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1.
J Asian Nat Prod Res ; : 1-13, 2024 Jul 03.
Article de Anglais | MEDLINE | ID: mdl-38958633

RÉSUMÉ

Sesquilignans PD is a natural phenylpropanoid compound that was isolated from Zanthoxylum nitidum var. tomentosum. In this study, we assessed the antitumor effect of PD on SK-Hep-1 and HepG2 cells and the underlying molecular mechanisms. The results revealed that PD markedly inhibited the proliferation and migration of both liver cancer cells. Moreover, PD induced apoptosis, autophagy, and reactive oxygen species (ROS) production in liver cancer cells. Notably, PD increased the protein levels of p-p38 MAPK and p-ERK1/2 in liver cancer cells. This is the first report on the anticancer effect of PD, which is mediated via increased ROS production and MAPK signaling activation.

2.
Fitoterapia ; 173: 105791, 2024 Mar.
Article de Anglais | MEDLINE | ID: mdl-38159614

RÉSUMÉ

Eight undescribed alkaloids named corydalisine D-K (1-7), including one isoquinoline benzopyranone alkaloid (1), one benzocyclopentanone alkaloid (2), four benzofuranone alkaloids (3, 4, and 5a/5b) and two protoberberine alkaloids (6 and 7), along with fourteen known ones, were isolated from the Corydalis saxicola. Their structures, including absolute configurations, were unambiguously identified using spectroscopic techniques, single-crystal X-ray diffraction and electron circular dichroism calculation. Compounds 2, 14 and 21 exhibit antiproliferative activity against five cancer cell lines. The aporphine alkaloid demethylsonodione (compound 14), which exhibited the best activity (IC50 = 3.68 ± 0.25 µM), was subjected to further investigation to determine its mechanism of action against the T24 cell line. The molecular mechanism was related to the arrest of cell cycle S-phase, inhibition of CDK2 expression, accumulation of reactive oxygen species (ROS), induction of cell apoptosis, inhibition of cell migration, and activation of p38 MAPK signaling pathway. The results indicated that 14 could be used as a potential candidate agent for further development of anti-bladder transitional cell carcinoma.


Sujet(s)
Alcaloïdes , Antinéoplasiques , Corydalis , Tumeurs , Corydalis/composition chimique , Structure moléculaire , Alcaloïdes/pharmacologie , Alcaloïdes/composition chimique , Extraits de plantes/composition chimique , Antinéoplasiques/pharmacologie , Dichroïsme circulaire
3.
Bioorg Med Chem ; 80: 117176, 2023 02 15.
Article de Anglais | MEDLINE | ID: mdl-36709571

RÉSUMÉ

A series of 2-oximino-2-indolylacetamide derivatives were designed, synthesized and evaluated for their antitumour effects. Among them, 4d exhibited the most potent antiproliferative effect in vitro on the tested human cancer cells. Additionally, 4d significantly induced cell apoptosis, caused mitochondrial dysfunction, promoted Bax, cleaved-PARP and p53 expression and inhibited Bcl-2 expression in 5-8F cells. Moreover, 4d remarkably promoted autophagosome formation, leading to cell apoptosis. Further investigation indicated that 4d could trigger cell death through cell ferroptosis, including increased ROS generation and lipid peroxidation and decreased glutathione peroxidase 4 (GPx4) expression and glutathione (GSH) levels. More importantly, 4d induced 5-8F cell death by activating ROS/MAPK and inhibiting the AKT/mTOR and STAT3 signalling pathways. Interestingly, 4d significantly suppressed tumour growth in a 5-8F cell xenograft model without obvious toxicity to mice. Overall, these results demonstrate that 4d may be a potential compound for cancer therapy.


Sujet(s)
Antinéoplasiques , Ferroptose , Humains , Animaux , Souris , Espèces réactives de l'oxygène/métabolisme , Apoptose , Antinéoplasiques/pharmacologie , Glutathion/métabolisme , Autophagie
4.
Phytochemistry ; 205: 113476, 2023 Jan.
Article de Anglais | MEDLINE | ID: mdl-36265658

RÉSUMÉ

Eleven previously undescribed alkaloids, including three pairs of enantiomers nitidumalkaloids A-C, a pair of scalemic mixtures nitidumalkaloid D and three optically pure or achiral alkaloids, nitidumalkaloids E-G, along with 20 known alkaloids, were isolated from an ethanolic extract of the whole Zanthoxylum nitidum (Roxb.) DC plant. The chemical structures of the alkaloids were elucidated using a combination of comprehensive nuclear magnetic resonance (NMR) and high-resolution electro-spray ionization mass spectrometry (HR-ESI-MS) analyses. The configuration of the stereogenic centers of all undescribed compounds was precisely established based on single-crystal X-ray diffraction and electronic circular dichroism (ECD) calculations. Racemic mixtures of nitidumalkaloids A-D were purified, and their enantiomers were analyzed via chiral-phase high-performance liquid chromatography with electrochemical detection measurements (HPLC-ECD). Twelve compounds exhibited significant antiproliferative activities against a panel of cancer cell lines. Further studies were designed to investigate the underlying molecular mechanism of (1'S, 6R)-nitidumalkaloid B, which was the most active antiproliferative agent against human cancer A549 cells. G2/M cell cycle arrest, induction of apoptosis, and suppression of the Wnt/ß-catenin signaling pathway were in part associated with the antiproliferative activity of (1'S, 6R)-nitidumalkaloid B. Moreover, (1'S, 6R)-nitidumalkaloid B inhibited cell migration by downregulating the epithelial-mesenchymal transition process in A549 cells. These data suggest that the antiproliferation activity of (1'S, 6R)-nitidumalkaloid B was correlated with the stereoselectivity of the stereoisomers, and (1'S, 6R)-nitidumalkaloid B was prioritized as a potential leading compound for the management of aggressive human non-small-cell lung cancer (NSCLC) from natural products.


Sujet(s)
Carcinome pulmonaire non à petites cellules , Tumeurs du poumon , Zanthoxylum , Humains , Isoquinoléines , Lignée cellulaire
5.
Fitoterapia ; 164: 105381, 2023 Jan.
Article de Anglais | MEDLINE | ID: mdl-36496049

RÉSUMÉ

Five pairs of new racemic alkamides (1a/1b and 4a/4b-7a/7b) and two new achiral derivatives (2-3), as well as five known ones (8-12), were purified from the 95% EtOH extract of Zanthoxylum nitidum. Their structures were elucidated based on spectroscopic analyses (NMR and HR-ESI-MS), electronic circular dichroism (ECD) and NMR calculations. The enantiomeric separation was successfully achieved by chiral-phase HPLC-ECD measurements. Among all the isolates, compounds 2, 3, and 10 showed inhibitory effects against five human cancer cell lines, with IC50 values in range of 18.51-48.03 µM.


Sujet(s)
Zanthoxylum , Humains , Structure moléculaire , Zanthoxylum/composition chimique , Spectroscopie par résonance magnétique , Dichroïsme circulaire
6.
Fitoterapia ; 162: 105289, 2022 Oct.
Article de Anglais | MEDLINE | ID: mdl-36058472

RÉSUMÉ

The chemical investigation on Corydalis balansae resulted in the isolation of three previous undescribed compounds (1, 10, and 11) and 17 known compounds. Compound 1 and 2 were obtained as two lignanamide dimers, and compound 11 had a spiro [benzofuranone-benzazepine] skeleton, which was found in Corydalis for the first time. The structures of new compound were determined by the detailed analysis of 1D/2D NMR, UV, and IR data. Absolute configurations of compounds 10 and 11 were defined by their crystal X-ray diffraction data and calculations of electronic circular dichroism (ECD). The CCK-8 method was used to assay the inhibition effect of all the compounds on the growth of Hela, MGC-803, A549, and HepG2 cancer cells. Compound 2, 13, and 14 showed moderate inhibitory activity against the tested cell lines. Compound 2 exhibited potential antitumor activity against MGC-803 cells with an IC50 value of 20.8 µM, while the positive control etoposide was 17.3 µM. Furthermore, results from the cellular-mechanism investigation indicated that compound 2 could induce S-phase cell-cycle arrest and MGC-803 cells apoptosis, which was triggered by the up-regulation of PARP1, caspase-3 and -9, Bax, and down-regulation of Bcl-2. The 2-induced strong apoptosis indicated that compound 2 had good potential as an antitumor lead compound.


Sujet(s)
Alcaloïdes , Corydalis , Alcaloïdes/composition chimique , Alcaloïdes/pharmacologie , Benzazépines , Caspase-3 , Corydalis/composition chimique , Étoposide , Structure moléculaire , Protéine Bax
7.
Chem Biodivers ; 19(7): e202200449, 2022 Jul.
Article de Anglais | MEDLINE | ID: mdl-35680557

RÉSUMÉ

Zanthoxylum nitidum (Roxb.) DC., is one of Guangxi's characteristic national medicines, and is the classic Laoban medicine of Yao people "Ru Shan Hu" and Zhuang medicine "Liang Bei Zhen". It has been used as an anti-inflammatory, analgesic and haemostatic medicine for thousands of years. In this study, four new sesquiterpenoids (1-4), along with six previously described coumarins (5-10), were isolated from 95 % EtOH extract of Zanthoxylum nitidum. Comprehensive spectroscopic analyses (NMR and HR-ESI-MS) were used to elucidate the structures of these isolates. The absolute configurations of nitidumine A-D (1-4) were established by electronic circular dichroism (ECD). Their cytotoxicity of all the isolates against five cancer cell lines (T24, HeLa, MGC-803, A549, and HepG2) was evaluated by MTT experiment and found not to be cytotoxicity.


Sujet(s)
Médicaments issus de plantes chinoises , Sesquiterpènes , Zanthoxylum , Chine , Coumarines/pharmacologie , Médicaments issus de plantes chinoises/composition chimique , Humains , Structure moléculaire , Sesquiterpènes/pharmacologie , Zanthoxylum/composition chimique
8.
Phytomedicine ; 102: 154192, 2022 Jul 20.
Article de Anglais | MEDLINE | ID: mdl-35636179

RÉSUMÉ

BACKGROUND: Nasopharyngeal carcinoma (NPC) is a type of malignant squamous cell tumour originating from the nasopharynx epithelium. Pentagalloylglucose (PGG) is a natural polyphenolic compound that exerts anticancer effects in many types of tumours. However, the role and underlying mechanism of PGG in NPC cells have not been fully defined. PURPOSE: This study aimed to investigate the anticancer activity of PGG as well as the potential mechanism in NPC cells. METHODS: The effects of PGG on the proliferation, apoptosis and cell cycle distribution of CNE1 and CNE2 cells were assessed by MTT and flow cytometry assays. Cell migration was evaluated using wound healing and transwell assays. The expression of microtubule-associated protein 1 light chain 3 beta (LC3B) was observed by immunofluorescence staining. Western blotting was used to explore the levels of related proteins and signalling pathway components. Furthermore, the effects of PGG on NPC cell growth were analysed in a xenograft mouse model in vivo using cisplatin as a positive control. RESULTS: PGG dose-dependently inhibited the proliferation of CNE1 and CNE2 cells. PGG regulated the cell cycle by altering p53, cyclin D1, CDK2, and cyclin E1 protein levels. PGG induced apoptosis and autophagy in NPC cells and elevated the Bax/Bcl-2 ratio and the protein levels of LC3B. Moreover, PGG decreased NPC cell migration by increasing E-cadherin and decreasing N-cadherin, vimentin and CD44 protein levels. Mechanistically, PGG treatment downregulated p-mTOR and ß-catenin expression but upregulated p-p38 MAPK and p-GSK3ß expression. In addition, PGG significantly inhibited NPC cell tumour growth and lung metastasis in vivo. CONCLUSION: PGG may suppress cell proliferation, induce apoptosis and autophagy, and decrease the metastatic capacity of NPC cells through the p38 MAPK/mTOR and Wnt/ß-catenin pathways. The present study provides evidence for PGG as a potential therapy for NPC.


Sujet(s)
Tanins hydrolysables , Cancer du nasopharynx , Tumeurs du rhinopharynx , Animaux , Apoptose/effets des médicaments et des substances chimiques , Lignée cellulaire tumorale , Mouvement cellulaire/effets des médicaments et des substances chimiques , Prolifération cellulaire/effets des médicaments et des substances chimiques , Glycogen synthase kinase 3 beta/métabolisme , Humains , Tanins hydrolysables/pharmacologie , Souris , Cancer du nasopharynx/traitement médicamenteux , Cancer du nasopharynx/métabolisme , Cancer du nasopharynx/anatomopathologie , Tumeurs du rhinopharynx/traitement médicamenteux , Tumeurs du rhinopharynx/métabolisme , Tumeurs du rhinopharynx/anatomopathologie , Sérine-thréonine kinases TOR/métabolisme , bêta-Caténine/métabolisme , p38 Mitogen-Activated Protein Kinases/métabolisme
9.
Fitoterapia ; 153: 104990, 2021 Sep.
Article de Anglais | MEDLINE | ID: mdl-34246746

RÉSUMÉ

Three novel lignans (1, 5 and 6) and two novel quinic acids (16 and 17) along with 15 known phenylpropanoids were obtained from the ethanol extract of Zanthoxylum nitidum var. tomentosum (Rutaceae). Their structures were confirmed by comprehensive spectroscopic data (NMR and HRESIMS), and the absolute configurations of all novel compounds were elucidated based on electronic circular dichroism (ECD) spectroscopic data. The production of nitric oxide (NO) in BV-2 microglial cells induced through lipopolysaccharide (LPS) was used to evaluate in vitro anti-neuroinflammatory activity of compounds 1-20. Compound 2, 3, 7 and 16 showed excellent inhibition of LPS-induced NO production. The structure-activity relationships of the isolates were investigated. In addition, the mechanism of action of 2 was elucidated by RT-PCR and Western blotting analysis, which indicated that it reduced neuroinflammatory mainly through NLRP3/caspase1 signaling pathways in LPS-induced BV2 microglial cells.


Sujet(s)
Anti-inflammatoires/pharmacologie , Lignanes/pharmacologie , Microglie/effets des médicaments et des substances chimiques , Zanthoxylum/composition chimique , Animaux , Anti-inflammatoires/isolement et purification , Lignée cellulaire , Chine , Lignanes/isolement et purification , Souris , Structure moléculaire , Monoxyde d'azote/métabolisme , Composés phytochimiques/isolement et purification , Composés phytochimiques/pharmacologie , Transduction du signal/effets des médicaments et des substances chimiques
10.
Medchemcomm ; 10(4): 584-597, 2019 Apr 01.
Article de Anglais | MEDLINE | ID: mdl-31057738

RÉSUMÉ

A series of asiatic acid (AA) based 1,2,3-triazole derivatives were designed, synthesized and subjected to a cell-based NF-κB inhibition screening assay. Among the tested compounds, compound 6k displayed impressive NF-κB inhibitory activity with an IC50 value in the low micromolar range. A molecular docking study was performed to reveal key interactions between 6k and NF-κB in which the 1,2,3-triazole moiety and the hydroxyl groups of the AA skeleton were important for improving the inhibitory activity. Subsequently, surface plasmon resonance analysis validated the high affinity between compound 6k and NF-κB protein with an equilibrium dissociation constant (KD) value of 0.36 µM. Further studies showed that compound 6k observably inhibited the NF-κB DNA binding, nuclear translocation and IκBα phosphorylation. Moreover, in vitro antitumor activity screening showed that compound 6k (IC50 = 2.67 ± 0.06 µM) exhibited the best anticancer activity against A549 cells, at least partly, by inhibition of the activity of NF-κB. Additionally, the treatment of A549 cells with compound 6k resulted in apoptosis induction potency and in vitro cell migration inhibition. Thus, we conclude that AA based 1,2,3-triazole derivatives may be potential NF-κB inhibitors with the ability to induce apoptosis and suppress cell migration.

11.
Article de Chinois | WPRIM (Pacifique Occidental) | ID: wpr-855538

RÉSUMÉ

Objective: To explore the correlation between the hemostatic activity and commercial grades of Panax notoginseng. Methods: After treatment with different commercial grades of P. notoginseng, the mice blooding time (BT) and clotting time (CT) were measured. At the same time the prothrombin time (PT), activation part thrombin time (APTT), fibrinogen (FIB), and platelet count (PLT) were also detected by fully automated blood cell analyzer. The contents of dencichine were determined by HPLC. Results: Mice BT, CT, PT, and APTT were decreased and PLT was increased significantly in all the commercial grades of P. notoginseng. Among all the commercial grades, the countless Tou Sanqi Powder has the best hemostatic activity. There is no correlation between the commercial grades and homostatic activity. Different contents of the extracted dencichine were observed among different commercial grades of P. notoginseng. The highest content of dencichine was 0.98% in countless Tou Sanqi and the lowest was 0.52% in 80 Tou Sanqi. There is no statistical correlation between the dencichine and hemostatic activity or dencichine and commercial grades of P. notoginseng. Conclusion: There is no statistical correlation between the commercial grades of P. notoginseng and haemostatic activity or dencichine contents. The dencichine in countless Tou Sanqi which is the cheapest of all displays the better hemostaic activity than those in 13 or 20 Tou Sanqi whose cost is almost as ten times as countless Tou Sanqi's.

12.
Article de Anglais | WPRIM (Pacifique Occidental) | ID: wpr-344995

RÉSUMÉ

<p><b>OBJECTIVE</b>To explore the effects of Zuojin Pills and its similar formulas on the stomach cold syndrome in a Wei cold model in rats.</p><p><b>METHODS</b>The rat Wei cold model was established by intragastric administration of glacial NaOH, and the gastric mucosa injury indices, together with the levels of motilin and gastrin in the stomach, were determined. The preventive and curative effects of Zuojin Pills and its similar formulas on gastric mucosa injury were investigated.</p><p><b>RESULTS</b>Zuojin Pills and its similar formulas could protect the gastric mucosa in the gastric cold model in rats at different levels. Fanzuojin Pills had the best effect in inhibiting gastric mucosa injury.</p><p><b>CONCLUSION</b>The different pharmacological effects of Zuojin Pills and its similar formulas in the rat gastric cold model were partially correlated with the degrees in cold and heat properties of the formulas.</p>


Sujet(s)
Animaux , Rats , Chimie pharmaceutique , Basse température , Modèles animaux de maladie humaine , Évaluation préclinique de médicament , Médicaments issus de plantes chinoises , Chimie , Pharmacocinétique , Pharmacologie , Muqueuse gastrique , Anatomopathologie , Malonaldéhyde , Sang , Rat Sprague-Dawley , Maladies de l'estomac , Sang , Traitement médicamenteux , Anatomopathologie , Superoxide dismutase , Sang , Comprimés , Équivalence thérapeutique
13.
Article de Anglais | WPRIM (Pacifique Occidental) | ID: wpr-236265

RÉSUMÉ

<p><b>OBJECTIVE</b>To investigate the effect of Radix Isatidis polysaccharides (RIP) on the immunological function and expression of immune related cytokines in mice.</p><p><b>METHODS</b>Sixty mice were randomly divided into six groups, the normal group, cyclophosphamide (Cy) model group, Levamisole (positive control) group, RIP low, medium and high dose groups (0.08 g/kg, 0.16 g/kg, 0.32 g/kg, respectively), ten in each group. By detecting the value of abdominal macrophage phagocytic index, serum hemolysin (HC50), proliferation of lymphocytes and expression of related cytokines, interleukin (IL-2) and interferon gamma (INF-gamma), the effect of RIP on immunological function and its mechanism were studied.</p><p><b>RESULTS</b>RIP could improve the level of hemolysin in immunological function depressed mice. The results showed that the value of macrophage phagocytic index in the high dose RIP group increased from 1.11+/-0.13 to 1.42+/-0.26. The level of IL-2 and INF-gamma could be decreased by Cy to 38.12+/-6.88 ng/L and 139.23+/-29.87 ng/L, respectively, while RIP in high dose could increase the secretion of IL-2 and INF-gamma to 53.54+/-14.43 ng/L and 189.91+/-32.63 ng/L, respectively.</p><p><b>CONCLUSION</b>RIP could enhance non-specific immunological function, humoral immunity and cellular immunity in mice.</p>


Sujet(s)
Animaux , Souris , Prolifération cellulaire , Poulets , Cyclophosphamide , Pharmacologie , Cytokines , Sang , Métabolisme , Médicaments issus de plantes chinoises , Pharmacologie , Immunité , Immunosuppression thérapeutique , Interféron gamma , Sang , Interleukine-2 , Sang , Macrophages , Biologie cellulaire , Souris de lignée BALB C , Phagocytose , Polyosides , Pharmacologie , Rate , Biologie cellulaire
14.
Acta Pharmacol Sin ; 28(11): 1761-74, 2007 Nov.
Article de Anglais | MEDLINE | ID: mdl-17959027

RÉSUMÉ

AIM: To investigate the effects of the wingless-related MMTV integration site 3A (Wnt3a) signaling on the proliferation, migration, and the myogenic and adipogenic differentiation of rat bone marrow mesenchymal stem cells (rMSC). METHODS: Primary MSC were isolated and cultured from Sprague-Dawley rats and characterized by flow cytometry. Mouse L cells were transfected with Wnt3a cDNA, and conditioned media containing active Wnt3a proteins were prepared. Cell proliferation was evaluated by cell count and 5-bromodeoxyuridine incorporation assay. The migration of rMSC was performed by using a transwell migration and wound healing assay. The myogenic and adipogenic differentiation in rMSC were examined by light microscopy, immunofluorescence, and RT-PCR at different time points after myogenic or adipogenic introduction. RESULTS: Wnt3a signaling induced beta-catenin nuclear translocation and activated the Wnt pathway in rMSC. In the presence of Wnt3a, rMSC proliferated more rapidly than the control cells, keeping their differentiation potential. Moreover, Wnt3a signaling induced 2.62% and 3.76% of rMSC-expressed desmin and myosin heavy chain after being cultured in myogenic medium. The myogenic differentiation genes, including Pax7, MyoD, Myf5, Myf4, and myogenin, were activated after Wnt3a treatment. On the other hand, Wnt3a inhibited the adipogenic differentiation in rMSC through the downregulated expression of CCAAT/enhancer-binding protein alpha (C/EBPalpha) and peroxisome proliferator-activated receptor gamma (PPARgamma). Furthermore, Wnt3a promoted the migration capacity of rMSC. CONCLUSION: The results indicate that Wnt3a signaling can induce myogenic differentiation in rMSC. Wnt3a signaling is also involved in the regulation of the proliferation and migration of rMSC. These results could provide a rational foundation for cell-based tissue repair in humans.


Sujet(s)
Cellules de la moelle osseuse/cytologie , Cellules de la moelle osseuse/effets des médicaments et des substances chimiques , Différenciation cellulaire/effets des médicaments et des substances chimiques , Mouvement cellulaire/effets des médicaments et des substances chimiques , Prolifération cellulaire/effets des médicaments et des substances chimiques , Cellules souches mésenchymateuses/cytologie , Cellules souches mésenchymateuses/effets des médicaments et des substances chimiques , Développement musculaire/effets des médicaments et des substances chimiques , Protéines de type Wingless/pharmacologie , Adipogenèse/effets des médicaments et des substances chimiques , Animaux , Techniques in vitro , Cellules L (lignée cellulaire) , Mâle , Souris , Rats , Rat Sprague-Dawley , Transduction du signal , Protéines de type Wingless/administration et posologie , Protéines de type Wingless/génétique , Protéine Wnt3 , Protéine Wnt3A , bêta-Caténine/métabolisme
15.
Sheng Wu Gong Cheng Xue Bao ; 23(1): 27-32, 2007 Jan.
Article de Chinois | MEDLINE | ID: mdl-17366884

RÉSUMÉ

Construction of recombinant adenovirus, which contain human microdystrophin, and then transfection into mesenchymal cells( MSCs) of mdx mice were done, and genetically-corrected isogenic MSCs were acquired; the MSCs transplantation into the mdx mice was then done to treat the Duchenne muscular dystrophy( DMD). Microdystrophin cDNA was obtained from recombinant plasmid pBSK-MICRO digested with restrictive endonuclease Not I ; the production was inserted directionally into pShuttle-CMV. The plasmid of pShuttle-CMV-MICRO was digested by Pme I , the fragment containing microdystrophin was reclaimed and transfected into E. coli BJ5183 with plasmid pAdeasy-1. After screening by selected media, the extracted plasmid of positive bacteria was transfected into HEK293 cells with liposome and was identified by observing the CPE of cells and by the PCR method. Finally, MSCs of mdx mice were infected with the culture media containing recombinant adenovirus, and the expression of microdystrophin was detected by RT-PCR and immunocytochemistry. Recombinant adenovirus including microdystrophin was constructed successfully and the titer of recombinant adenovirus was about 5.58 x 10(12) vp/mL. The recombinant adenovirus could infect MSC of mdx mice and microdystrophin could be expressed in the MSC of mdx mice. Recombinant adenovirus including microdystrophin was constructed successfully, and the microdystrophin was expressed in the MSC of mdx mice. This lays the foundation for the further study of microdystrophin as a target gene to correct the dystrophin-defected MSC for stem cell transplantation to cure DMD.


Sujet(s)
Adenoviridae/génétique , Dystrophine/génétique , Cellules souches mésenchymateuses/métabolisme , Protéines de fusion recombinantes/génétique , Animaux , Cellules cultivées , Dystrophine/métabolisme , Expression des gènes , Thérapie génétique/méthodes , Vecteurs génétiques/génétique , Humains , Immunohistochimie , Transplantation de cellules souches mésenchymateuses/méthodes , Cellules souches mésenchymateuses/cytologie , Souris , Souris de lignée mdx , Myopathie de Duchenne/génétique , Myopathie de Duchenne/anatomopathologie , Myopathie de Duchenne/thérapie , Protéines de fusion recombinantes/métabolisme , RT-PCR , Transduction génétique
16.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 29(4): 543-7, 2007 Aug.
Article de Chinois | MEDLINE | ID: mdl-19209802

RÉSUMÉ

OBJECTIVE: To investigate the clinical and lab features of sibling brother and sister both with Duchenne muscular dystrophy (DMD). METHODS: We conducted comprehensive clinical and lab investigations including the test of serum enzymes, electromyography (EMG), electrocardiography, color Doppler echocardiography, HE staining of skeletal muscles, immunohistochemical study of dystrophin and utrophin, multiple ligation probe amplification (MLPA) on exon 1-79 of dystrophin gene, and short tandem repeat-poly- merase chain reaction of CA repeats located in dystrophin gene. RESULTS: These two patients were confirmed to suffer from DMD. They were characterized by typical features of DMD including typical clinical manifestations, increased serum enzymes, EMG presenting myogenic impairment, HE staining presentation belonging to DMD, negative dystrophin in brother, and inconstantly positive on the sarcolemma of sister. Furthermore, no deletion or duplication was found in the 1-79 exons of dystrophin gene. The suffering brother and sister carried the same maternal X chromosome. CONCLUSIONS: Carriers of DMD gene show typical clinical and laboratory manifestations of DMD. Comprehensive examinations should be performed for such carriers.


Sujet(s)
Myopathie de Duchenne , Dystrophine/génétique , Femelle , Liaison génétique , Hétérozygote , Humains , Mâle , Myopathie de Duchenne/génétique , Myopathie de Duchenne/métabolisme , Myopathie de Duchenne/physiopathologie , Fratrie
17.
Nan Fang Yi Ke Da Xue Xue Bao ; 26(3): 261-5, 2006 Mar.
Article de Chinois | MEDLINE | ID: mdl-16546722

RÉSUMÉ

OBJECTIVE: To construct the eukaryotic expression vector of human microdystrophin gene and observe its expression in rat mesenchymal stem cells (rMSCs) in vitro. METHODS: The plasmid PBSK-MICRO containing human microdystrophin cDNA was digested by restriction endonuclease, and the resultant microdystrophin fragment was inserted into the NotI site of pcDNA3.1(+) to prepare the eukaryotic expression vector-pcDNA3.1(+)/ microdystrophin, which was identified by endonuclease digestion and sequencing. The recombinant plasmid was transfected into rMSCs via lipofectamine, and after G418 selection, the expression of microdystrophin was detected by RT-PCR and indirect immunofluorescence assay. RESULTS: Microdystrophin gene fragment was correctly inserted into the plasmid pcDNA3.1(+), as conformed by sequencing and digestion with Not I and Hind III. The total mRNA of the transfected rMSCs was extracted and microdystrophin mRNA expression was found in the cells by RT-PCR. Indirect immunofluorescence assay for the protein expression of microdystrophin showed bright red fluorescence in the transfected rMSCs. CONCLUSION: Eukaryotic expression plasmid pcDNA3.1(+)/microdystrophin has been constructed successfully and microdystrophin can be expressed in transfected rMSCs in vitro, which may facilitate further research of Duchenne muscular dystrophy treatment by genetically modified allogeneic stem cell transplantation.


Sujet(s)
Dystrophine/génétique , Cellules souches mésenchymateuses/métabolisme , Fragments peptidiques/génétique , Animaux , Séquence nucléotidique , Cellules cultivées , Dystrophine/biosynthèse , Technique d'immunofluorescence indirecte , Expression des gènes , Humains , Cellules souches mésenchymateuses/cytologie , Données de séquences moléculaires , Fragments peptidiques/biosynthèse , Plasmides/génétique , ARN messager/biosynthèse , ARN messager/génétique , ARN messager/métabolisme , Rats , Rat Sprague-Dawley , RT-PCR , Transfection
18.
Nan Fang Yi Ke Da Xue Xue Bao ; 26(1): 53-8, 2006 Jan.
Article de Chinois | MEDLINE | ID: mdl-16495176

RÉSUMÉ

OBJECTIVE: To investigate the effect of bone marrow stem cell transplantation (BMT) on the diaphragm muscles of mdx mice, a mouse model of Duchenne muscular dystrophy (DMD). METHODS: The bone marrow-derived stem cells form male SD rats was transplanted through the tail vein into 18 female 8-week-old mdx mice, which were sacrificed at 4, 8 and 12 weeks after BMT (6 at each time point), respectively. The diaphragm muscles of the mice were subjected to HE staining, immunofluorescence detection of dystrophin, reverse transcription (RT)-PCR analysis of dystrophin mRNA transcripts and PCR analysis of Sry (sex-determining region on the Y chromosome) gene, with age-matched female C57 mice and untreated mdx mice as the controls. RESULTS: The proportion of centrally nucleated fibers (CNF) in the diaphragm muscle of the recipient mdx mice was (15.58+/-0.91) %, (12.50+/-1.87) % and (10.17+/-1.17) % at 4, 8 and 12 weeks after BMT, respectively, significantly smaller than that of untreated mdx mice [(19.5+/-1.87) %], and the fibers after BMT showed less inflammatory infiltration. Compared with the untreated mice, the recipient mdx mice showed green fluorescence on significantly more diaphragm muscle cell membranes [with the proportion of dystrophin-positive fibers of (1.00+/-0.32) %, (6.00+/-1.05) % and (11.92+/-1.11) % at 4, 8, and 12 weeks after BMT]. RT-PCR of dystrophin mRNA also demonstrated significantly higher relative levels of dystrophin in the recipient mdx mice (0.19+/-0.05, 0.26+/-0.06 and 0.36+/-0.04 at 4, 8 and 12 weeks after BMT) than in untreated mdx mice, and Sry gene was present in the recipient mice. CONCLUSION: BMT can partially restore dystrophin expression and ameliorate the pathology in the diaphragm muscles of mdx mice, and has great potential to produce general therapeutic effect in patients with DMD.


Sujet(s)
Transplantation de moelle osseuse , Muscle diaphragme/métabolisme , Dystrophine/biosynthèse , Myopathie de Duchenne/chirurgie , Animaux , Transplantation de moelle osseuse/méthodes , Muscle diaphragme/anatomopathologie , Dystrophine/génétique , Femelle , Mâle , Souris , Souris de lignée C57BL , Souris de lignée mdx , Myopathie de Duchenne/métabolisme , Myopathie de Duchenne/anatomopathologie , Rats , Rat Sprague-Dawley , Transplantation hétérologue
19.
Article de Anglais | WPRIM (Pacifique Occidental) | ID: wpr-314149

RÉSUMÉ

<p><b>OBJECTIVE</b>To explore the activities of Composite Artemisia Capillaris Tablet (CACT) against hepatitis B virus replication in vitro.</p><p><b>METHODS</b>By means of radioimmunoassay (RIA), Dot blot and Southern blot, the surface and e antigen production of 2.2.15 cells, HBV DNA in 2.2.15 cell culture medium and that in 2.2.15 cells were examined respectively.</p><p><b>RESULTS</b>HBsAg, HBeAg values of 2.2.15 cells treated by CACT were lower than those of the control, the HBV DNA quantities in culture medium and in 2.2.15 cells decreased as compared with those cells with no treatment by CACT given to them.</p><p><b>CONCLUSION</b>CACT could inhibit HBV DNA replication, showing its potential antiviral activity in hepatitis B treatment.</p>


Sujet(s)
Humains , Lignée cellulaire tumorale , ADN viral , Antigènes de surface du virus de l'hépatite B , Métabolisme , Antigènes e du virus de l'hépatite virale B , Métabolisme , Virus de l'hépatite B , Génétique , Allergie et immunologie , Physiologie , Médecine traditionnelle chinoise , Préparations à base de plantes , Pharmacologie , Toxicité , Dosage radioimmunologique , Comprimés , Réplication virale
20.
Article de Chinois | WPRIM (Pacifique Occidental) | ID: wpr-282219

RÉSUMÉ

By referred to a lot of data, some new drug delivery systems(DDSs) including the Sustained and Controlled DDS, the Targeted DDS, the Transdermal DDS, the Bioadhensive DDS, the PowderJect DDS and the Self-Emulsifying DDS and their applications in TCD since 2000, will be summarized and some latest DDSs in the world including drug-eluting stents, gene therapy carrier system, biological chip, biomolecular motor-powered nanodevice and nanotrap will be also introduced in this paper. The objective of this paper is to introduce the new DDSs proceedings of and their applications in the Traditional Chinese Drugs(TCDs) and to provide some references for the pharmaceutics of TCD. For several recent years, the great success have been achieved in studying the new DDS application in the change of preparation of TCD by the investigators at home, but there is a large difference between at home and at board. So it is necessary to make a greater advance. During the modernization of TCD, there is an effective way that the new drug delivery systems(DDSs) will be applied in the change of the preparation of TCD.


Sujet(s)
Animaux , Humains , Administration par voie cutanée , Préparations à action retardée , Voies d'administration de substances chimiques et des médicaments , Vecteurs de médicaments , Systèmes de délivrance de médicaments , Médicaments issus de plantes chinoises , Nanotechnologie , Plantes médicinales , Chimie , Absorption cutanée , Technologie pharmaceutique
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