The main active ingredients of TFA were obtained by 1H NMR and LC-MS, TCMSP and TCMID database. PharmMapper, SEA, SIB, HOME-NCBI-GENE, GeneCards and OMIM were used to predict and screen the active components of TFA. The Cytoscape software was used to construct the active components-targets network and protein-protein interactions network. The relation between the main active ingredients and targets were validated by Systems Dock Web Site. The GO and KEGG pathways involved in the targets were analyzed by ClueGO software. The target organ distribution was assigned by the BioGPS database. The results showed that 29 active components and 50 targets of TFA were screened and predicted. The network results showed that the TFA were mainly involved in biological processes such as inflammatory reaction process, oxidative stress process,apoptosis and autophagy, and played a role in the regulation of AGE-RAGE, PI3K/Akt, VEGF, IL-17 and MAPK signaling pathways to treat the nephrotic syndrome. This study reflects the characteristics of multi-components, multi-targets and multi-pathways of TFA, which provides new ideas and clues for further research on the mechanism of anti-nephrotic syndrome effects of TFA.
ABSTRACT
Astragali Radix (AR) is one of the most popular herbal medicines in traditional Chinese medicine (TCM). Wild AR is believed to be of high quality, and substitution with cultivated AR is frequently encountered in the market. In the present study, two types of ARs (wild and cultivated) from Astragalus membranaceus (Fisch.) Bge. and A. membranaceus var. mongholicus (Bge.) Hsiao, growing in different regions of China, were analyzed by NMR profiling coupled with multivariate analysis. Results showed that both could be differentiated successfully and cultivation patterns or growing years might have greater impact on the metabolite compositions than the variety; the metabolites responsible for the separation were identified. In addition, three extraction methods were compared and the method (M1) was used for further analysis. In M1, the extraction solvent composed of water, methanol, and chloroform in the ratio of 1 : 1 : 2 was used to obtain the aqueous methanol (upper layer) and chloroform (lower layer) fractions, respectively, showing the best separation. The differential metabolites among different methods were also revealed. Moreover, the sucrose/glucose ratio could be used as a simple index to differentiate wild and cultivated AR. Meanwhile, the changes of correlation pattern among the differential metabolites of the two varieties were found. The work demonstrated that NMR-based non-targeted profiling approach, combined with multivariate statistical analysis, can be used as a powerful tool for differentiating AR of different cultivation types or growing years.
Subject(s)
Astragalus propinquus , Chemistry , Classification , Metabolism , China , Magnetic Resonance Spectroscopy , Methods , Metabolomics , Plant Extracts , Chemistry , Metabolism , Plant Roots , Chemistry , Classification , Plants, Medicinal , Chemistry , MetabolismABSTRACT
To investigate the difference of Gegen Qinlian Decoction (GQD) piece and boiled powder in the treatment of type 2 diabetes (T2DM), the characteristic of overall metabolite profile was examined in the serum of T2DM rats with 1H NMR-based metabolomics combined with the multivariate statistical analysis. A rat model of T2DM was established by feeding of high glucose and high fat diet followed by a streptozotocin (STZ) treatment. The general condition, body weight and fasting blood glucose (FBG) of rats were monitored. GQD piece and boiled powder exhibited activities in the improvement of these parameters. The results of the principal component analysis showed that there was a significant difference in the metabolic profile of the normal control group, the model group, the positive group, the herbal decoction group and the boiled powder group. Totally 15 potential biomarkers were identified by OPLS-DA binding univariate analysis. Compared with normal control group, the serum samples of T2DM showed a higher level of 3-HB, TMAO, glycine, β-glucose and α-glucose accompanied by lower level of lactate, VLDL, acetate, glutamate, methionine, glutamine, pyruvate, creatine, choline and glycerol. The above results also demonstrated that both piece and boiled powder of GQD could restore 14 of these markers. These results suggested that the disrupted metabolic pathways including energy metabolism, lipid metabolism and amino acid metabolism, were restored by GQD piece and boiled powder. The two formula did not show a significant difference. The results of this study provide experimental data and theoretical basis for the equal activities of GQD piece and boiled powder in clinical application.
ABSTRACT
Shanxi, a traditional production area to produce genuine Astragali Radix of high quality, has experienced major changes in the pattern of resources. This area once accounted for half of Astragali Radix industry, but now only serves as the largest supply area of traditional wild Astragali Radix. Furthermore, the strategic position of Shanxi Astragali Radix industry will become more prominent and more important to economic and social development in face of the diversity of market demands, especially for the strong demands of high-end Astragali Radix. In addition, Astragalus industry involves the simultaneous development of the first, second and tertiary industries in many areas, and it is typical and representative in the traditional Chinese medicine industry development. However, the application and industrial development of Shanxi Astragali Radix have been restricted due to the problems such as blind promotion of transplanting cultivation technology, and lack of science and technology including efficacy investigation, safety evaluation, standardization and controllability studies. Therefore, we would analyze the production history, resource structure, the current situation and progress of industry development, scientific research foundation and existing problem in this paper, and put forward countermeasures for development and technical innovation in order to make Astragali Radix industry bigger and stronger through innovation-driven and make benefits for demos. This thought provides a reference for the exploratory development of other large varieties of Chinese medicinal materials.
ABSTRACT
The study established the 1H NMR-based fingerprinting and analyzed 8 batches of Huangqi injection solution. 1H NMR-based fingerprinting of both primary and secondary metabolites of Huangqi injection were established, and the 1H-1H chemical shift correlation spectroscopy (COSY) and 1H-13C chemical shift correlation spectroscopy (HSQC) were used to identify the chemical components in Huangqi injection solution. Coupled with similarity analysis and relative content determination, 8 batches of Huangqi injection solution were analyzed. Twenty-five metabolites (both primary and secondary) were identified, and the significant differences were found in the chemical composition among these Huangqi samples. The content and content variation of the primary metabolites were much higher than those of the secondary metabolites, which was the major cause of the uniformity of the Huangqi injections. The results on the quality variations of Huangqi injections in this study will serve as a basis for improving the quality control.
ABSTRACT
Growth year is one of the important factors for the quality of Polygala tenufolia. In this study, primary metabolites and secondary metabolites were compared in 1, 2 and 3 years old P. tenufolia cultivated in Shaanxi Heyang. The samples were subjected to ultra-high performance liquid chromatography (UPLC)-quadrupole time-of-flight mass spectrometry (Q-TOF MS) and nuclear magnetic resonance (NMR) analysis, and the obtained data were analyzed using principal component analysis (PCA) and other statistical analysis methods. In addition, content and correlation of different metabolites were also calculated. The results showed no significance between main component contents in 2 year-old and 3 year-old P. Tenufolia, but 1 year-old was statistically different. The contents of primary metabolites, such as fructose, sucrose, and choline increased as time goes on, while glycine and raffinose decreased. The contents of secondary metabolites, such as onjisaponin Fg, polygalasaponin XXVIII, polygalasaponin XXXII increased, while polygalaxanthone III and parts of oligosaccharide multi-ester including tenuifoliose A, tenuifoliose C, tenuifoliose C2 and tenuifoliose H decreased with the extension of the growth years. Growth years has important impact on the quality of P. tenuifolia and the existing growing years of commodity P. tenuifolia have its scientific evidence. This study supplied a new method for the quality evaluation of Chinese medicinal materials.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Magnetic Resonance Spectroscopy , Mass Spectrometry , Metabolomics , Oligosaccharides , Plants, Medicinal , Chemistry , Polygala , Chemistry , Quality ControlABSTRACT
<p><b>OBJECTIVE</b>To explore the role of the phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt) pathway in silica-induced α-SMA (α smooth muscle actin) expression in HEB (human bronchial epithelial) cell.</p><p><b>METHODS</b>The cultured HBE cells were divided into 5 groups: control, silica, PI3K inhibitor (Ly294002), both PI3K inhibitor (Ly294002) and silica at the same time and the inhibitor 24 h ahead of silica. The final concentrations of PI3K inhibitor and silica were 10 µmol/L and 100 µg/ml, respectively. Western blots were used to detect protein expressions of Akt, p-Akt, TGF-β and α-SMA. The location and expression of α-SMA were measured by immunofluorescence assay.</p><p><b>RESULTS</b>HBE cell line exposed to silica can induce Akt phosphorylation, in which expressions of p-Akt were up regulated 1 times at 48 and the highest at 72 h. The expressions of TGFβ increased remarkably at 12 h and the peak at 48 h after silica exposure, while the expressions of α-SMA increased at 24 h and the highest at 72 h. However, the PI3K inhibitor (Ly294002) significantly down regulated α-SMA expression. When the cell line exposed to the PI3K inhibitor ahead of silica 24 h, the expressions of p-Akt and α-SMA were more remarkably down regulated which were decreased 1.5 times and 7.6 times respectively compare to silica exposure group. But no significant changes were found for TGFβ expressions. The immunofluorescence assay showed that silica can induce α-SMA expression, which located in cytoplasma, and PI3K inhibitor can decrease the expression.</p><p><b>CONCLUSIONS</b>Silica induced α-SMA expression in HBE cell line is by targeting the PI3K/Akt pathway and PI3K inhibitor can repress α-SMA expression.</p>
Subject(s)
Humans , Actins , Metabolism , Cell Line , Chromones , Pharmacology , Epithelial Cells , Cell Biology , Metabolism , Morpholines , Pharmacology , Phosphatidylinositol 3-Kinases , Metabolism , Phosphorylation , Proto-Oncogene Proteins c-akt , Metabolism , Signal Transduction , Silicon Dioxide , Toxicity , Transforming Growth Factor beta , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the susceptibility to carbon tetrachloride and benzene in offspring of expanded simple tandem repeats (ESTR) mutation mice exposed to formaldehyde (FA).</p><p><b>METHODS</b>F5 and F10 offspring (200 mg/m3 x 2 hours) served as H group and ICR mice were used as control group (group C). The F5 and F10 offspring were exposed to 10 ml/kg carbon tetrachloride at the doses of 0.05%, 0.50% or 5.00% for 24 hours, respectively or 500 or 1000 mg/kg benzene for 24 hours, respectively by intraperitoneal injection. Serum alanine transaminase (ALT), aspartate transaminase (AST) and the hepatic superoxide dismutase (SOD) or malondialdehyde (MDA) were detected; also the hepatic pathological changes were observed under light microscope; the micronucleus in sternum bone marrow cells as the biomarker of benzene blood toxicity were measured.</p><p><b>RESULTS</b>ALT and AST activities in group C of F5 mice exposed to 0.50% and 5.00% CCl4, ALT in groups C and H of F10 mice exposed to 0.05%, 0.50%, 5.00% CCl4, AST in groups C and H of F10 mice exposed to 0.50% and 5.00% CCl4 were significantly higher than those in controls, respectively (P<0.05); as compared to the control, hepatic SOD activities in group C of F5 and F10 mice exposed to 0.50% and 5.00% CCl4, in group H of F5 mice exposed to 0.50% and 5.00% CCl4 and F10 mice exposed to 5.00% CCl4 were significantly reduced, respectively (P<0.05); however, MDA contents in group C of F10 mice exposed to 0.50% and 5.00% CCl4, in group H of F5 mice exposed to 0.05% and 0.50%, 5.00% CCl4 and F10 mice exposed to 0.50% and 5.00% CCl4 were significantly increased than those in control group, respectively (P<0.05). The susceptibility to CCl4 in ESTR mutation F5 mice exposed to FA was significantly higher than that in control F5 mice, but the susceptibility to CCl4 in ESTR mutation F10 mice exposed to FA was significantly lower than that in control F10 mice. The histopathological examination showed that the injury of hepatocytes in C and H groups significantly increased CCl4 doses, and the injury of hepatocytes in H group was higher than that in C group. The micronuclear rates in C and H group mice exposed to benzene(500 mg/kg C group, F5 and F10 mice; 1000 mg/kg C group, F5 and F10 mice; 500 mg/kg H group, F5 and F10 mice; 1000 mg/kg C group, F5 and F10 mice) were 5.88 per thousand +/- 4.55 per thousand, 8.25 per thousand +/- 2.06 per thousand, 7.50 per thousand +/- 6.99 per thousand, 10.67 per thousand +/- 1.16 per thousand, 7.88 per thousand +/- 3.09 per thousand, 9.20 per thousand +/- 1.30 per thousand, 9.63 per thousand +/- 4.34 per thousand and 13.33 per thousand +/- 2.08 per thousand, respectively, which were significantly higher than those (1.13 per thousand +/- 0.35 per thousand, 1.20 per thousand +/- 0.82 per thousand, 1.25 per thousand +/- 0.46 per thousand, 1.33 per thousand +/- 1.03 per thousand) in the solvent control group (P<0.05 or P<0.01).</p><p><b>CONCLUSION</b>FA could result in the change of susceptibility to CCl4 and benzene in offspring of ESTR mutation mice. ESTR mutation may be a biomarker of the susceptibility to chemicals, but the molecular mechanisms should be investigated in the future.</p>
Subject(s)
Animals , Female , Male , Mice , Alanine Transaminase , Metabolism , Aspartate Aminotransferases , Metabolism , Benzene , Toxicity , Carbon Tetrachloride , Toxicity , Chemical and Drug Induced Liver Injury , Environmental Exposure , Formaldehyde , Toxicity , Genetic Predisposition to Disease , Liver , Pathology , Malondialdehyde , Metabolism , Mice, Inbred ICR , Mutation , Superoxide Dismutase , Metabolism , Tandem Repeat Sequences , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To observe the effects of Bushen Zhuyun Recipe (BZR) on protein expressions of estrogen receptor (ER), progesterone receptor (PR) and integrin alpha5 and beta3 in endometrium of rats at the implantation stage, for exploring the possible mechanism of the recipe in treating luteal phase defect (LPD) infertility.</p><p><b>METHODS</b>Female SD rats were randomly divided into 6 groups, the blank group, the model group, the WM group treated by Western medicine, and the three BZR groups treated by low-, middle- and high-dose BZR respectively. Rats were made to pregnancy and sacrificed at the implantation stage, their middle segment of uterus, about 1 cm in length was gotten for detecting the protein expressions by Western blot. Results The protein expressions of endometrial ER and PR were significantly higher, while those of integrin alpha5 and beta3 were significantly lower than those of the control group (P < 0.05). The protein expressions of endometrial ER and PR were significantly lower, but those of integrin alpha5 and integrin beta3 were higher in rats treated by middle- and high- dose BZR than those in model rats (P < 0.05).</p><p><b>CONCLUSION</b>BZR can raise the receptivity of rats' endometrium through down-regulating the expressions of ER, PR and increasing the protein expression of integrin alpha5 and beta3 in endometrium and thus to enhance the pregnant rate.</p>
Subject(s)
Animals , Female , Pregnancy , Rats , Drugs, Chinese Herbal , Pharmacology , Embryo Implantation , Physiology , Endometrium , Metabolism , Integrin alphaV , Metabolism , Integrin beta3 , Metabolism , Rats, Sprague-Dawley , Receptors, Estrogen , Metabolism , Receptors, Progesterone , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To construct antisense c-Jun N-terminal kinase 1 (JNK1) eukaryotic fluorescent expressing vector and JNK1-/- human embryo lung fibroblasts cell line.</p><p><b>METHODS</b>Trizol reagent was used to extract total RNA in HELF. The proper primers of JNK1 were chosen and synthesized. RT-PCR and gene recombinant techniques were used to construct the fragment of JNK1. After purification, the PCR products were cut, and JNK1 were inserted reversely into eukaryotic fluorescent expressing vector pEGFP-C1. Enzyme-cutting and DNA auto-sequencing were used to prove the successful construction of JNK1 eukaryotic expressing vector. Then plasmids were extracted and transfected into HELF cells and screen by G418 24 h later. Monoclone was chosen and cultured. Fluorescent imaging and Western blot were used to identify the JNK1-/- HELF cell line.</p><p><b>RESULTS</b>Sequence analysis of pEGFP-C1-as JNK1 plasmids was same as expected. The expression level of JNK1 was inhibited markedly.</p><p><b>CONCLUSION</b>Construction of antisense JNK1 eukaryotic fluorescent expressing vectors and JNK1-/- HELF cell line is successful.</p>