Bufonis Venenum,the dried secretion of Bufo bufo gargarizans or B. melanostictus,is toxic and hard with the efficacy of removing toxicity for detumescence and relieving pain. The processing of Bufonis Venenum dates back to the Song dynasty. In addition to the wine-processing,milk-processing and talcum powder-processing,there were some other kinds of processing methods in ancient times,such as baking,calcining,water-soaking and vinegar-processing. Modern studies have shown that the Bufonis Venenum has the main chemical components of bufadienolides,indole alkaloids sterols,and other compounds. It has the pharmacological effects of antitumor,cardiac,antibacterial,and analgesic activities,local anesthesia,and so on. This paper reviews the processing evolution,chemical components and pharmacological effects of Bufonis Venenum,providing references for its special processing and modern research as well as the theoretical basis for the research on its processing mechanism and quality control.
Animals ; Bufanolides/pharmacology* ; Bufonidae ; Quality Control

OBJECTIVETo observe effects of cold- and hot-property herbs and effects of hot and cold constitutions on the tumor growth of tumor bearing rats, and to observe the effect of Cinobufacini Injection (CI) on the tumor growth of tumor bearing rats of different constitutions.

METHODSEighty healthy male Wistar rats were randomly divided into eight groups, i.e., the tumor bearing control group, the tumor bearing heat syndrome group, the tumor bearing cold syndrome group, the heat syndrome tumor bearing group, the cold syndrome tumor bearing group, the tumor bearing CI group, the heat syndrome tumor bearing CI group, and the cold syndrome tumor bearing CI group, respectively. The weight and volume of rats' subcutaneous tumor were measured 14 days after tumor inoculation.

RESULTSThe weight and volume of tumor in the heat syndrome tumor bearing CI group [(3.55 +/- 1.12) g, (2864.44 +/- 1430.51) mm3] and the tumor bearing CI group [(4.29 +/- 1.14) g, (3397.19 +/- 1701.13) mm3] were significantly lower than those of the tumor bearing control group [(6.01 +/- 2.45) g, (6218.91 +/- 3837.64) mm3] and the cold syndrome tumor bearing CI group [(6.90 +/- 1.57) g, (6168.42 +/- 2457.03) mm3], showing statistical difference (P<0.05). There was insignificant difference among other groups.

CONCLUSIONSCI showed better tumor inhibition effects on tumor bearing rats of heat syndrome constitution, which indicated CI was of cold property. It might be possibly used in tumor bearing rats of heat syndrome constitution.

Animals ; Bufanolides ; pharmacology ; therapeutic use ; Injections ; Male ; Neoplasms, Experimental ; drug therapy ; pathology ; Rats ; Rats, Wistar

Bufonis Venenum, an animal medicinal material, is widely used for treating cardiovascular diseases and pain induced by rheumatics or malignant tumors. In view of the high activity and high toxicity, it is of great significance to pay attention to the quality control of Bufonis Venenum to ensure the safety and effectiveness of its preparations. China's drug standards involve 102 preparations(474 batch numbers) containing Bufonis Venenum approved for sale, including 14 preparations in the Chinese Pharmacopoeia(2020 edition) and 68 preparations in the standards issued by the Ministry of Health Drug Standard of the People's Republic of China. Bufonis Venenum is mostly used in pill and powder preparations in the form of raw powder, with the main functions of clearing heat, removing toxin, relieving swelling and pain, replenishing qi, activating blood, opening orifice, and awakening brain. Except the high level of quality control for Bufonis Venenum in the preparations in the Chinese Pharmacopoeia(2020 edition), the quality control standards of Bufonis Venenum in other preparations are low or even absent. Therefore, it is urgent to conduct research on the improvement of quality standards for the preparations containing Bufonis Venenum. This study retrieved the reports focusing on the quality evaluation and quality control of the preparations containing Bufonis Venenum from CNKI, PubMed, and Web of Science. Qualitative and quantitative analysis methods for 64 preparations containing Bufonis Venenum have been reported, mainly including thin-layer chromatography, HPLC fingerprint, and multi-component content determination. The index components mainly involved bufadienolides, such as gamabufalin, arenobufagin, bufotalin, bufalin, cinobufagin, and resibufogenin. According to the literature information, this paper suggests that attention should be paid to the correlations between the analysis methods and detection indexes of medicinal materials, decoction pieces and preparations, the monitoring of indole alkaloids, and the content uniformity inspection for further improving the quality standards for the preparations containing Bufonis Venenum.
Animals ; Humans ; Bufonidae ; Powders ; Bufanolides/pharmacology* ; Quality Control ; Chromatography, High Pressure Liquid ; Pain/drug therapy*

Bufalin is an active compound of the traditional Chinese medicine Chansu, which exhibits significant anti-tumor activities in many solid tumors and leukemia cell lines. Bufalin could introduce apoptosis, reverse drug-resistance, and prevent migration and invasion of tumor cells. This paper reviewed the latest research progress of the in vitro and in vivo anti-tumor effect and mechanism of bufalin on a series of cancers, such as hepatocellular carcinoma, lung cancer, colon cancer, gastric cancer, leukemia, bladder cancer, and its formulation study is also summarized for the reference of its further study and application.
Animals ; Antineoplastic Agents ; chemistry ; pharmacology ; therapeutic use ; Bufanolides ; chemistry ; pharmacology ; therapeutic use ; Chemistry, Pharmaceutical ; methods ; Neoplasms ; drug therapy ; pathology

OBJECTIVE: Bufalin is an effective drug for the treatment of liver cancer. But its high toxicity, poor water-solubility, fast metabolism and short elimination half-life limit its use in tumor treatment. How to make the drug accumulate in the tumor and reduce side effects while maintaining its efficacy are urgent problems to be solved. The goal of this study is to solve these problems. METHODS: A copolymer with tunable poly-N-isopropylacrylamide and polylactic acid was designed and synthesized. The corresponding dual targeting immunomicelles (DTIs) loaded with bufalin (DTIs-BF) were synthesized by copolymer self-assembly in an aqueous solution. The size and structure of DTIs-BF were determined by ZetaSizer Nano-ZS and transmission electron microscopy. Then, its temperature sensitivity, serum stability, critical micelle concentration (CMC), entrapment efficiency (EE), drug release and non-cytotoxicity of blank block copolymer micelles (BCMs) were evaluated. Next, the effects of DTIs-BF on cellular uptake, cytotoxicity, and tumor cell inhibition were evaluated. Finally, the accumulation of DTIs-fluorescein isothiocyanate (FITC) and the in vivo anti-tumor effect were observed using an interactive video information system. RESULTS: DTIs-BF had a small size, spherical shape, good temperature sensitivity, high serum stability, low CMC, high EE, and slow drug release. The blank BCMs had very low cytotoxicity. Compared with free bufalin, the in vitro cellular internalization and cytotoxicity of DTIs-BF against SMMC-7721 cells were significantly enhanced, and the effects were obviously better at 40 °C than 37 °C. In addition, the therapeutic effect on SMMC-7721 cells was further enhanced by the programmed cell death specifically caused by bufalin. When DTIs-FITC were injected intravenously in BALB/c nude mice bearing liver cancer, the accumulation of FITC was significantly increased in tumors. CONCLUSION DTIs-BF is a potentially effective nano-formulation and has broad prospects in the clinical treatment of liver cancer.
Animals ; Antineoplastic Agents/pharmacology* ; Bufanolides ; Cell Line, Tumor ; Liver Neoplasms/drug therapy* ; Mice ; Mice, Inbred BALB C ; Mice, Nude

OBJECTIVETo investigate the mechanisms underlying the effect of Chansu Injection (CHS) in reversing multi-drug resistance (MDR) of HL60/ADM cells.

METHODSMTT assay was used to investigate the effect of CHS on adramycin (ADM) sensitivity of HL-60/ADM cells. Flow cytometry was used to observe the effect of CHS on the cell cycle of HL60/ADM cell. The expressions of NF-κB, MRP, GST-π, and iNOS were detected by immunocytochemistry.

RESULTSTreatment with CHS lowered the IC(50) of ADM in HL60/ADM cells from 34.1971 µmol/L to 17.4393 µmol/L, and caused an increase in G(0)/G1 and G(2)/M phase cells with decreased S phase cells. CHS decreased the expressions of MRP mRNA and GST-π and MRP proteins but increased the expressions of iNOS and NF-κB proteins in the cells.

CONCLUSIONCHS can partly reverse MDR in HL60/ADM cells possibly by down-regulating MRP and GST-π, up-regulating NF-κB and iNOS, and promoting cell apoptosis, thereby increase ADM sensitivity of HL-60/ADM cells.

Amphibian Venoms ; pharmacology ; Bufanolides ; pharmacology ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; HL-60 Cells ; Humans

OBJECTIVETo study the effect of cinobufagin (CBG) on HeLa cell proliferation, and to analyze its mechanism.

METHODSProliferation inhibition in vitro was evaluated by MTT and Sulforhodamine B (SRB) assays in several human tumor cell lines, including Bel-7402, HeLa, MCF-7, BGC-823 and HL60. The cycle of HeLa cells was analyzed by flow cytometry. Two-dimensional electrophoresis was applied to analyze the influence of CBG on HeLa cell proteomics.

RESULTSCBG had inhibitory effects on proliferation of five human cancer cell lines, and the IC(50) values were 0.011 micromol/L (Bel-7402), 0.019 micromol/L (HeLa), 0.116 micromol/L (MCF-7), 0.149 micromol/L (BGC-823) and 1.369 micromol/L (HL60), respectively. HeLa and Bel-7402 cells were among the most sensitive. Flow cytometry assay indicated that the treatment of HeLa cells with various concentrations of CBG for 72 h was able to increase the cell number at G(2)/M phase, from 17.3% up to 35.6%. The results of two-dimensional electrophoresis showed that treatment of HeLa cells with 0.02 micromol/L CBG for 48 h resulted in apparent changes of certain small molecular weight (30,000 - 90,000) acidic proteins (pH 4 - 6).

CONCLUSIONCinobufagin has significant inhibitory effect on growth of five human cancer cells in vitro. It may lead to cell cycle arrest of HeLa cells at G(2)/M phase. It can also change the expression of some small molecular acidic proteins in HeLa cells.

Antineoplastic Agents ; pharmacology ; Bufanolides ; pharmacology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; HeLa Cells ; Humans ; Materia Medica ; Pilot Projects

OBJECTIVETo examine the effect of cinobufacini on rat thoracic aorta and its mechanism.

METHODSIsolated rat thoracic aorta was perfused and isometric tension was recorded by organ bath technique before and after cinobufacini treatment.

RESULTCinobufacini induced contraction of isolated thoracic aorta with or without endothelium in a concentration-dependent manner (at concentration of 2.5,5.0,7.5,10.0 g/L). The vasoconstriction effect of cinobufacini was more potent in endothelium-denuded aorta ring [(16.3+/-3.39)%, (52.5+/-7.70)%, (60.9+/-8.84)%, (69.2+/-11.34)%] than in endothelium-intact aorta ring [(6.2+/-2.07)%, (14.7+/-4.91), (17.6+/-5.86)%, (20.3+/-6.78)% (P<0.01)]. Its contractile effect was attenuated in Ca(2+)-free solution (about 1/10 of that in buffer with 1.25 mmol/L CaCl(2)) or by the treatment with verapamil (10(-7)mol/L), an L-type calcium channel antagonist. Cinobufacini induced contraction on the endothelium-intact rat aorta was augmented by pretreatment with L-NAME (10(-4)mol/L), a nitric oxide synthase inhibitor.

CONCLUSIONCinobufacini contracts rat thoracic aorta by opening the voltage-dependent Ca(2+) channel and increasing Ca(2+) influx into vascular smooth muscle. Cinobufacini can also stimulate the release of vascular relaxant factor, nitric oxide, from the endothelium and thus antagonize cinobufacini-induced contraction.

Animals ; Aorta, Thoracic ; drug effects ; Bufanolides ; pharmacology ; Endothelium, Vascular ; drug effects ; metabolism ; In Vitro Techniques ; Male ; Nitric Oxide ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Vasoconstriction ; drug effects ; Vasoconstrictor Agents ; pharmacology

OBJECTIVETo study the bioaffinity between 8 bufadienolides(Bu) and tumor cells and analyze the correlation between the bioaffinity and the anti-tumor activities of Bu.

METHODMix and cultivate the chloroform extract of Chansu and MGC-803. Measure the content of 8 Bu in supernatant and cells using HPLC and calculate their affinity rate.

RESULTThe coefficient correlation between the decrease of Bu in cell supernatant after affinity and its MGC-803 restrictive activities, and between the cotent percentage of the free Bu in free cells with its MGC-803 restrictive activities, and between the difference between the decrease and the percentage and its MGC-803 restrictive activities is r = 0.82 (P < 0.05), r = -0.04 and r = 0.83 (P < 0.05) respectively.

CONCLUSIONEight Bu have different levels of affinity with MGC-803 which correlate with their anti-tumor activities.

Amphibian Venoms ; chemistry ; Animals ; Antineoplastic Agents ; isolation & purification ; pharmacology ; Anura ; Bufanolides ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Chromatography, High Pressure Liquid ; methods ; Neoplasms ; drug therapy

OBJECTIVETo seek possible effect targets of bufalin in HeLa cells by studying the impact of bufalin on cell protein expression profile after treatment on human cervical carcinoma cell lines HeLa.

METHODBufalin's ICs0was measured by MTr assay. The apoptosis of cells was observed by FCM (flow cytometry) and Hoechst 33342 staining assay. Differentiated expression protein spots were founded and identified using proteomic techniques, which could induce HeLa cell apoptosis.

RESULTBufalin showed remarkable cytotoxic effect on HeLa cells. IC50 (154 +/- 21.5) nmol X L(-1) indicated the possibility of inducing cell apoptosis. The protein expression profile showed 11 differentiated expression protein spots. Among the 11 proteins, nudix-type motif 5, vimentin, hnRNP C1/hnRNP C2 variant, HNRPK, HNRPK isoform a variant (two spots are the same protein), heat shock protein 27, macrophage-capping protein, SELENBP1 protein were down-regulated, while ribosomal protein, large, P0 and S-adenosylmethionine synthetase 2 were up-regulated by bufalin treatment. They may be effect targets of bufalin in HeLa cells. Western blotting showed consistent results in heat shock protein 27, vimentin and HNRPK between expression after treatment with bufalin and two-dimensional electrophoresis.

CONCLUSIONBufa-Lin can induce apoptosis in human cervical carcinoma cells HeLa and the effect of bufalin may be related to the joint intervention with multiple protein targets.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Cell Line, Tumor ; Female ; HeLa Cells ; Humans ; Neoplasm Proteins ; metabolism ; Proteomics ; methods ; Uterine Cervical Neoplasms ; drug therapy ; metabolism ; pathology