Angiotensin II ; metabolism ; Cisplatin ; adverse effects ; Humans ; Kidney ; drug effects ; Kidney Diseases ; chemically induced ; metabolism

Glutathione(GSH) has a very important role in detoxification of cells and is closely related to antitumor drug-resistance of cancer cells. In order to evaluate the importance of glutathione metabolism in the drug-resistant cancer cells, the concentration of celluar GSH and activities of y-glutamylcysteine synthetase(GCS), y-glutamyl transpeptidase (GGT) and glutathione S-transferases(GST) in the adriamycin, vincristine, or cisplatin resistant L1210 (L1210AdR; L1210VcR, or L12100s) sublines were measured. Expression and amplification of GCS, GGT, and GST-i7 genes were also observed in the parent L1210 and the drug-resistant L1210 sublines. The concentration of GSH was increased 5.34 fold in L12100s, 2.83 fold in L1210VcR, and 1.78 fo-d in L1210AdR, compared to L1210. The activities of GCS and GGT were -increased in drug-resistant L1210 sublines. The GST activity was increased in L1210VcR and L1210Cis but decreased in L1210AdR compared to L1210. Expression of GCS, GGT, and GST-rr genes were increased in the resistant L1210 sublines compare to the parent L1210 in northern blot analyses. Overexpression of GCS, GGT, and GST-77 were observed in the resistant sublines, and the increases of the concentration of glutathione and the activities of GCS and GGT in the resistant sublines may be involved in a part of the drug-resistance in the resistant sublines.
Blotting, Northern ; Cisplatin ; Doxorubicin ; Drug Resistance ; Glutathione* ; Humans ; Metabolism* ; Parents ; Vincristine

Glutathione (GSH) is a well-known antioxidative cellular component which is ubiquitous in nature. Several enzymes involved in GSH metabolism and recycling have been found to play important roles in detoxification of xenobiotics and free radicals. In this study, total GSH content, activity of GSH peroxidase and GSH reductase were measured in liver and kidney of cisplatin treated rats. Total GSH content (mM/g protein) of liver was higher in cisplatin treated rats (1.51±0.28) than of nontreated control (0.95±0.28), and in kidney, it was also higher in cisplatin treated rats (0.87±0.20) than that of control (0.68±0.14). The activity of GSH peroxidase (µM/mg protein/min) was lower in liver of cisplatin treated rats (348.0±18.54) than that of control (415.5±53.15), in kidney it was increase din cisplatin treated rats (380.5±51.86) compared to control (327.3±20.36). The activity of GSH reductase (µM/mg protein/min) was higher in liver of cisplatin treated rats (3.09±0.88) than that of control (2.28±0.61), in kidney it was also higher in cisplatin treated rats (8.50±2.62) than that of control (3.30±1.10). In summary, detoxification of ciplatin was revealed lesser effect in kidney as show increasion of GSH peroxidase and reductase and detoxification of cisplatin was expressed effectively in liver by increasing of GSH content and decreasing GSH peroxidase.
Animals ; Cisplatin* ; Free Radicals ; Glutathione* ; Kidney* ; Liver* ; Metabolism* ; Oxidoreductases ; Peroxidase ; Rats* ; Recycling ; Xenobiotics

OBJECTIVE: Chemotherapeutic drugs, such as cisplatin (CP), which are associated with oxidative stress and apoptosis, may adversely affect the reproductive system. This study tests whether administration of propolis and nano-propolis (NP) can alleviate oxidative stress and apoptosis in rats with testicular damage induced by CP. METHODS: In this study, polymeric nanoparticles including propolis were synthesized with a green sonication method and characterized using Fourier transform-infrared spectroscopy, Brunauer-Emmett-Teller, and wet scanning transmission electron microscopy techniques. In total, 56 rats were divided into the following seven groups: control, CP, propolis, NP-10, CP + propolis, CP + NP-10, and CP + NP-30. Propolis (100 mg/kg), NP-10 (10 mg/kg), and NP-30 (30 mg/kg) treatments were administered by gavage daily for 21 d, and CP (3 mg/kg) was administered intraperitoneally in a single dose. After the experiment, oxidative stress parameters, namely, malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GPx), and catalase (CAT), and apoptotic pathways including B cell leukemia/lymphoma-2 protein (Bcl-2) and Bcl-2-associated X protein (Bax) were measured in testicular tissues. Furthermore, sperm quality and weights of the testis, epididymis, right cauda epididymis, seminal vesicles and prostate were evaluated. RESULTS: Propolis and NP (especially NP-30) were able to preserve oxidative balance (decreased MDA levels and increased GSH, CAT, and GPx activities) and activate apoptotic pathways (decreased Bax and increased Bcl-2) in the testes of CP-treated rats. Sperm motility in the control, CP, and CP + NP-30 groups were 60%, 48.75%, and 78%, respectively (P < 0.001). Especially, NP-30 application completely corrected the deterioration in sperm features induced by CP. CONCLUSION The results show that propolis and NP treatments mitigated the side effects of CP on spermatogenic activity, antioxidant situation, and apoptosis in rats.
Animals ; Antioxidants/metabolism* ; Cisplatin/toxicity* ; Male ; Oxidative Stress ; Propolis ; Rats ; Rats, Sprague-Dawley ; Sperm Motility ; Testis

OBJECTIVE: To investigate the effect of connexin43 (Cx43) protein on autophagy in cisplatin (DDP)-resistant testicular cancer I-10 cells. METHODS: The expression of Cx43 proteins in testicular cancer I-10 cells and I-10/DDP cells were detected with Western blotting. I-10/DDP cells were transfected with a full- length mouse Cx43 vector (mCx43) Lipofectamine, the empty vector or Lipofectamine (blank control group), and the changes in the expressions of LC3 and p62 proteins were determined with Western blotting. mCherry-GFP-LC3B transfection and transmission electron microscopy were used to analyze the changes in autophagy of the cells with Cx43 overexpression. RESULTS: Cx43 was significantly decreased in I-10/DDP cells compared with I-10 cells ( < 0.01). Transfection of the I-10/DDP cells with mCx43 vector resulted in significantly increased Cx43 expression in the cells ( < 0.01) and caused significantly decreased expression of LC3-Ⅱ ( < 0.01) and increased expression of p62 ( < 0.05) as compared with the negative control cells. Both transmission electron microscopy and mCherry-GFP-LC3B transfection showed that the number of autophagosomes was obviously reduced in mCx43-transfected cells as compared with the negative control cells. CONCLUSIONS Cx43 inhibits autophagy in cisplatin-resistant testicular cancer I-10 /DDP cells.
Animals ; Autophagy ; Cell Line, Tumor ; Cisplatin ; Connexin 43 ; metabolism ; Drug Resistance, Neoplasm ; Male ; Mice ; Testicular Neoplasms ; metabolism ; pathology

OBJECTIVE: To study the effect of forsythiaside on the expression of c-jun induced by cisplatin in the cochlea of guinea pig. METHOD: Thirty guinea pigs were randomly divided into control group (10), cisplatin group (10) and forsythiaside group (10). The ototoxicity model was done with intraperitoneal injection of cisplatin solution (8 mg/kg per day) for 7 days. Forsythiaside (25 mg/kg per day) was injected 30 min before cisplatin solution treated in guinea pigs of forsythiaside group for 7 consecutive days. The saline instead of cisplatin was injected in normal control group. The distortion product otoacoustic emission (DPOAE) was detected before animals were killed. The expression of c-jun in cochlea of guinea pigs was detected by western blotting. The expression of c-jun mRNA in cochlea of guinea pigs was detected by reverse transcriptase polymerase chain reaction (RT-PCR). RESULT: DPOAE amplitudes in cisplatin group was significantly lower than in control group (P < 0.01). Compared with cisplatin group, DPOAE amplitudes in forsythiaside group was increased significantly (P < 0.05). The expression of c-jun protein and mRNA were significantly increased in cisplatin group than in control group (P < 0.01). Compared with cisplatin group, the expression of c-jun protein and mRNA were significantly decreased in forsythiaside group. CONCLUSION Forsythiaside can significantly reduce the side effects induced by cisplatin through down-regulating the expression of c-jun.
Animals ; Cisplatin ; toxicity ; Cochlea ; drug effects ; metabolism ; Female ; Glycosides ; pharmacology ; Guinea Pigs ; Male ; Proto-Oncogene Proteins c-jun ; metabolism

Extracellular matrix (ECM) has been implicated in tumor progress and chemosensitivity. Ovarian cancer brings a great threat to the health of women with a significant feature of high mortality and poor prognosis. However, the potential significance of matrix stiffness in the pattern of long non-coding RNAs (lncRNAs) expression and ovarian cancer drug sensitivity is still largely unkown. Here, based on RNA-seq data of ovarian cancer cell cultured on substrates with different stiffness, we found that a great amount of lncRNAs were upregulated in stiff group, whereas SNHG8 was significantly downregulated, which was further verified in ovarian cancer cells cultured on polydimethylsiloxane (PDMS) hydrogel. Knockdown of SNHG8 led to an impaired efficiency of homologous repair, and decreased cellular sensitivity to both etoposide and cisplatin. Meanwhile, the results of the GEPIA analysis indicated that the expression of SNHG8 was significantly decreased in ovarian cancer tissues, which was negatively correlated with the overall survival of patients with ovarian cancer. In conclusion, matrix stiffening related lncRNA SNHG8 is closely related to chemosensitivity and prognosis of ovarian cancer, which might be a novel molecular marker for chemotherapy drug instruction and prognosis prediction.
Female ; Humans ; Cisplatin/pharmacology* ; Elasticity/physiology* ; Etoposide ; Extracellular Matrix/physiology* ; Ovarian Neoplasms/metabolism* ; RNA, Long Noncoding/metabolism*

OBJECTIVETo investigate the effects of high-affinity tyrosine kinase receptors TrkB, TrkC and the low-affinity neurotrophin receptor p75 in spiral ganglion cell (SGC) of cisplatin-induced ototoxicity.

METHODSThe 50 adult Wistar rats were divided randomly into 5 groups received intraperitoneal injection of cisplatin with vary dose. Control group was received equivalent volumes of saline. The group received 1 day intraperitoneal injection was cisplatin treated at a dose of 5 mg/kg and killed at next day. The group received 3 days was cisplatin treated for 3 days at same dose daily and then killed at next day. The group A received 5 days was cisplatin treated for 5 days and killed at next day. The group B received 5 days was cisplatin treated for 5 days and then were sacrificed after 7 days. The change of mRNA level of neurotrophin receptors in cochlear tissue were examined by RT-PCR. The expressing pattern of TrkB, TrkC, P75 in damaged cochlea were study by immunochemistry using antibodies against TrkB, TrkC, P75 protein.

RESULTSThe research data showed the expression of Trk B, Trk C, p75 exhibited in SGC was dynamic along with the administration lasting. The mRNA and protein level of Trk B (x(-) +/- s) at day 1 and 3 after cisplatin treatment were 0.76 +/- 0.06, 88.78 +/- 4.28, 0.82 +/- 0.09 and 91.64 +/- 4.06, with significant difference among those and other groups (P < 0.05). The mRNA and protein level of TrkC at day 1 after cisplatin treatment were 0.80 +/- 0.06 and 89.66 +/- 2.76, with significant difference among that and other groups (P < 0.05). The mRNA and protein level of p75 at the control group and cisplatin treated groups were 0.64 +/- 0.04, 55.16 +/- 3.10, 0.77 +/- 0.04, 78.46 +/- 3.86, 1.01 +/- 0.09, 105.02 +/- 6.61, 1.18 +/- 0.09, 111.10 +/- 6.08, 0.51 +/- 0.04 and 42.74 +/- 5.20, with significant difference among the control group and cisplatin treated groups (P < 0.05).

CONCLUSIONSThe expression of Trk B increased to peak at day 1 - 3 after cisplatin treatment and decreased at day 5 early and following weeks. The expression of Trk C went up to peak at day 1 after cisplatin treatment and went down during subsequently time. P75 kept a trend of continuance increased during the drug treatment and decrease at drug stopped. The expression of Trk B, Trk C and P75 may be involved in cochlear insult with cisplatin-induced. Trk B and Trk C may play an important role in the reparative process of cochlear, especially at early stage of the damage. P75 could promote SGC apoptosis in cisplatin-induced neurotoxicity.

Animals ; Cisplatin ; toxicity ; Male ; Rats ; Rats, Wistar ; Receptor, Nerve Growth Factor ; metabolism ; Receptor, trkB ; metabolism ; Receptor, trkC ; metabolism ; Spiral Ganglion ; drug effects ; metabolism

OBJECTIVETo explore the changes and significance of the level of reactive oxygen species (ROS) and mitochondria membrane potential (Delta Psi) in HepG2 cells under the stress of cisplatin (CDDP).

METHODSHepG2 cells were incubated with CDDP. The changes in the level of ROS were determined by a probe (2,7-dichloro fluorescein-ciactate, DCFH-DA) and the changes of Delta Psi were reflected as changes of intensities of fluorescence seen under a laser scan microscope using a probe (rhodamine-123). All these changes in cells at 0 h, 24 h, 48 h, 72 h, 120 h, 168 h were dynamically observed.

RESULTSThe level of ROS was much higher after the CDDP treatment than the non-treated, and the increase lasted for 24 h and 48 h. Then it started to decrease at 72 h, gradually returning to normal level at 120 h. Under the selective pressure of CDDP, the fluorescence intensity of rhodamine-123 in HepG2 cells was decreasing at 24 h and 48 h, then gradually started to increase at 72 h. There were no such changes in the cells of the controls.

CONCLUSIONThe changes of ROS and Delta Psi in HepG2 cells under the pressure of CDDP suggest that the cells change themselves adapting to such pressures.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cisplatin ; pharmacology ; Hepatocytes ; cytology ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Membrane Potentials ; drug effects ; Mitochondria, Liver ; physiology ; Reactive Oxygen Species ; metabolism ; Tumor Cells, Cultured

OBJECTIVE: To investigate the effectiveness of cisplatin on the expressions of Bcl-2 and Bax in cochlea and spiral ganglion cells (SGC) of guinea pigs. METHOD: Twenty guinea pigs were randomly divided into cisplatin (n = 10) and control groups (n = 10). Cisplatin group were administrated with a dose of intraperitoneal injection of 16 mg/kg, while the control group were received intraperitoneal injection of normal saline as placebo. Before and 7 days following injections, the ototoxic effect was measured with distortion product otoacoustic emission (DPOAE). Bcl-2, Bax in cochlea were detected by Western Blot. Immunohistochemical staining was used to detect the protein levels of Bcl-2 and Bax in spiral ganglion cells. RESULT: In control and cisplatin group, Bcl-2 protein levels were 0.727 8 ± 0.016 9 and 0.467 6 ± 0.020 1, Bax protein levels were 0.384 8 ± 0. 0217 and 0.735 6 ± 0.022 3 in cochlea respectively, both P < 0.01. In Control and cisplatin group, the grey values of Bcl-2 in SGC were 99.00 ± 2.42 and 149.80 ± 2.37 respectively, the grey values of Bax were 154.50 ± 2.80 and 104.50 ± 3.09 respectively, both P < 0.05. CONCLUSION Decreased expression of Bcl-2 and increased expression of Bax may be involved in cisplatin-induced apoptosis in cochlea and SGC of guinea pigs.
Animals ; Apoptosis ; Cisplatin ; pharmacology ; Cochlea ; metabolism ; Guinea Pigs ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Spiral Ganglion ; drug effects ; metabolism ; bcl-2-Associated X Protein ; metabolism