Effect of solanine on radiosensitivity of osteosarcoma cells / 中华放射医学与防护杂志

Objective To investigate the effect of solanine on radiosensitivity of osteosarcoma cells. Methods Osteosarcoma cells were treated with solanine, at the same time, radiation treatment was given, cell proliferation was measured by MTT assay, cell apoptosis was measured by flow cytometry, the protein levels of Ki-67, PCNA, Bax and Cleaved Caspase-3 were detected by Western blot and mitochondrial membrane potential was detected by JC-1 method, the ROS level in cells was detected by DCFH-DA method. Radiosensitivity was measured by plate cloning. Results Compared with cells that were not treated with solanine and radiation, the proliferation of osteosarcoma cells and the Ki-67 and PCNA protein levels were decreased after solanine or radiotherapy ( F=55. 165, 50. 667, 23. 389, P<0. 05) ,the apoptotic rate and the level of Bax and Cleaved Caspase-3 protein increased ( F=54. 588, 42. 924, 51. 541, P<0. 05), the mitochondrial membrane potential decreased (F=40. 762, P<0. 05), and the ROS level in the cells increased( F=79. 055, P<0. 05) . Compared with cells treated with solanine or radiotherapy, the proliferation of osteosarcoma cells and the levels of Ki-67 and PCNA proteins were decreased after solanine combined with radiation treatment ( F=55. 165, 50. 667, 23. 389, P<0. 05) , the apoptotic rate and the levels of Bax and Cleaved Caspase-3 were increased ( F=54. 588, 42. 924, 51. 541, P<0. 05), the mitochondrial membrane potential was decreased (F=40. 762, P<0. 05), and the ROS level was increased (F=79. 055, P<0. 05). The radiosensitization ratio of solanine to osteosarcoma cells was 1. 786. Conclusions Solanine inhibits proliferation and induces apoptosis of osteosarcoma cells, and improves radiosensitivity of osteosarcoma cells.

Ecofriendly Synthesis of Silver Nanoparticles Using Potato Steroidal Alkaloids and Their Activity Against Phytopathogenic Fungi

ABSTRACT In order to reduce the excessive reliance on the toxic chemical fungicides, the present study aimed to isolate the total potato glycoalkaloids (TPAs), and the two steroidal alkaloids α-chaconine and α-solanine from potatoes, Solanum tuberosum L. Their structures were characterized using physical and spectroscopic methods including (UV, IR, 1H, 13C--NMR, 2D 1H-1H COSY, HMBC and NOESY). Silver nanoparticles (AgNPs) were prepared from potato alkaloids through a green synthesis approach. Potato alkaloids and their nanoparticles inhibited mycelial growth of the phytopathogenic fungi Alternaria alternate, Rhizoctonia solani, Botrytis cinerea and Fusarium oxysporum f. sp. lycopersici with low minimal inhibitory and minimal fungicidal concentrations. R. solani was the most susceptible, while F. oxysporum was the most resistant. TPAs was the most fungitoxic (EC50's were 19.8, 22.5, 26.5 and 32.3 µg/ml against R. solani, A. alternate, B. cinerea and F. oxysporum respectively). A mixture of α-solanine and α-chaconine (1:1) showed a marked antifungal activity. AgNPs (size 39.5-80.3 diameter) from alkaloids showed improved fungitoxic activity (EC50's of TPAs nanoparticles ranged between 10.9 and 16.1 µg/ml). Alkaloids exhibited no or a slight phytotoxicity against wheat and radish. Results recommend the potential of using potato alkaloids and their nanoparticles as biorational alternatives to conventional fungicides.

Identification of Unknown Toxicants in Food Poisoning by High Resolution Mass Spectrometry-Data Independent Acquisition-Post Targeted Screening Strategy / 分析化学

Based on ultra-high-performance liquid chromatography-quadrupole/time of flight mass spectrometry (UPLC-Q-TOF MS/MS),a post targeted screening strategy using high resolution mass spectrometry under data independent acquisition was established,and successfully applied to screen and identify unknown toxicants in food poisoning patients' vomit and feces.After solvent extraction,the samples were separated and analyzed on a reversed-phase C18column using a gradient elution program of 5 mmol/L ammonium formate and 0.1% formic acid aqueous solution(A) and 0.1% formic acid in acetonitrile solution (B).All-ions MS/MS information of samples was obtained using Q-TOF MS(ESI+) in MSE mode,followed by peak recognition and library searching using the UNIFI Scientific Information System,as a result,suspected toxic compound α-solanine was primarily detected.Furthermore,the related chemicals including hydrolysates and metabolites were identified in both samples based on post-targeted screening strategy,such as α-chaconine,β-chaconine,γ-solanine/chaconine and solanidine.The concentration of α-solanine in both samples was approximately 0.1 mg/kg by one-point calibration method,the total amount of solanine analogues in two samples was about 0.5 mg/kg and 0.6 mg/kg respectively through normalization of peak area.The high resolution mass spectrometry screening strategy established here could provide efficient screening method for rapid detection and identification of toxicants in unknown food poisoning.

Effect and mechanism of Solanine on proliferation and apoptosis of Jurkat cells / 中华实用儿科临床杂志

Objective To explore the effect of proliferation and apoptosis of Solanine on acute T lymphocyte leukemia (T-ALL) Jurkat cells and its mechanism.Methods After treated with different concentrations of Solanine,the proliferation of Jurkat cells was detected by CCK-8 assay,and the effect of Solanine on apoptosis of Jurkat cells were detected by flow cytometry.The expression of Bcl-2 and Bax in Jurkat cells were detected by Western blot,and the expression levels of Bcl-2 mRNA and Bax mRNA were detected by real-time fluorescence quantitative polymerase chain reaction.Results CCK-8 assay showed that Solanine significantly inhibited the proliferation of Jurkat cells in a dose-and time-dependent manner.The results of flow cytometry showed that the apoptosis rates of Jurkat cells treated with Solanine for 24 h were (2.40-± 0.98) ％,(28.43-± 4.86) ％,(41.56-± 1.87) ％,respectively,in a dose-dependent manner.Western blot showed that Solanine could increase the expression of Bax and decrease the expression of Bcl-2 in Jurkat cells,and they all were dose-dependent.Conclusion Solanine can significantly inhibit the proliferation and induce apoptosis of Jurkat cells.The mechanism is related to the up-regulation of Bax expression and down-regulation of Bcl-2 expression.

Effect of solanine on proliferation and apoptosis of U251 cells / 中草药

Objective: To investigate the effect of solanine on growth and proliferation of U251 cells. Methods: U251 cells were cultured with different concentration of solanine together. CCK-8 method, wound healing assay, Transwell method, Hochest assay, flow cytometry screening and Western blotting were used to detect inhibitory rate, migration and invasive rates, apoptosis rate and expression level of cell apoptosis-related proteins. Results: CCK-8 assay showed the median inhibition concentration (IC50) of 48 h was 20.05 μg/μL. It was effective that solanine in the concentration range of 5-35 μg/μL could inhibit the growth and proliferation of U251 cells, and played a role in a dose-dependent manner (P < 0.05). Compared with control group, experimental groups can not only inhibit U251 cell migration and invasion in a dose-dependent manner (P < 0.05), but also appear some significant apoptosis characteristics with a concentration dependence. Western blotting assayed that the expression of Bax protein was upregulated, whereas Bcl-2 protein expression was downregulated in experimental groups compared with control group and these changes were dose-dependent. Conclusion: Solanine can inhibit growth and proliferation of U251 cells in effective concentration and in a dose-dependent manner. Solanine can inhibit the invasion and migration of U251 cells. Solanine can induce the apoptosis of U251 cells, and inhibit the proliferation and growth of U251 cells via regulating the expression of Bax and Bcl-2 proteins and affecting Bcl-2/Bax ratio.

Reversal effect of solanine on multidrug-resistance of leukemia K562/ADR cells and its mechanism / 肿瘤

Objective: To investigate the effect of solanine on multidrug resistance of leukemia K562/ADR cells, and to explore its mechanism. Methods: K562/ADR cells were treated with different concentrations (5-40 μg/mL) of solanine alone or in combination with doxorubicin (DOX) for 24 h. The intracellular toxicity and DOX-sensitization effect of solanine were detected by CCK-8 assay. The cell-associated mean fluorescence intensity of DOX was detected by FCM method. The expressions of multidrug resistance-associated protein 1 (MRP1), c-Jun NH2-terminal kinase (JNK), and phosphorylated JNK (p-JNK) were determined by Western blotting. Results: Solanine at non-toxic doses (5 and 10 μg/mL) significantly enhanced the cytotoxicity of DOX (both P < 0.05), and significantly increased the mean fluorescence intensity of DOX in K562/ADR cells in a dose-dependent manner (both P < 0.05). After treatment with 5 and 10 μg/mL solanine, the expression levels of MRP1 and p-JNK proteins were significantly decreased (all P < 0.05) in K562/ADR cells. Conclusion: Solanine can reverse multidrug resistance of K562/ADR cells in vitro. The mechanism may be related to blocking JNK signaling pathway and downregulating MRP1 expression.

Solanine induces the apoptosis of human prostate cancer cells via ROS/p38 signaling pathway / 西安交通大学学报(医学版)

Objective To investigate the molecular mechanism of solanine-induced apoptosis of prostate cancer cells Du145 and LNCaP.Methods The effects of solanine on the viability of Du145 and LNCaP cells were evaluated by MTT assay.The generation of intracellular reactive oxygen species (ROS) and solanine-induced apoptosis were measured by flow cytometry.The protein levels of p38 and p-p38 expressions were examined by Western blot.Results Solanine significantly inhibited the viability of Du145 and LNCaP cells in a dose-dependent manner (P＜0.01).The inhibition of solanine on cell viability was suppressed by the ROS scavenger NAC.ROS generation,apoptosis and phosphorylation of p38 were induced by treatment with solanine at 40 μmol/L for 24 h.The expression of p38 and solanine-induced apoptosis were suppressed by NAC and SB203580.Conclusion Solanine induces the apoptosis of human prostate cancer cell via the RO.S-p38 signaling pathway.

Técnica de Tinta China en células adherentes en cultivo / China ink in adherents ceil in culture

Objetivo. Implementar la utilización de Tinta China como alternativa para visualizar, cambios a nivel de matriz y pared celular, en células vivas adheridas en cultivo, antes y después de la exposición a una sustancia toxica. Métodos. Se implementó la Tinta China, como técnica de contraste en microscopia óptica, comparando la nitidez observada (adecuada apreciación del borde de las estructuras) entre frascos de cultivo de células troncales de médula ósea de rata (CTMO) expuestos al glicoalcaloide tóxico α-solanina. Las diferencias de nitidez se compararon entre los diversos tratamientos, con test exacto de Fisher. Resultados. La tinción con Tinta China permitió identificar con nitidez los cambios fenotípicos celulares anormales, secundarios a la exposición del citotóxico en células adherentes en cultivo (p <0.0001). Conclusiones. La Tinta China es útil en la visualización nítida de las células CTMO y de los efectos producidos por el glicoalcaloide α-solanina en células adheridas en cultivo. Es un método sencillo que aporta al entendimiento del efecto que diversas sustancias producen en las CTMO en cultivo.

Objective. To implement the use of China ink, as an alternative technique, to display morphological changes in cellular wall and cellular matrix in adherent living cells in culture flask, before and after exposure to a toxic substance. Methods. China Ink was implemented as a contrast technique in optical microscopy, by comparing observed sharpness (proper appreciation of edge structures) from cultures of bone marrow stem cells (CTMO) exposed to the toxic glycoalkaloid α-solanine. The sharpness differences were compared among the diverse treatments with Fisher's exact test. Results. China Ink staining clearly helps to identify abnormal phenotypic changes, secondary to cytotoxic exposure in adherent cells in culture (p <0.0001). Conclusions. China Ink is useful in clearly displaying the CTMO cells and the effects of the α-solanine glycoalkaloid in adherent cells in culture. It is a simple method that contributes to the understanding of the effect of various substances on CTMO in culture.

Effect of angiogenesis on Solanine and VEGF antibody in chicken embryo transplantation model of human colon cancer cells / 重庆医学

Objective To establish model of the chicken embryo transplantation of human colon cancer cells ,and investigate the effect of Solanine、VEGF antibody and Solanine combined with VEGF antibody on human colon cancer cells induce tumor angio‐genesis and tumor proliferation .Methods The model of the chicken embryo transplantation of human colon cancer HT‐29 cells were divided into three experimental group and control group .We added to the chick embryo chorioallantoic membrane with Sola‐nine、VEGF antibody and Solanine+ VEGF antibody mixture ,PBS was added to the control group .Then we analysed picture through the stereomicroscope and IPP 6 .0 image analysis software ,using immunohistochemistry envision method to detect of CD34 antigen and ki‐67 antigen ,and observing effect of Solanine group ,VEGF antibody group ,Solanine+ VEGF antibody group and the effect on the tumor angiogenesis and tumor proliferation .Results The tumor angiogenesis ,CD34 antigen and ki‐67 antigen of Sola‐nine+VEGF antibody group were significantly better than those of VEGF antibody group and Solanine group(P<0 .01);VEGF antibody group had statistical significant difference with Solanine group(P<0 .01);the effect of other three groups were better than that of the control group(P<0 .01) .Conclusion Solanine、VEGF antibody and Solanine combined with VEGF antibody could in‐hibit tumor angiogenesis and tumor proliferation of human colon cancer cell line HT‐29 to induce .It provides a new way for anti‐an‐giogenes .

Effects of Solanine on the human colon cancer in chicken embryo transplantation model angiogenesis / 重庆医学

Objective To establish chicken embryo transplantation model of human colon cancer and to research the effect of so‐lanine on angiogenesis .Methods Cases with chicken embryos were divided into the low‐,mid‐and high dose solanine group and con‐trol group ,with 10 cases in each groups ,and then the cultured human colon cancer cell line HT‐29 cell lines were inoculated to the chicken embryo villus allantois membrane (CAM ) .We observed the characteristics of the transplanted tumor in CAM angiogenesis by the stereo microscope .Image analysis software of Image‐pro plus 6 .0 and immunohistochemical method were used to observe the effect of different dose of solanine on angiogenesis .Results HT‐29 cell lines were inoculated to CAM 3-5 days ,a large number of blood vessels concentrated in tumors ,growing into or acrossing the surface of tumors .While tumors also rapidly growed .We took photo on the 5th day after receiving medicine and did imaging analysis .Then we calculated the area of angiogenesis in experimental group ,which was significantly lower than that of the control group ,quantitatively in a dose‐dependent manner .There were signifi‐cant differences among the groups(P<0 .01) .Microvascular density of 3 different dose of solanine was significantly lower than that of the control group by immunohistochemical method ;the expression of Ki‐67 antigen index decreased gradually ,which was highest in the control group ,and there were significant differences among the groups (P<0 .01) .Conclusion Solanine could inhibit angio‐genesis induced by human colon cancer HT‐29 cell lines obviously ,thus inhibiting the growth of tumor and providing an important basis for the treatment of anti‐tumor angiogenesis .

Solanine induces human breast cancer MCF-7 apoptosis through mitochondrial pathway / 中国药学杂志

OBJECTIVE: To study the effects of solanine on apoptosis of MCF-7 cells and explore the related mechanism.

Effects of solanine on microtubules system of human breast cancer MCF-7 cells / 中草药

Objective To investigate the effects of solanine on microtubular system in MCF-7 cell line. Methods Proliferation inhibition of MCF-7 cell line was evaluated by MTT assay. Cell cycle of MCF-7 cells was analyzed and the changes of a-tubulin protein and microtubule-associated protein 2 (MAP-2) protein were detected by flow cytometry. Results The IC50 of MCF-7 cells was 22.08 μg/mL. Solanine could induce MCF-7 cells arrested in S phaseand increase the levels of a-tubulin and MAP-2 in MCF-7 cell line. Conclusion Solanine could inhibit the MCF-7 cell proliferation by increasing a-tubulin and MAP-2 expression and inducing MCF-7 cells arrested in S phase.

Antihepatocarcinoma Effect of Solanine and Its Mechanisms / 中草药·英文版

ObjectiveTo explore the antitumor effect of solanine and its mechanisms.MethodsThe in vivo antitumor effect of solanine was observed using models developed through in vivo transplantation of tumor cells; In vitro lines of sensitive antitumor cells were selected from the digestive system using MTT assay; The effect of solanine on cell morphology was observed using transmission electronic microscopy; The morphology of apoptotic cells was observed using Annexin V/PI double staining and laser confocal scanning microscopy (LCSM); The rate of cell apoptosis was measured using Annexin V/PI double staining and flow cytometry; The concentration of intracellular Ca2+ ([Ca2+]1) was determined using Fluo-3/AM staining and LCSM; The membrane potential of cellular mitochondria was determined using TMRE staining and LCSM; The protein expression of Bcl-2 and Bax was measured using immunological marking and LCSM; And the activity of caspase-3 was measured using the colorimetric method.ResultsSolanine could inhibit the growth of tumor weight in S180 tumor-bearing mice and prolong the survival time of H22 tumor-bearing mice.MTT assay revealed that HepG2 cells were quite sensitive to solanine because solanine could induce morphological changes in HepG2 cells,with the rate of early apoptosis being 4％,8.5％,and 20.1％,for HepG2 cells treated for 24 h with solanine at concentration of 0.4,2,and 10 μg/mL,respectively.Solanine could raise the [Ca2+]i and lower the membrane potential.It could reduce the protein expression of Bcl-2 while increase that of Bax,thus increasing the activity of caspase-3.ConclusionThe obvious antitumor activity of sotanine in human hepatocarcinoma is demonstrated.This inhibitory effect is achieved through solanine decreasing the Bcl-2/Bax ratio,thus increasing [Ca2+]i,which could enhance the enzymatic activity of the caspase family,thus inducing the apoptosis of HepG2 cells.

Solanine on apoptosis by regulating glutathine hormone and reactive oxygen species to damage mitochondrial structure in HepG2 cell / 中草药

Objective To study the relationship between apoptosis and mitochondria damage.Methods The cell morphology was observed under inverted microscope.Staining with Annexin V/PI,the cell apoptosis was observed by laser confocal scan microscope and the apoptosis rate was analyzed by flow cytomety. Transmission electron microscope(TEM) was used to observe mitochondrial structure.ROS was determined by confocal.Colorimetry assay was adopted to determine the reduced glutathione hormone (GSH).Results The cell number in solanine groups was fewer than that in control group,and some died.Staining with Annexin V/PI,Annexin V-FITC was high fluorescent in solanine groups,which is obviously apoptosis features.The early apoptosis rate is 4.0%,8.5%,and 20.1%inducing by 0.4,2,and 10 mol/L solanine for 24 h,respectively.Swelling mitochondria,absence of mitochondria crests and vesicles were found in mitochondria of HepG2 cell treated by solanine for 24 h.The ROS levels increased and GSH level decreased in solanine groups.Conclusion Solanine could induce the apoptosis and the effect may be attributed to decrease GSH,which interrupts the immediate remove of ROS so as to damage the mitochondria.

Effect of solanine on DNA and RNA in tumor cell of tumor-bearing mice / 中草药

Objective To observe the effects of solanine on DNA and RNA level in tumor cell of S_(180) and H_(22) tumor-bearing mice.Methods S_(180) and H_(22) mice were divided into solanine(37.50、18.75,and(9.37) mg/kg) groups,negative control group and cytoxan(30 mg/kg) group,whom were given drugs by sc.Levels of RNA and DNA in tumor cell membrane in every groups were examined,respectively by laser scanning confocal microtechnic technology.Results Solanine(37.50、18.75 mg/kg) groups reduced the ratio of RNA and DNA in tumor cell of both S_(180) and H_(22) mice.Conclusion Solanine can reduce the ratio of RNA and DNA in tumor cell of S_(180) and H_(22) mice,which may be one of the mechanisms of solanine's(antitumor) effect.