scholarly journals Gastrodia elata powder capsule enhances anti-epileptic effect of carbamazepine by decreasing P-gp expression

2021 ◽  
Vol 18 (9) ◽  
pp. 1859-1865
Author(s):  
Xiangji Dang ◽  
Pei Zhao ◽  
Yan Liu ◽  
Long Qin ◽  
Haisheng Jiao

Purpose: To investigate the influence of Gastrodia elata powder capsule (GC) or gastrodin (GTD) on the anti-epileptic effect of carbamazepine (CBZ) on penicillin (PG)-induced epilepsy in rats. Methods: A total 116 rats were used in this study. Rats in the control group (n = 8) were injected with normal saline (NS) in place PG. Epilepsy was induced in the remaining 108 rats on the first day via PG injection. The rats were then divided randomly into six groups (18 rats per group): PG group, CBZ group, CBZ + GC group, CBZ + GTD group, GC group, and GTD group, which were given (p.o.) NS, CBZ (100 mg/kg), CBZ (100 mg/kg.) + GC (350 mg/kg), CBZ (100 mg/kg) + GTD (100 mg/kg), GC (350 mg/kg), and GTD (100 mg/kg), respectively, once a day for 15 days. The behavioral characteristics of the rats were observed and used to assess the anti-epileptic effect of the test drugs. Real-time quantitative reverse transcription-PCR and Western blot assays were employed for the determination of the effect of CBZ, GC and GTD on the expression levels of P-gp. Results: CBZ significantly reduced the symptoms of epilepsy, while GC and GTD enhanced the antiepileptic effect of CBZ, and reversed the CBZ-induced increases in the protein expressions of mrd1a and P-gp (p < 0.05). Conclusion: GC reverses CBZ drug resistance, probably through downregulation of P-gp expression. This finding indicates that GC is a potential anti-epilepsy drug, but it merits further studies.

1999 ◽  
Vol 37 (12) ◽  
pp. 3879-3882 ◽  
Author(s):  
E. Falcone ◽  
M. Tarantino ◽  
L. Di Trani ◽  
P. Cordioli ◽  
A. Lavazza ◽  
...  

Determination of the G and P serotypes of group A bovine rotaviruses from 149 samples of feces or intestinal contents collected from calves showing clinical signs of neonatal diarrhea was performed by a nested reverse transcription-PCR typing assay. The G6 serotype was the most prevalent, accounting for viruses in 55.7% of the samples; viruses of the G10 and G8 serotypes were found in 34.9 and 4.7% of the samples, respectively. The virus in one sample (0.7%) was not classified due to concomitant infection with G6 and G8 strains, whereas viruses in six samples (4.0%) could not be characterized with any of the three G serotype-specific primers selected for the present study. When examined for their P-serotype specificities, viruses in 55 and 42.3% of the samples were characterized as P[11] and P[5], respectively, no P[1] serotype was identified, and viruses in 2.7% of the samples could not be classified due to multiple reactivity with both P[5]- and P[11]-specific primers. Various combinations of G and P serotypes were observed, the most frequent being G6,P[5] (38.3%), G10,P[11] (31.5%), and G6,P[11] (15.4%). The results of the present study, while contributing to a better understanding of the epidemiology of bovine rotaviruses in Italy, address the relevance of serotype specificity with regard to the constancy of the quality of bovine rotavirus vaccines under different field conditions.


Neurology ◽  
2018 ◽  
Vol 90 (24) ◽  
pp. e2146-e2154 ◽  
Author(s):  
Jay S. Charleston ◽  
Frederick J. Schnell ◽  
Johannes Dworzak ◽  
Cas Donoghue ◽  
Sarah Lewis ◽  
...  

ObjectiveTo describe the quantification of novel dystrophin production in patients with Duchenne muscular dystrophy (DMD) after long-term treatment with eteplirsen.MethodsClinical study 202 was an observational, open-label extension of the randomized, controlled study 201 assessing the safety and efficacy of eteplirsen in patients with DMD with a confirmed mutation in the DMD gene amenable to correction by skipping of exon 51. Patients received once-weekly IV doses of eteplirsen 30 or 50 mg/kg. Upper extremity muscle biopsy samples were collected at combined study week 180, blinded, and assessed for dystrophin-related content by Western blot, Bioquant software measurement of dystrophin-associated immunofluorescence intensity, and percent dystrophin-positive fibers (PDPF). Results were contrasted with matched untreated biopsies from patients with DMD. Reverse transcription PCR followed by Sanger sequencing of newly formed slice junctions was used to confirm the mechanism of action of eteplirsen.ResultsReverse transcription PCR analysis and sequencing of the newly formed splice junction confirmed that 100% of treated patients displayed the expected skipped exon 51 sequence. In treated patients vs untreated controls, Western blot analysis of dystrophin content demonstrated an 11.6-fold increase (p = 0.007), and PDPF analysis demonstrated a 7.4-fold increase (p < 0.001). The PDPF findings were confirmed in a re-examination of the sample (15.5-fold increase, p < 0.001). Dystrophin immunofluorescence intensity was 2.4-fold greater in treated patients than in untreated controls (p < 0.001).ConclusionTaken together, the 4 assays, each based on unique evaluation mechanisms, provided evidence of eteplirsen muscle cell penetration, exon skipping, and induction of novel dystrophin expression.Classification of evidenceThis study provides Class II evidence of the muscle cell penetration, exon skipping, and induction of novel dystrophin expression by eteplirsen, as confirmed by 4 assays.


2019 ◽  
Author(s):  
Zhiyong Wu ◽  
Xiaoyu Kong ◽  
Zhihui Wang

Abstract Background The aim was to investigate whether integrin α7 (ITGA7) influenced hepatocellular carcinoma (HCC) progression, and explore its effect on regulating endothelium-mesenchymal transformation (EMT).Methods ITGA7 mRNA and protein expressions in human normal liver epithelial cell line and HCC cell lines were determined by reverse transcription polymerase chain reaction (RT-qPCR) and western blot. ITGA7 siRNA (ITGA7-KD group) and nonsense siRNA (control group) were transfected into Huh7 cells and SUN449 cells. After transfection, ITGA7 mRNA and protein expressions (RT-qPCR and western blot), cell proliferation (Cell Counting Kit-8), apoptosis (Annexin V/Propidium Iodide assay), migration (Wound scratch assay) and invasion (Transwell assay) were determined. E-cadherin and α-SMA expressions (RT-qPCR and western blot) were determined.Results ITGA7 mRNA and protein expressions were increased in Li7, Huh7, SKHEP1 and SNU449 cells compared to THLE-3 cells. In both Huh7 and SNU449 cells, ITGA7 mRNA and protein expressions were decreased in ITGA7-KD group than control group after plasmids transfection, indicating the successful transfection. Then, cell proliferation was decreased at 48h and 72h; cell apoptosis rate was increased at 48h; cell migration rate was reduced at 24h; cell invasive count was decreased at 24h in ITGA7-KD group compared to control group. Furthermore, increased E-cadherin but decreased α-SMA mRNA and protein expressions were discovered in ITGA7-KD group than control group at 24h.Conclusions ITGA7 knockdown suppresses HCC progression and inhibits EMT process in HCC, indicating that ITGA7 might be a potential novel treatment target for HCC therapy.


2008 ◽  
Vol 46 (10) ◽  
pp. 3330-3337 ◽  
Author(s):  
Y.-P. Lin ◽  
C.-L. Kao ◽  
S.-Y. Chang ◽  
K. Taniguchi ◽  
P.-Y. Hung ◽  
...  

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 4406-4406 ◽  
Author(s):  
Yiqing Cai ◽  
Lili Feng ◽  
Dai Yuan ◽  
Qian Wang ◽  
Xin Wang

Abstract Introduction: CD39/CD73/ADO/A2A system plays important roles in tumor growth and therapy. Extracellular adenosine (ADO) receptor A2A (A2AR) is the dominant receptor expressed on chronic lymphocytic leukemia (CLL) cells. ADO has been shown to protect CLL cells from spontaneous and drug-induced apoptosis through A2AR activation. Overexpression of hypoxia inducible factor-1α (HIF-1α), an important mediator controlling the expression of a wide variety of apoptotic genes, has been observed in bone marrow leukemic cells from CLL patients. However, the reciprocal action of A2AR and HIF-1α in CLL remains elusive. This study was aimed to explore the molecular mechanisms underlying the relationship between A2AR activation and HIF-1α in CLL. Materials and Methods: Peripheral blood (PB) and bone marrow (BM) samples were collected from 30 healthy volunteers (control) and 20 patients who were diagnosed with CLL for the first time at the Hematology Department in our hospital. CD39/CD73/ADO/A2AR axis-related protein and HIF-1α protein expressions in CLL PB and BM tissues were examined by Western-blot. CD39, CD73, A2AR and HIF-1α expression on CLL cells were also determined by FACS. CLL cell line (Lym-2) was purchased from ATCC and incubated in DMEM medium supplemented with 10% FBS. The protein expressions of CD39, CD73 and A2AR on Lym-2 cells were confirmed by Western-blot. To study the impacts of A2AR activation and inactivation on CLL cells, CLL cells were treated with A2AR agonist CGS21680 or antagonist SCH58261, followed by determination of cell proliferation and HIF-1α protein expression. Results: The protein expressions of CD39, CD73, A2AR and HIF-1α were higher in CLL group than those in control group (BM: CD39 1.471 vs 0.926, CD73 1.097 vs 0.489, A2AR 1.139 vs 0.342 and HIF-1α 0.940 vs 0.362, P<0.05 Figure 1A; PB : CD39 1.809 vs 1.331, CD73 1.039 vs 0.653, A2AR 1.738 vs 1.119 and HIF-1α 1.336 vs 1.010, P<0.05 Figure 1B). Moreover, CD73 and HIF-1α protein expression was found to have relationship with disease stage. Patients who belonged to stage IV (Rai) and stage B/C (Binet) exhibited higher levels of CD73 and HIF-1α than patients who belonged to stage I to III (Rai) and stage A (Binet) . Data from FACS analysis showed that the proportion of CD39+CLL cells was higher than CD73+CLL cells and HIF-1α+CLL cells (P<0.05, Figure 2A). When compared to control group, the proportions of CD39+cells and CD73+ cells were significant higher in CLL group (Figure 2B). Western-blot results showed that Lym-2 cells expressed CD39, CD73 and A2AR simultaneously (Figure 3A). The results from CLL cells and A2AR agonist/antagonist incubation showed that when CGS21680 concentration was of ≥ 10uM, cell proliferation was promoted obviously (Figure 3B). On the contrary, SCH58261 could inhibit CLL cells proliferation when its concentration was of ≥5uM at a concentration dependent manner (Figure 3C). Moreover, HIF-1α protein expression was also affected by A2AR agonist at a concentration dependent manner(0uM 0.851, 10uM 1.116, 20uM 1.420, 50uM 1.41, Figure 3D). Conclusion: Our study showed that ADO produced by CD39 and CD73 could affect CLL cells proliferation and HIF-1α expression through A2AR-mediated mechanisms. The exploration of the molecular mechanisms underlying the relationship between A2AR activation and HIF-1α could help us to find a novel approach to CLL therapy. Disclosures: No relevant conflicts of interest to declare. Disclosures No relevant conflicts of interest to declare.


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