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AIM: To investigate the role of ribonucleotide reductase M2 (RRM2) in 5-fluorouracil (5-FU) resistance of HCC cells and to develop potential strategies to enhance the sensitivity of HCC cells to 5-FU. METHODS: The expression of RRM2 was examined by Western blot in BEL/5-FU cells and BEL7402 cells. The expression of RRM2 was down-regulated through RNA interference (RNAi) technology and the activity of RRM2 was inhibited by the RRM2 inhibitor 3-AP. The cell proliferation ability was detected by CCK-8 assay and colony formation assay, and the apoptosis was analyzed by Confocal high-content system. RESULTS: The expression level of RRM2 was increased by 2.5-fold in BEL/5-FU cells compared with BEL7402 cells. Compared with control siRNA, the half maximum inhibitory concentration IC
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Objective To discuss the protective effects of thioltransferase (TTase) on oxidative damaged human lens epithelial cells (HLEC) induced by ultraviolet radiation.Methods HLEC were cultured in vitro and then randomly divided into 4 groups:Normal group:normal cultured HLEC;UV group:normal cultured HLEC + UV radiation (with 302 nm UV radiation irradiation intensity 55.56 μW · cm-2 for 15 minutes,totaling irradiation volume 500 J · m-2);TTase siRNA group:HLEC transfected with TTase siRNA;TTase siRNA + UV group:HLEC transfected with TTase siRNA + UV radiation(with 302 nm UV radiation irradiation intensity 55.56 μW · cm-2 for 15 minutes,totaling irradiation volume 500 J · m-2).TTase mRNA expression was measured by qRT-PCR,the cell proliferation was detected by LDH Assay Kit,and the TTase activity was measured.TTase expression was detected by Western blotting.The levels of TGSH,GSH and GSSG of HLEC were measured,and then GSSG/T-GSH ratio was calculated.Results Cell proliferation ability in UV group,TTase siRNA group and TTase siRNA + UV group were decreased by 21.0%,17.0% and 29.0% compared with normal group (all P < 0.05).TTase activity in UV group was 2.1 times of the normal group,TTlase siRNA group was 67.0% of the normal group,Tlase siRNA + UV group was 1.3 times of TTase siRNA group (all P < 0.05).TTase expression in UV group was 3.9 times of the normal group,TTase siRNA group was 35.0% of the normal group,TTase siRNA + UV group was 3.0 times of siRNA group (all P < 0.05).GSH content in UV group,TTase siRNA group and TTase siRNA + UV group were 68.4%,79.0%,61.7% of the normal group (all P < 0.05).GSSG content in UV group,TTase siRNA group and TTase siRNA + UV group were 2.3 times,1.4 times,3.7 times of the normal group (all P < 0.05).GSSG/T-GSH in UV group,TTase siRNA group and TTase siRNA + UV group were 3.1 times,1.7 times,5.2 times of the normal group (all P < 0.05).Conclusion TTase plays an important protective role in oxidative damaged HLEC induced by ultraviolet radiation.
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OBJECTIVE To observe and compare the cytotoxicity induced by andrographolide (AD)and its water soluble derivatives:andrographolide sodium bisulfite(ASB),active pharmaceutical ingredients of Chuanhuning and Yanhuning on human renal tubular epithelial cells (HK-2),and to explore the ASB-induced endoplasmic reticulum stress(ERS)mechanism. METHODS HK-2 cells were treated with the above four drugs respectively. The survival rate was examined by methyl thiazolyltetrazolium (MTT) assay and 50% inhibitory concentration (IC50) was calculated. In ASB treated group, Hoechst33342 staining and flow cytometry analysis were used to determine cell apoptosis, intracellular superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were examined, and the protein expressions of binding immunoglobulin protein (Bip),C/EBP-homologous protein (CHOP)and cysteine-containing aspartate-specific protease 4(caspase 4)were detected by Western blotting. RESULTS The four drugs inhibited HK-2 cell growth in a time-dependent and concentration-dependent manner. At 24 h,the IC50 of AD (30.6 μmol · L- 1) was lower than that of others. Active pharmaceutical ingredients of Chuanhuning and Yanhuning (16.2 and 15.6 mmol · L- 1) were very close,ASB was 29.4 mmol · L-1. ASB(0,15,30 and 60 mmol · L-1)increased the apoptotic rate and caused the decrease in SOD activity and the increase in MDA content in a dose-dependent manner. Compared with control group,the protein expression of CHOP increased (P<0.01) at 8 h with ASB (30 and 60 mmol · L-1)treatment,Bip and caspase 4 had no significant change. In addition,at 24 h, ASB(60 mmol·L-1) decreased the expression of Bip(P<0.05),ASB(30 and 60 mmol·L-1)promoted the expression of CHOP(P<0.01),and the protein expression of activated caspase 4 increased in a concentration-dependent manner(P<0.01). CONCLUSION AD and its water soluble derivatives have a toxic effect on HK-2 cells. CHOP and caspase 4 pathway related to ERS is involved in ASB-induced apoptosis.
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<p><b>OBJECTIVE</b>To investigate the inhibitory effects of phytosterols on abacterial prostatitis and discuss the possible mechanism.</p><p><b>METHOD</b>Xiaozhiling-induced chronic prostatitis model were used to observe the inhibitory effect of phytosterols on abacterial prostatitis. The changes of serum IL-2, IL-1beta and TNF-alpha were evaluated by enzyme-linked immunosorbent assay (ELISA). The expression of COX-2 and 5-LOX were evaluated by Western blot and immunohistochemistry.</p><p><b>RESULT</b>Treated by phytosterols (150 mg x kg(-1)), the number of white blood cells in xiaozhiling-induced chronic abacterial prostatitis rats was obviously decreased, the density of lecithin corpuscle in prostatic secretion increased and closed to control group. The edema, inflammatory infiltration of prostate were partly recovered compared with model group. The proliferation of chronic prostatitis were obviously decreased in phytosterols groups compared with model group in histological sections. Phytosterols could obviously reduce the serum IL-1beta, TNF-alpha, prostate COX-2 and 5-LOX expression and improve IL-2 level.</p><p><b>CONCLUSION</b>These results demonstrated that phytosterols had good therapeutic effects on chronic abacterial prostatitis. Participation of immune regulation and inhibiting COX-2 and 5-LOX expression may be the mechanisms of action.</p>
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Animais , Humanos , Masculino , Ratos , Doença Crônica , Terapêutica , Modelos Animais de Doenças , Interleucina-1beta , Sangue , Alergia e Imunologia , Interleucina-2 , Sangue , Alergia e Imunologia , Fitosteróis , Usos Terapêuticos , Extratos Vegetais , Usos Terapêuticos , Prostatite , Tratamento Farmacológico , Alergia e Imunologia , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa , Sangue , Alergia e ImunologiaRESUMO
<p><b>OBJECTIVE</b>To observe the effects of Qingkailing injection on RBL-2H3 cell degranulation and histamine release, and discuss the possible mechanism of anaphylactoid reaction induced by Qingkailing injection.</p><p><b>METHOD</b>RBL-2H3 cells were incubated with Qingkailing injection for 30 min. Then the morphological changes of cells were observed by transmission electron microscopy. Cell degranulation rate was detected by Alcian blue dye assay, Annexin V binding assay and beta-hexosaminidase assay, and cell histamine release rate was detected by ELISA.</p><p><b>RESULT</b>Different concentration of Qingkailing injection can induce the typical morphological changes in RBL-2H3 cell with degranulation. The rates of degranulation and histamine release in Qingkailing injection treated cells were significantly increased and dose-dependent.</p><p><b>CONCLUSION</b>RBL-2H3 cell degranulation and histamine release can be induced by single administration of Qingkailing injection, and then induced anaphylactoid reaction, which may be one of the possible mechanisms of serious adverse induced by Qingkailing injection for the first administration in clinic.</p>
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Animais , Ratos , Basófilos , Alergia e Imunologia , Fisiologia , Degranulação Celular , Medicamentos de Ervas Chinesas , Farmacologia , Histamina , MetabolismoRESUMO
Objective To investigate the effect of astragalus mongholicus injection in the treatment of viral myocarditis(VM).Methods 80 VM patients were divided into 2 groups randomly and equally.Patients in the control group received routine treatment and patients in the observe group were given astragalus mongholicus injection besides the routine treatment.After 1 month,cardiac muscle enzymes and the clinical effects of the 2 groups were analyzed and evaluated.Results After 1 month,the effective power of the observe group was significantly higher than that of control group( P< 0.05 ).The incidence of chest distress,thoracodynia and cordis in the observe group also lower than that of control group.AST,CPK and LDH in the observe group decreased significantly after treatment(P<0.05 ).But we found no changes in the control group.Conclusion The clinical effects of the VM treatment can be improved by using astragalus mongholicus injection.
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AIM To explore the inhibition in in vitro p roliferation human prostate cancer PC 3 cells induced by melatonin and the effe ct of combination of melatonin and 5-Fu. METHODS Cell growth cu rve, MTT assay, and acid phosphatase(ACP)activity were used to determine cell proliferation. Coomassie brillient blue assay and modified diphenylamine assay w ere used to measure the content of protein and DNA in PC 3 cells. ACP activity was measured by modified phenyl phosphat e method of kind and king and modified method of lowry. Co-administration effec t of MLT and 5-fluorouracil(5-Fu)on PC 3 cell proliferation was determined by MTT assay. RESULTS The IC 50 of MLT for inhibiting PC 3, CoLo205, K562, and HeLa cell proliferation was(1 20?0 30),(1 27?0 12) ,(1 60?0 16)and(2 25?0 11)mmol?L -1 , respectively. And MLT inhib ited all above cells proliferation in a concentration-dependent manner. The protein and DNA content and ACP activity in PC 3 cell treated with 2 mmol?L -1 MLT for 96 h were redu ced 58 77%, 54 97% and 89 24%, respectively. When PC 3 cells were treated wi th MLT in combination with 5-Fu 0 5 mg?L -1 , the inhibition rate enhance d. CONCLUSION MLT inhibited PC 3, CoLo205, K562, and HeLa cell p roliferation, and reduced the content of protein and DNA and ACP activity in PC 3 cell. MLT and 5-Fu co-administration enhanced the inhibition rate of PC 3.er.TheproteinandDNAcontentandACPactivityinPC3 celltreatedwith 2mmol?L-1MLTfor 96hwerereduced5 8 77% ,5 4 97%and 89 2 4% ,respectively .WhenPC3 cellsweretreatedwithMLTincombinationwith5 Fu 0 5mg?L-1,theinhibitionrateenhanced .CONCLUSION MLTinhibitedPC3 ,CoLo2 0 5 ,K5 62 ,
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A number of compounds extracted from plants have cert ain physiological and pharmacological activity.Some of these compounds had been proved to have the ability to induce tumor cell apoptosis and demonstrated antitumor activity depending on their chemical structures.This review focuses on recent progress in the studies about the mechanism of tumor cell apoptosis induced by plant extracts including alkaloids,terpenes and lignans.
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Aim Organotin compounds are important organometallic chemicals with a variety of technical applications.Of these compounds,the trialkyltins,especially tributyltin,triphenyltin and trimethyltin,have been proved to have the distinct ability to induce tumor cell apoptosis and significant antitumor activity.They can induce tumor cell apoptosis through Fas receptor,mitochondrial,Ca 2+,MAPKs,p53,NF-?B and caspase/p38/ROS pathway.This paper reviewed recent progress in the studies on signal transduction in tumor cell apoptosis induced by trialkyltin compounds.
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Aim To study the effect of novel AChE inhibitors, 2-phenoxy-indan-1-one derivatives (YKY-1~7), against glutamatic acid-induced neurotoxicity in PC12 cells and on learning & memory impairment in dementia model mice induced by A?25~35 icv Methods The PC12 cells were preincubated with different concentrations of YKY-1~7 for 24 h and subsequently treated by glutamatic acid, at the high concentration of 2 mmol?L-1 for 15 min to induce cytotoxicity. The cell viability was assessed with MTT method.. Dementia model mice were made by intracerebroventricular injection (icv) of aggregated A?25~35. From the next day, the model mice were administered YKY-7 (2.5, 5, 10 mg?kg-1, ig) for 10 consecutive days and sham control mice or A? model control mice received daily ig saline. After the final treatment, the passive avoidance learning was tested, regional cerebral blood flow at cerebral cortex was assessed, and the activity of AChE in the cerebral cortex, hippocampus and blood serum were determined. Results Six out of the seven YKY compounds appeared to be effective against glutamatic acid-induced neurotoxicity in PC12 cells, with YKY-7 demonstrating the most activity. YKY-7 significantly ameliorated the learning and memory ability in dementia model mice induced by A?25-35 icv, slightly and selectively inhibited the cortical and hippocampal AChE, and gently increased the blood flow at cerebral cortex. Conclusion Some of 2-phenoxy-indan-1-one derivatives reported here have protective effects against glutamatic acid induced neurotoxicity in PC12 cells, and improve the learning and memory impairment induced by A?25-35, which may be partly attributable to its selective inhibition of AChE activity in the cerebral cortex and hippocampus.