Neferine in the Lotus Plumule Potentiates the Antitumor Effect of Imatinib in Primary Chronic Myeloid Leukemia Cells In Vitro.

Imatinib, the prototype BCR-ABL tyrosine kinase inhibitor (TKI), is the first-line treatment for Philadelphia chromosome-positive chronic myeloid leukemia (CML) in the chronic phase. However, a subgroup of patients exhibit poor response or experience relapse. This issue may be overcome by combination therapy using natural compounds. Neferine, a major bisbenzylisoquinoline alkaloid extracted from "lotus plumule" (seed embryo of lotus) commonly used in traditional Chinese medicine and tea, was used herein in the combination treatment of CML. The MTT assay showed that neferine exerted cytotoxicity in primary CML cells in a dose-dependent manner. Moreover, low concentrations of neferine (4 and 8 µM) sensitized primary CML cells to imatinib (CI < 1), and significantly decreased its IC50 from 0.70 ± 0.10 to 0.32 ± 0.06 µM and 0.16 ± 0.02 µM, respectively. Cotreatment of neferine and imatinib significantly decreased the expression of BCR-ABL protein and its molecular chaperone heat shock protein 90 (Hsp90) mRNA and protein levels, and further decreased phospho-extracellular regulated protein kinase 1/2 (p-Erk1/2) and myeloid cell leukemia (Mcl-1) expression. These results suggest that neferine might be a potential imatinib sensitizer in CML treatment. PRACTICAL APPLICATION: In China, Lotus plumule, the green embryo of lotus, is used as a tea and as a source of herbal medicine in the treatment of anxiety, insomnia, spermatorrhea, and thirst. Additional, neferine, a bisbenzylisoquinoline alkaloid extracted from lotus plumule has been shown to have antitumor potential. Herein, the effect of neferine and imatinib cotreatment on primary CML cells obtained from CML patients was assessed, with a synergistic effect being observed between the two compounds. Therefore, neferine might be a promising natural compound to potentiate imatinib in CML patients.

Efficacy of oral fludarabine in patients with chronic lymphocytic leukemia/small lymphocytic lymphoma.

OBJECTIVE: To investigate the efficacy and safety of oral fludarabine in patients with chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL). METHODS: The patients received oral fludarabine 40 mg/(m2.d) for 5 consecutive days, each treatment lasting 4 weeks. The efficacy was assessed with National Comprehensive Cancer Network (NCCN) criteria for response. RESULTS: Twenty-two patients received the treatment, a median of 4 cycles per patient. The rate of complete response (CR), partial response (PR), and overall response (OR) was 40.9% (9/22), 45.5% (10/22), and 86.4% (19/22), respectively. Among the 17 previously untreated patients, 7 (41.2%) achieved CR and 8 (47.0%) achieved PR. Two of the 5 pre-treated patients achieved CR and the other 2 achieved PR. During a median observation of 24 months, the overall survival rate was 81.8%. The main adverse reactions were myelosuppression and infection. Grade 1 to 3 granulocytopenia was found in 7 (31.8%) patients, and infection in 3 (13.6%) patients. Nonhematologic toxicity was mild. All the adverse reactions were reversible. CONCLUSION: The oral fludarabine is effective, safe, and well-tolerated in the patients with CLL/ SLL.

[Effect of neferine combined with mdr-1shRNA on the expression of mdr-1/P-gp in K562/A02 cell line].

OBJECTIVE: To determine the effect of neferine (Nef) combined with mdr-1shRNA on the expression of mdr/P-gp in K562/A02 cell line. METHODS: MTT assay was used to observe the cell proliferation. The expression level of P-gp was determined by Western blot and the transcription of mdr-1 gene was detected by semi-quantitative RT-PCR. RESULTS: After K562/A02 cells were treated by Nef or mdr-1shRNA alone or both for 24 h, the proliferation of K562/A02 cells was significantly higher in the Nef combined with mdr-1shRNA treatment group than that of Nef or mdr-1shRNA alone group (P<0.01).The expression of mdr-1/P-gp in the Nef with mdr-1 shRNA group was significantly lower than that of Nef or mdr-1shRNA alone group. CONCLUSION: Nef enhances the inhibition of mdr-1shRNA expression vector on K562/A02 cell proliferation and on P-gp protein to effectively reverse multidrug resistance induced by mdr-1 gene encoding P-gp.

[Influence of neferine and erythromycin on cellular GSH concentration in K562/A02 cell line].

OBJECTIVE: To investigate the reversal effect of multidrug resistance (MDR) mediated by glutathione (GSH) detoxifcation system in K562/A02 cell line by neferine and erythromycin. METHODS: Cellular GSH concentration was examined by biochemical analyses with DTNB. RESULTS: Cellular GSH concentration in K562/A02 cell line [(137.34 +/- 33.10) mg/mgprot] was higher than that in K562 cell line [(73.38+/-10.02) mg/mgprot] (P < 0.05), and 1.87-fold greater in K562/A02 cell line compared with that in K562 cell line. Cellular GSH concentration in K562/A02 cell line treated with Nef10 mg/L, EM100 mg/L, Nef10 mg/L + EM100 mg/L, and VRP5 mg/L for 2 days [(69.01 +/- 15.99) mg/mgprot, (70.57 +/- 11.93) mg/mgprot, (65.74 +/- 13.37) mg/mgprot, and (70.86 +/- 16.16) mg/mgprot, respectively] decreased than that in K562/A02 cell line without drugs (P < 0. 05). CONCLUSION: Increase of cellular GSH concentration was one of the MDR mechanisms in K562/A02 cell line. Decrease of cellular GSH concentration may be one of the MDR reversal mechanisms in K562/A02 cell line by neferine and erythromycin.

[Antibacterial activity of Chinese medicine kou-kang mouthwash and its acute toxicity test].

OBJECTIVE: To study the antibacterial activity of Chinese medicine kou-Kang mouthwash and its acute toxicity, and provide the basis of experiment and therapy for infectious diseases of the oral cavity. METHODS: Minimal inhibitory concentrations (MIC) of kou-Kang mouthwash were determined by the agar dilution method, and the acute toxicity was done. RESULTS: The range of MIC kou-Kang mouthwash against 27 Gram negative bacterial were 0.03-1.0 g.ml-1. MIC50 was 0.125 g.ml-1 and MIC90 was 0.125-0.5 g.ml-1. The range of MIC kou-Kang mouthwash against 21 Gram positive bacterial was 0.008-0.5 g.ml-1. The range of MIC50 and MIC90 were 0.015-0.125 g.ml-1 and 0.015-0.5 g.ml-1, respectively. The value of MIC was 0.015 g.ml-1 against candidia albicans. The acute toxic test indicated that the maximum tolerance dose was 417.6 g.kg-1 and 210 times as much as the clinical dose a day. No death of animals or toxicity and side-effect occurred in the test. CONCLUSION: Kou-Kang mouthwash shows a potent antibacterial activity against various pathogens of the oral cavity. Its antibiotic activity against Gram positive bacteria was 2-8 times stronger than that against Gram negative bacteria. It has low toxicity and can be used in clinical application.