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Abstract The locust bean gum (LBG) is a polysaccharide with thickening, stabilizing and gelling properties and it has been used in the preparation of pharmaceutical formulations. Hydrogels (HGs) are obtained from natural or synthetic materials that present interesting properties for skin application. This study aimed to develop HGs from LBG using indole-3-carbinol (I3C) as an asset model for cutaneous application. HGs were prepared by dispersing LBG (2%, 3% and 4% w/v) directly in cold water. The formulations showed content close to 0.5 mg/g (HPLC) and pH ranging from 7.25 to 7.41 (potentiometry). The spreadability factor (parallel plate method) was inversely proportional to LBG concentration. The rheological evaluation (rotational viscometer) demonstrated a non-Newtonian pseudoplastic flow behavior (Ostwald De Weale model), which is interesting for cutaneous application. The HET-CAM evaluation showed the non-irritating characteristic of the formulations. The bioadhesive potential demonstrated bioadhesion in a concentration-dependent manner. Permeation in human skin using Franz cells showed that the highest LBG concentration improved the skin distribution profile with greater I3C amounts in the viable skin layers. The present study demonstrated the feasibility of preparing HGs with LBG and the formulation with the highest polymer concentration was the most promising to transport active ingredients through the skin.
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Polissacarídeos/análise , Borracha/análise , Hidrogéis/análise , Potenciometria/instrumentação , Preparações Farmacêuticas/administração & dosagem , Cromatografia Líquida de Alta Pressão/métodos , Creme para a Pele/classificaçãoRESUMO
Indole-3-carbinol(I3C),an important anticancer compound found in broccoli,has attracted considerable attention.The rapid extraction and accurate analysis of I3C in the pharmaceutical industry in broccoli is challenging as I3C is unstable at low pH and high temperature.In this study,a rapid,accurate,and low-cost ultrasound-assisted dispersive-filter extraction(UADFE)technique based on poly(deep eutectic solvent)-graphene oxide(PDES-GO)adsorbent was developed for the isolation and analysis of I3C in broccoli for the first time.PDES-GO with multiple adsorption interactions and a fast mass transfer rate was synthesized to accelerate adsorption and desorption.UADFE was developed by combining dispersive solid-phase extraction(DSPE)and filter solid-phase extraction(FSPE)to realize rapid extraction and separation.Based on the above two strategies,the proposed PDES-GO-UADFE method coupled with high-performance liquid chromatography(HPLC)allowed the rapid(15-16 min),accurate(84.3％-96.4％),and low-cost(adsorbent:3.00 mg)analysis of I3C in broccoli and was superior to solid-phase extraction,DSPE,and FSPE methods.The proposed method showed remarkable linearity(r=0.9998;range:0.0840-48.0 μg/g),low limit of quantification(0.0840 μg/g),and high precision(relative standard deviation≤5.6％).Therefore,the PDES-GO-UADFE-HPLC method shows significant potential in the field of pharmaceutical analysis for the separation and analysis of anti-cancer compounds in complex plant samples.
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Two skeletally undescribed polyketide-indole hybrids (PIHs), named indolchromins A and B, were generated from indole-3-carbinol (I3C) in the fungal culture (). The indolchromin structures were elucidated mainly by their 1D and 2D NMR spectra with the former confirmed by the single-crystal X-ray crystallographic analysis. Each indolchromin alkaloid was chirally separated into four isomers, whose absolute configurations were assigned by comparing the recorded circular dichroism (CD) spectra with the electronic CD (ECD) curves computed for all optional stereoisomers. Furthermore, the indolchromin construction pathways in fungal culture were clarified through enzyme inhibition, precursor feeding experiment, and energy calculation. The cascade reactions, including decarboxylative Claisen condensation catalyzed by 8-amino-7-oxononanoate synthase (AONS), C()-H activation, double bond migration, and Michael addition, all undergone compatibly during the fungal cultivation. In an MIC range of 1.3-8.6 μmol/L, (2,4)- and (2,4)-indolchromin A and (2,4)-indolchromin B are inhibitory against , , sp., , and . (2,4)-Indolchromin A and (2,4)-indolchromin B were cytotoxic against the human breast cancer cell line MDA-MB-231 with IC values of 27.9 and 131.2 nmol/L, respectively, with the former additionally active against another human breast cancer cell line MCF-7 (IC 94.4 nmol/L).
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Objective To investigate the protective effect of indole-3-carbinol (I3C) on radiation-induced mouse bone marrow hematopoietic cell injury and the involved mechanisms. Methods (1) The bone marrow nuclear cells (BMNCs) from CD45.1 subtype of C57BL/6J mice were collected by a density gradient centrifugation method. The BMNCs were pretreated with a series doses of I3C (0 mol/L, 10-8 mol/L-10-3 mol/L) and then exposed with radiation of 137Csγ-ray (doses of irradiation were 0 Gy, 1 Gy and 4 Gy). After 18-hour culturing, the bioluminescence method was used to detect the cell viability. (2) These cells were divided into control group and 10-6 mol/L I3C group. Both groups were received the irradiation (0 Gy, 1 Gy and 4 Gy) and inoculated into the methylcellulose semi-solid culture medium to incubate 7 days, the colony forming unit-granulocyte monocytes (CFU-GM) were observed. (3) Twenty-four CD45.2 subtype mice used as the receptor were exposed with 8 Gy radiation. The CD45.1 BMNCs were divided into control group, 4 Gy irradiation group, 4 Gy irradiation and 10-6 mol/L I3C group. Donor cells were harvested from C57BL/6J (CD45.1) mice after they received various treatments, and were then mixed with competitive BMNCs from C57BL/6J (CD45.2) mice. The mixed cells were transplanted into recipient mice (8 mice/group). Flow cytometry was used to analyze the proportion of donor cells in peripheral blood of receptor. (4) The cells were divided into control group, 10-6 mol/L I3C group, 1 Gy irradiation group, 1 Gy irradiation with 10-6 mol/L I3C group. After 24-hour culturing, Western blot assay was used to detect the expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1). Results (1) I3C showed a significant cytotoxic effect on the BMNCs when its concentration was above 10-4 mol/L. 10-7-10-6 mol/L I3C could reduce the radiation injury of BMNCs under the same dose of irradiation. Therefore, 10-6 mol/L I3C was chosen for subsequent experiments. (2) The CFU-GM was significantly higher in 10-6 mol/L I3C group than that of control group (P<0.05). (3) Results of flow cytometry showed that the proportion of donor cells in receptor was significantly higher in 4 Gy irradiation group than that of control group, which decreased the engraftment capability of irradiated HSCs (P<0.05), although the engraftment capability of irradiated HSCs improved after 10-6 mol/L I3C treatment. (4) I3C significantly enhanced the increased protein expression of Nrf2 and HO-1 caused by radiation (P<0.05). Conclusion I3C has a protective effect on hematopoietic cells following radiation-induced injury, which may be related with activating the Nrf2/HO-1 signal pathway.
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Objective To study the radioprotective effect of indole-3-carbinol (I3C) acid condensation products.Methods Cell colony formation assay was used to determine cell survival rate,and Western Blot assay was employed to measure protein expression.Results Seven kinds of the I3C acid condensation products showed different radioprotective effect on normal fibrous epithelial cells 184A1,among which 24 h pre-treatment of CTET (1 μmol/L),LTET (1 μmol/L),HI-IM (1 μmol/L) and 3,3'-diindoly methane (DIM) (0.3 μmol/L) showed significant increase of cell survival rate following irradiation with γ-ray,and the difficence was statistically significant (P＜0.05,P＜0.01).However,CT (1 μmol/L),LTr-1 (1 μmol/L) and ICZ (1 μmol/L) showed no effect on cell survival rate caused by radiation (P＞0.05).Furthermore,CTET,LTET,HI-IM and DIM activated the phosphorylation of ATM,BRCA1 and NBS1 proteins.HI-IM significantly decreased radiation-caused cell death and apoptosis.Conclusions CTET,LTET,HI-IM,and DIM can significantly reduce the radiosensitivity in 184A1 cells,and the mechanism may be related to the regulation of DNA damage and the repair of protein phosphorylation.
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Epidermal Growth Factor Receptor (EGFR) is one of the four members of the Human Epidermal Receptor (HER) family, which is deregulated and over expressed in platinum resistant ovarian cancer. Thus, targeting EGFR receptor along with platinum drugs is one of the major strategies to increase the platinum drug sensitivity. That‟s why, in this study, we aimed to investigate the inhibitory activity and binding site analysis of indole-3-carbinol and its active metabolite 3,3'-diindolylmethane by using molecular simulation studies, also metabolic profile had been investigated by SOM prediction. The 3,3'-diindolylmethane showed significant inhibitory activity and binding energy comparing to indole-3-carbinol, also it processed lower toxicity and will undergo aromatic hydroxylation due its high intrinsic activity and Fe accessibility. Though our research study supports previous reports of EGFR inhibition, further in vivo study is necessary for validation of toxicological and pharmacokinetic study. However, the current work tries to address most of the variables in the dynamic drug design process by In silico study in order to boost the potentiality of the selected molecule to serve as good leads in terms of optimum pharmacokinetic and toxicological attributes.
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OBJECTIVE:To study the inhibitory effects of phosphatidylinositol 3-kinase (PI3K) inhibitor LY29400 (“LY”) combined with indole-3-carbinol (I3C) on the proliferation of human anaplastic thyroid carcinoma FRO cells. METHODS:RFO cells in logarithmic growth phase were divided into blank control group,I3C (250 μmol/L) group,LY (10 μmol/L) group and combination (I3C of 250 μmol/L + LY of 10 μmol/L) group,which were subject to 24,48 and 72 h drug action. MTT method was used to determine and calculate the inhibitory rates of the cells in all groups,flow cytometry assay to determine the apoptosis rates after 48 h action,Western blot method to determine the expressions of Capspase-3 proteins thereafter and immunohistochemi-cal method to determine the expressions of Bcl-2 and Bax thereafter. RESULTS:The inhibitory rate obviously increased with the ex-tension of drug action time,with statistical significance at each time point in combination group (P<0.05). Compared to I3C group and LY group,combination group had higher inhibitory rate,apoptosis rate and the expressions of Capspase-3 protein and Bax,and lower Bcl-2 expression and ratio of Bcl-2 to Bax,with statistical significance (P<0.01). CONCLUSIONS:LY com-bined with I3C has synergistic inhibitory effects on the proliferation of FRO cells by a mechanism which may be related to apopto-sis induced by down-regulating Bcl-2 expression,up-regulating Bax expression and activating Caspase cascade reaction.
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Induction of apoptosis in target cells is a key mechanism by which chemotherapy promotes cell killing. The purpose of this study was to determine whether Indole-3-Carbinol (I3C) and Genistein in combination with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induce apoptosis in endometrial cancer cell (Ishikawa) and to assess apoptotic mechanism. The MTT assay and flow cytometry were performed to determine cell viability and cell cycle. The induction of apoptosis was measured by caspase-3 activity test, DNA fragmentation assay, annexin V binding assay and western blot analysis. There was no effect in cell growth inhibition and cell cycle progression alone or in two-combination. However, the treatment of I3C and Genistein followed by TRAIL showed significant cell death and marked increase in sub-G1 arrest. Three-combination treatment revealed elevated expression of DR4, DR5 and cleaved forms of caspase-3, caspase-8, PARP. The Flip was found down regulated. Moreover, increase in caspase-3 activity and DNA fragmentation indicated the induction of apoptosis. The results indicate that I3C and Genistein with TRAIL synergistically induced apoptosis via death receptor dependent pathway. Our findings might provide a new insight into the development of novel combination therapies against endometrial cancer.
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Feminino , Humanos , Anticarcinógenos/farmacologia , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Caspase 8/metabolismo , Linhagem Celular Tumoral , Sinergismo Farmacológico , Neoplasias do Endométrio/metabolismo , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos dos fármacos , Genisteína/farmacologia , Indóis/farmacologia , Poli(ADP-Ribose) Polimerases/metabolismo , Receptores do Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Ligante Indutor de Apoptose Relacionado a TNF/farmacologiaRESUMO
Background Ultraviolet radiation is one of factors of the formation of age-related cataract.Indole-3-carbinol(I3C) is a plant chemical material with inhibitory effect on oxidative-induced cell damage and formation of amyloid fibrils,and the oxidative damage and amyloid fibrils are associated with cataract.However,the relationship between I3C and α-crystallin is in study. Objective This study was to evaluate the effects of ultraviolet-B laser irradiation on the secondary structure of α-crystallin and to explore the protection of I3C to chaperone activity of α-crystallin. Methods The fresh eyeballs were obtained from 1-year-old cattle to prepare the purified lens α-crystallin by gel chromatography.α-Crystallin was isolated from cattle lenses using gel chromatography.The purified α-crystallin was collected using fast protein liquid chromatography ( FPLC ) and exposed to 1:308 nmultraviolet-B at different irradiation intensities ( 23.75,118.75,475.00,1187.50,2375.00,4750.00,11 875.00,23 750.00 mJ/cm2 ) and then to ultraviolet-B 2:308 nm with irradiation intensities of 28 535.00,6730.00,3435.00,1910.00,1040.00 mJ/cm2.Ultraviolet-absorbance spectra,tryptophan fluorescence and N-formylkynurenine (N-FK)fluorescence spectra of both irradiated and non-irradiated α-crystallin were measured.I3C at the concentrations of 50 μmol/L and 100 μmoL/L were added to the α-crystallin solution to perform a catalase (CAT) thermal aggregation to confirm the chaperone activity of the α-crystallin,and the α-crystallin solution without any I3C was used as control.The ratios of A360 between various intervene groups with control group were calculated using spectrophotometry.Results The A280 values of the α-crystallin declined to 10％ at the ultraviolet-B irradiation intensity of 1187.5 mJ/cm2 and that at the intensity of 23.75 J/cm2 lowed to 2％.A negative correlation was seen between the ultraviolet-B irradiation intensity and the A280 value of the α-crystallin (R2=0.925 ) and a positive correlation was found between ultraviolet-B with N-FK ( R2 =0.949 ).Ultraviolet-B irradiation intensity showed a negative correlation with Trp fluorescence intensity (R2 =0.996 ).CAT hot condensed experiment revealed that after addition of different concentrations of indole-3-carbinol,the relative A360 values at various ultraviolet-B irradiation group were significantly higher than those of the control group (P =0.000),and the decreasing degree of chaperone activity of α-crystallin was lower than that of the control group ( P =0.000 ). Conclusions The study suggests that I3C can protect the chaperone activity of α-crystallin from photooxidation,and the ultraviolet-B laser may be a good exposure source compared with ultraviolet lamp.The ultraviolet-B laser irradiation causes the alteration of structure and chaperone activity of α-crystallin.
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Indole-3-carbinol (I3C) found in various cruciferous vegetables has been shown to exert anti-carcinogenic activity in several target organs. Our study was conducted to assess the modifying effect of I3C on the development of colon tumor induced by azoxymethane (AOM). Eighty-seven male F344 rats were divided into 5 groups and were treated with AOM followed by I3C 100 or 300 ppm, AOM alone, I3C alone, and non-treatment, respectively. The animals were subcutaneously injected with AOM. Then diet containing I3C were fed to the rats for 37 weeks. All rats were sacrificed at 40 weeks. Liver and kidney weights of rats treated with I3C at doses of 100 or 300 ppm were significantly increased compared to those of the control group. Colonic tumor incidence and multiplicity of rats treated with I3C at doses of 100 and 300 ppm were not significant compared to those of AOM alone group. In the pathological examination, most of tumors were classified with adenoma and adenocarcinoma in the small and large intestine. These results demonstrated that I3C may have not chemopreventive effect on the rat colon carcinogenesis.
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Animais , Humanos , Masculino , Ratos , Adenocarcinoma , Adenoma , Azoximetano , Colo , Dieta , Incidência , Indóis , Intestino Grosso , Rim , Fígado , Ratos Endogâmicos F344 , Verduras , Pesos e MedidasRESUMO
OBJECTIVE: To determine whether Indole-3-carbinol (I3C) can enhance the inhibitory effect of genistein on a human uterine leiomyoma cells. METHODS: Five uterine leiomyoma tissues were obtained from hysterectomies conducted on the benign diseases and cultured primarily. MTS reduction assay was carried out to determine the viability of human uterine leiomyoma cells. Cell cycle analysis for I3C and genistein treated human uterine leiomyoma cells was done by Fluorescent activated cell sorter (FACS) analysis. To detect the presence and expression of cell cycle related proteins was done by Western blot analysis. RESULTS: I3C and genistein induced growth inhibition in a dose dependent manner, treatment with 100 micro mol/L I3C and 100 micro mol/L genisten blocked 60% cell growth. FACS results showed that treatment with the I3C and genistein increased the percentage of cells in G2/M phase and decreased S phase. From Western blot analysis it revealed I3C and genistein induced the expression of p53, p21, and p27 increasing. Reduced expression of cyclin B1 and cyclin E were detected in treatment with I3C and genistein. The expression levels of these proteins correlate with G2/M cell cycle arrest. Activation of caspase pathway and fragmentation of PARP did not take place. CONCLUSIONS: These results demonstrate that I3C enhances genistein-mediated uterine leiomyoma cell growth inhibition through the cell cycle arrest at G2/M phase by decreasing the production of cyclin B1. Because of the synergistic effect of I3C and genistein, the potential exists for the therapeutic efficacy of each phytochemical when used in combination.
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Humanos , Western Blotting , Ciclo Celular , Pontos de Checagem do Ciclo Celular , Ciclina B1 , Ciclina E , Ciclinas , Genisteína , Histerectomia , Leiomioma , Fase SRESUMO
Objective To observe the effects of indole-3-carbinol on neointimal hyperplasia and restenosis of rat artery after balloon injury and the possible mechanisms.Methods Balloon dilation was used to establish the neointimal injury model of left carotid artery in rats.Twenty Sprague-Dawley rats were randomly divided into single balloon dilation group(control group)and balloon dilation followed by indole-3-carbinol therapy group(therapeutic group).After balloon dilation,indole-3-carbinol(12.5,25,50 mg/d)was applied to the rats for 7 days respectively.The rats were killed two weeks after balloon dilation and the injured vascular specimens were harvested for pathologic examination and immunohistochemical staining.Results ①The neointimal thickness,neointimal area in the therapeutic groups were significantly less than that of the control group(P
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AIM To study the hepato-protection of indole-3-carbinol (I3C), one of glucosinolate compounds in cruciferous vegetables, on ethanol-induced hepatotoxicity. METHODS Ethanol 50 mmol?L -1 was used to induce hepatotoxicity in precision liver-cut slices of rats. The release of glutamic pyruvic transaminase(GTP), glutamic oxaloacetic transaminase (GOT), lactate dehydrogenase (LDH) and glutathione S-transferase (GST) into culture medium, and aniline hydroxylase (ANH) and alcohol dehydrogenase (ADH) in cytosol of hepatocytes in the precision liver-cut were measured with different doses of I3C. The histopathological observation of liver slices was also done to evaluate the effect of I3C. RESULTS The release of GTP, GOT, GST and LDH in the medium increased significantly when the slices were treated with 50 mmol?L -1 ethanol for 4 h. I3C at concentrations of 100～400 ?mol?L -1 effectively reduced the release of GTP, GOT, GST and LDH in the medium of ethanol-treated slices. Meanwhile, the activities of cytosol ANH and ADH decreased to the normal levels. The histopathological changes of liver slices induced by ethanol were lessened by I3C treatment. CONCLUSION I3C can markedly protect rat liver slices from ethanol-induced hepatotoxicity, which partly results from the change of ethanol metabolism in liver.
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Aim To investigate effects of indole-3-carbinol (I3C) on hepatic stellate cells (HSCs) activated by acetaldehyde in precision-cut liver slices (PCLS). Methods PCLS were incubated with 700 ?mol?L-1 acetaldehyde and 200 ~ 800 ?mol?L-1 I3C for 6 h. The expression of ?-smooth muscle actin(?-SMA) in liver slices was analyzed by immunohistochemistry. The leakages of glutathione S-transferase (GST), lactate dehydrogenase (LDH) and content of transforming growth factor-?1 (TGF-?1) in media were assayed. Contents of malondialdehyde (MDA) and hydroxyproline (Hyp) in tissue were also determined. Expressions of matrix metalloproteinases-1 (MMP-1) and tissue inhibitor of metalloproteinase (TIMP-1) in media were analyzed by the western blot. Results The increase of activated HSCs due to acetaldehyde was inhibited by I3C (200~800 ?mol?L-1). Meanwhile, I3C treatment (200~800 ?mol?L-1) showed significant and concentration-dependent antagonistic actions on the increment of GST, LDH leakages into the media and MDA, Hyp contents in tissues induced by acetaldehyde. The increase of TGF-?1 was also remarkable inversed by I3C (200~800 ?mol?L-1). As compared with acetaldehyde group, the ratio of MMP-1/TIMP-1 was increased significantly by I3C treatment (800 ?mol?L-1)(P