In vitro models of human colon carcinoma cell line(Caco-2 cell monolayer) and human intestinal bacteria were used to investigate the intestinal transport and biotransformation of resibufogenin and cinobufagin in Chan ...In vitro models of human colon carcinoma cell line(Caco-2 cell monolayer) and human intestinal bacteria were used to investigate the intestinal transport and biotransformation of resibufogenin and cinobufagin in Chan Su by HPLC/APCI-MSn. The experimental results of Caco-2 cell monolayer demonstrate that the apparent permeability coefficients(Papp) of resibufogenin and cinobufagin are higher than 10–6 cm/s, which indicates that both resibufogenin and cinobufagin have a good absorption in the small intestine. And the biotransformation result of human intestinal bacteria shows that resibufogenin has been transformed to 3-epiresibufogenin and cinobufagin has been transformed to 3-epicinobufagin, deacetylcinobufagin and 3-epideacetycinobufagin, respectively.展开更多
An endogenous inhibitor of the sodium pump from the toad venom Chan′su was purified. The dry substance Chan′su was extracted with methanol. The dry residue was passed subsequently through membrane filters with the e...An endogenous inhibitor of the sodium pump from the toad venom Chan′su was purified. The dry substance Chan′su was extracted with methanol. The dry residue was passed subsequently through membrane filters with the exclusion size of 10 000, 3 000 and 1 000 at 1 psi in a Filtron Pro Vario-3-System and applied to thin layer chromatography made of Silica 60 F 254+366 separated in CHCl 3∶MeOH∶H 2O. The R f 0.55 fractions inhibiting the sodium pump were purified on a HPLC-RP C18 using a linear H\-2O/methanol gradient at 220 nm and 250 nm on DAD detector. The bio-activity was measured by a specific enzyme-linked immunosorbent assay (ELISA) and 86 Rb-uptake into human red blood cells. In Chan′su there exists an inhibitor of the sodium pump which interacts with antibodies against ouabain and proscillaridin A, and inhibits the sodium pump activity in the red blood cells. The results show that a low molecular weight compound has been isolated from the toad venom Chan′su. It inhibits the sodium pump, interacts preferentially with antibodies against ouabain. It is water soluble and shows a maximum of absorbance at 250 nm. Several milligrams of the compounds from one kilogram were obtained in pure form. The inhibitor differs considerably in its chemical structure from ouabain, because ouabain shows an UV absorbance maximum at 220 nm while the new inhibitor at 250 nm.展开更多
Toad venom, called as Chan Su in China, is a widely used traditional Chinese medicine(TCM) whose active components are mainly bufadienolides. Chan Su could exhibit cardiotonic, anti-microbial, anti-inflammatory and, m...Toad venom, called as Chan Su in China, is a widely used traditional Chinese medicine(TCM) whose active components are mainly bufadienolides. Chan Su could exhibit cardiotonic, anti-microbial, anti-inflammatory and, most importantly, anti-cancer effects. In the present review, reports about the in vitro, in vivo and clinical anti-cancer effects of Chan Su or its representative component, bufalin, were summarized.And, reported anti-cancer mechanisms of cardenolides, structure analogues of bufadienolides, were also introduced. Based on the results got from research of Chan Su/bufalin and the results from cardenolides, possible signal network related to the anti-cancer effects of Chan Su/bufalin was predicted. Furthermore, future potential use of Chan Su in anti-cancer therapy was discussed.展开更多
目的观察华蟾素对人乳腺癌MCF-7、MDA-MB-231细胞人第10号染色体缺失的磷酸酶和张力蛋白的同源基因(phosphatase and tensin homolog deleted on chromosome ten,PTEN)蛋白表达的影响。方法将MCF-7、MDA-MB-231细胞按随机数字表法分为...目的观察华蟾素对人乳腺癌MCF-7、MDA-MB-231细胞人第10号染色体缺失的磷酸酶和张力蛋白的同源基因(phosphatase and tensin homolog deleted on chromosome ten,PTEN)蛋白表达的影响。方法将MCF-7、MDA-MB-231细胞按随机数字表法分为对照组、高剂量组、低剂量组。低、高剂量组分别加入0.5、5.0 mg/ml华蟾素注射液l00μl/孔进行干预;对照组加入等体积的RPMI-1640培养液。干预后6、12、24、48、72 h,采用细胞计数试剂盒-8法检测细胞增殖率,干预后72 h,采用Western blot法检测蛋白激酶B/哺乳动物雷帕霉素靶蛋白(protein kinase B/mammalian target of rapamycin,AKT/mTOR)通路蛋白及p-PTEN蛋白表达。结果与对照组比较,干预后24、48、72 h,高剂量组MCF-7、MDA-MB-231细胞增殖率降低(P<0.05);低、高剂量组MCF-7、MDA-MB-231细胞p-AKT[MCF-7:(0.357±0.064)、(0.215±0.056)比(0.924±0.085);MDA-MB-231:(0.310±0.022)、(0.194±0.019)比(0.811±0.089)]、p-mTOR[MCF-7:(0.476±0.039)、(0.217±0.038)比(0.838±0.058);MDA-MB-231:(0.300±0.031)、(0.223±0.025)比(0.896±0.096)]、p-S6[MCF-7:(0.551±0.068)、(0.428±0.041)比(1.254±0.264);MDA-MB-231:(0.281±0.014)、(0.197±0.012)比(0.748±0.022)]、p-PTEN[MCF-7:(0.487±0.170)、(0.184±0.135)比(1.003±0.284);MDA-MB-231:(0.261±0.184)、(0.170±0.105)比(1.014±0.206)]表达降低(P<0.05)。结论华蟾素可抑制MCF-7、MDA-MB-231细胞增殖,推测可能与其下调AKT/mTOR信号通路上游蛋白p-PTEN蛋白表达有关。展开更多
基金Supported by the National Natural Science Foundation of China(Nos.30772721 and 30873360)
文摘In vitro models of human colon carcinoma cell line(Caco-2 cell monolayer) and human intestinal bacteria were used to investigate the intestinal transport and biotransformation of resibufogenin and cinobufagin in Chan Su by HPLC/APCI-MSn. The experimental results of Caco-2 cell monolayer demonstrate that the apparent permeability coefficients(Papp) of resibufogenin and cinobufagin are higher than 10–6 cm/s, which indicates that both resibufogenin and cinobufagin have a good absorption in the small intestine. And the biotransformation result of human intestinal bacteria shows that resibufogenin has been transformed to 3-epiresibufogenin and cinobufagin has been transformed to 3-epicinobufagin, deacetylcinobufagin and 3-epideacetycinobufagin, respectively.
文摘An endogenous inhibitor of the sodium pump from the toad venom Chan′su was purified. The dry substance Chan′su was extracted with methanol. The dry residue was passed subsequently through membrane filters with the exclusion size of 10 000, 3 000 and 1 000 at 1 psi in a Filtron Pro Vario-3-System and applied to thin layer chromatography made of Silica 60 F 254+366 separated in CHCl 3∶MeOH∶H 2O. The R f 0.55 fractions inhibiting the sodium pump were purified on a HPLC-RP C18 using a linear H\-2O/methanol gradient at 220 nm and 250 nm on DAD detector. The bio-activity was measured by a specific enzyme-linked immunosorbent assay (ELISA) and 86 Rb-uptake into human red blood cells. In Chan′su there exists an inhibitor of the sodium pump which interacts with antibodies against ouabain and proscillaridin A, and inhibits the sodium pump activity in the red blood cells. The results show that a low molecular weight compound has been isolated from the toad venom Chan′su. It inhibits the sodium pump, interacts preferentially with antibodies against ouabain. It is water soluble and shows a maximum of absorbance at 250 nm. Several milligrams of the compounds from one kilogram were obtained in pure form. The inhibitor differs considerably in its chemical structure from ouabain, because ouabain shows an UV absorbance maximum at 220 nm while the new inhibitor at 250 nm.
基金Partly supported by the National Science&technology Major Project of China(2014ZX09301-306-03)Shanghai Science&Technology Support Program(13431900401)the National Nature Science Foundation of China(81373964)
文摘Toad venom, called as Chan Su in China, is a widely used traditional Chinese medicine(TCM) whose active components are mainly bufadienolides. Chan Su could exhibit cardiotonic, anti-microbial, anti-inflammatory and, most importantly, anti-cancer effects. In the present review, reports about the in vitro, in vivo and clinical anti-cancer effects of Chan Su or its representative component, bufalin, were summarized.And, reported anti-cancer mechanisms of cardenolides, structure analogues of bufadienolides, were also introduced. Based on the results got from research of Chan Su/bufalin and the results from cardenolides, possible signal network related to the anti-cancer effects of Chan Su/bufalin was predicted. Furthermore, future potential use of Chan Su in anti-cancer therapy was discussed.
