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Objective:To study the mechanism of aloesin in inducing apoptosis in human non-small cell lung cancer (NSCLC) A549 cells, so as to inhibit its proliferation. Method:A549 cells in logarithmic growth phase were collected, and cell counting kit-8 (CCK-8) was used to detect the effect of different concentrations of aloesin (2, 4, 8, 16, 32, 64, 128 μmol·L-1) on the proliferation of A549. Effect of aloesin (0, 16 μmol·L-1) on the number of clones formed in A549 cells and the size of clone formation was determined by crystal violet staining. effect of aloesin on apoptosis of A549 cells was detected by annexin V/propidium iodide(PI)apoptosis kit staining. Hoechst staining was used to detect the phenomenon of apoptotic nuclear pyknosis. Western blot was used to detect aloesin's effect on death-related protein expressions of Bcl-xl/Bcl-2 associated death promoter (Bad), cleaved-Caspase-3,cl-Caspase-3(Asp175), Caspase-3, cleaved poly ADP-ribose polymerase (cl-PARP), poly ADP-ribose polymerase (PARP) in A549 cells. In vivo, 5-week-old nude mice were subcutaneously inoculated with 2×106 A549 cells, and randomly divided into the medication group and the blank group. aloesin or normal saline was intraperitoneally injected for 4 weeks, and the tumor volume of nude mice was measured weekly. The body weight of the mice was observed, and the appearance of the nude mice was observed. Result:Aloesin inhibited the proliferation and cloning of A549 cells in a concentration-dependent manner (PPPPPPin vivo, aloesin significantly shrank the volume of subcutaneous tumors in mice, reduced tumor weight, with a better appearance than that of the control group. Conclusion:Aloesin may inhibit the expression of NSCLC by inducing apoptosis of A549 cells, and is safe to use, with no inhibitory effect on the body weight of mice.
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Objective:To observe effect of Zeqi Tang in intervening mice with orthotopic lung cancer model, in order to observe its anti-tumor mechanism. Method:An in situ mouse model of non-small cell lung cancer was established through intrapulmonary injection with 1×105 LLC-luc cells. The model mice were intragastrically administered with Zeqi Tang(0.171 g·mL-1) or normal saline for 35 days. Appearance (spirit, hair, appetite, sleep), survival period and Zeqi Tang anti-tumor effect were observed, weekly vital imaging was performed to detect the fluorescence signal in the lungs of mice. Flow cytometry was used to detect the NK cell content in the spleen of the model mice. CD107α was used to detect the degranulation of NK cells in the spleen of mice after administration of Zeqi Tang. Kromasil 100 5 C18 column was used and eluted with acetonitrile-0.025%phosphoric acid in a gradient mode, with flow rate at 1.0 mL·min-1, column temperature at 35℃ and detection wavelength of 265 nm, as to establish the fingerprint of Zeqi Tang. The fingerprints of 10 batches of samples was evaluated by using the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System Software (2012 Edition) recommended by the Chinese Pharmacopoeia Commission, in order to complete the quality control of Zeqi Tang. Result:Zeqi Tang could significantly inhibit the lung fluorescence signal of lung cancer in situ model mice and prolong the survival of mice(PPPα also increased significantly(PConclusion:Zeqi Tang may enhance the tumor growth and prolong the survival period of mice by up-regulating the number of NK cells in mice and enhancing their degranulation function. The evaluation of similarity of HPLC fingerprint of Zeqi Tang reflects the quality of lacquer soup to a certain extent, and can provide reference for further study.
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Aim To investigate the effects of Bigelovii A on autophagy and its mechanism.Methods Fluorescence microscope,flow cytometry and Western blot were employed to analyze autophagy.Western blot was used to detect the protein expressions of mTOR pathway.MTT colorimetry was used to assay cell viability after treatment with 3-MA and Bigelovii A or Bigelovii A alone.Results Bigelovii A-treated MCF7 cells displayed a dramatic increase in the number of MDC-labeled vesicles and the expressions of LC3-Ⅱ,indicating cell autophagy.Ⅰt was proved that in MCF7 cells,Bigelovii A inhibited mTOR signaling by decreasing Akt and p-ERK.Consistently,Bigelovii A decreased phosphorylation levels of mTOR,p70S6K (Ser371,Thr389) and 4EBP1 proteins.Inhibiting Bigelovii Ainduced autophagy with the autophagy inhibitor 3-methyladenine significantly decreased cell viability,which suggested that Bigelovii A-induced autophagy played a pro-survival role.Conclusion Bigelovii A is likely to induce autophagy through inhibiting mTOR pathway.
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Aim To demonstrate that bigelovii E,one of the triterpenoid compounds isolated from Salicornia bigelovii Torr. ,induced apoptosis and inhibited prolif-eration of human breast cancer cells MCF-7. Methods MTT and Clonogenic were used to detect the anti-proliferative effect of bigelovii E on human tumor cells. Hoechst 33258 staining was used to observe the chan-ges of cell morphology in the treatment with bigelovii E. The effect of bigelovii E on cell apoptosis,cell mi-tochondrial membrane potential and the ROS level was analyzed by flow cytometry. The expression of mito-chondrial apoptotic pathway and its regulated proteins were analyzed by Western blot. Results Bigelovii E was most sensitive to MCF-7;bigelovii E had lower toxicity to normal cells HLF1;bigelovii E inhibited the colony formation of breast cancer MCF-7;the apoptosis pattern induced by bigelovii E was detected by Hoechst 33258;Western blot showed that bigelovii E could down-regulate the phosphorylation of mTOR,p70S6K and 4-EBP,up-regulate the expression of Bcl-xl and Mcl-1 proteins,promote the mitochondrial apoptosis pathway,reduce the mitochondrial membrane poten-tial,and induce ROS,leading to mitochondrial func-tion damage,and ultimately inducing apoptosis. Con-clusion Bigelovii E regulates mitochondrial apoptosis pathway through mTOR pathway,induces apoptosis and has a good anti-tumor effect.
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OBJECTIVE@#To investigate the effect of Chaiqinchengqi decoction (CQCQD) on serum amyloid A (SAA) in severe acute pancreatitis (SAP) patients.@*METHODS@#Thirty-five participants enrolled and were randomly assigned into either a treatment condition (n = 17, treated with CQCQD) or a control condition (n = 18, treated with placebo) 24 hours following the onset of the disease. No statistical difference was observed in either group at baseline. Upon admission, the Acute Physiology and Chronic Health Evaluation score II (APACHE II), SAA, serum C-reactive protein (CRP) and interleukin-6 (IL-6) were measured, as well as on the first, 3rd and 7th day and were compared between the two groups. Organ complications, infection, operation rate, mortality and hospital stay were also compared.@*RESULTS@#The duration of acute respiratory distress syndrome, acute hepatitis, acute renal failure, gastrointestinal failure and blood coagulation dysfunction were shorter in the treatment group than in those in the control group (P < 0.05). The secondary infection rates and the hospital fees in the treatment group were lower than those in the control group (P < 0.05) as well as length of hospital stay (P < 0.01). After 3 days of hospitalization, the APACHEII, score SAA levels, serum CRP and IL-6 in the treatment group was lower than those in the control group (P < 0.05). SAA was positively correlated with serum CRP (R = 0.346, P = 0.042), Ranson score (R = 0.442, P = 0.008) and serum IL-6 (R = 0.359, P = 0.034). The area under the receiver operating characteristic curve of admission SAA predict pancreatic necrosis (PN) was 0.815 (95% CI: 0.625-0.954; P = 0.006). The best cut-off value of admission SAA was 7.85 mg/L with the sensitivity 84.6% and specificity 68.2%.@*CONCLUSIONS@#The CQCQD can reduce the duration of organ damage through lowering the SAA in SAP patients and the SAA can early predict the PN and severity of SAP patients.