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AIM: To study the correlation between the expression of recepteur d'origine nantais (RON) protein and CXC chemokine motif receptor 4 (CXCR4) protein and abiraterone resistance in patients with castration-resistant prostate cancer (CRPC). METHODS: From January 2017 to February 2020, 127 patients with CRPC who were treated with abiraterone in our hospital were selected. According to the status of drug resistance, they were divided into observation group (n=32, patients with abiraterone resistance) and control group (n=95, patients in remission). Immunohistochemistry and Western blot analysis were used to compare the expression of RON and CXCR4 protein between the two groups. Logistic regression analysis was used to conduct single-factor and multi-factor analysis of RON, CXCR4 protein and drug resistance, and receiver operating characteristic curve (ROC) and area under ROC (AUC) were used to analyze the value of RON and CXCR4 protein in predicting drug resistance. In addition, RON and CXCR4 inhibitors were added to abiraterone-resistant cell lines. The effects of the two on the apoptosis indicators of abiraterone resistance[caspase-3, caspase-9, apoptosis rate] were observed. RESULTS: Immunohistochemistry showed that the positive expression rate of RON in the observation group (71.88%, 23/32) was higher than that of the control group (27.37%, 26/95). The positive expression rate of CXCR4 in the observation group (65.63%, 21/32) was higher than that of the control group (12.63%, 12/95). Western blot detection showed that the RON and CXCR4 proteins in the observation group were higher than those in the control group (P 4.11, the sensitivity was 84.37%, and the specificity was 61.05% (P 3.42, the sensitivity was 75.00%, and the specificity was 80.00% (P< 0.05). The AUC of RON+CXCR4 protein predicting abiraterone resistance was 0.884 (95%CI: 0.815-0.934), the sensitivity was 87.50%, and the specificity was 83.16% (P< 0.05). CONCLUSION: The expression of RON and CXCR4 protein in CRPC patients increases significantly, which is closely related to the resistance of Abiraterone in patients and is expected to become a marker for predicting drug resistance. Inhibiting the expression of RON and CXCR4 proteins can promote the apoptosis of CRPC abiraterone resistant cells.
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BACKGROUND: Non-healing of refractory ulcer wounds is found to be associated with imbalance of apoptosis and dysregulation of inflammation. Previous studies have sampled a certain type of tissue in different individuals, but individual differences maybe exist among the study data. There is still no report on the correlation between apoptotic rate, expression levels of Bcl-2, Bax, Bax/Bcl-2 and tumor necrosis factor α (TNF-α) in wound tissue of diabetic foot ulcer with different disease courses. OBJECTIVE: To study the relationship between apoptotic rate and expression levels of Bcl-2, Bax and TNF-α in the wound tissue of diabetic foot ulcer with different disease courses and the mechanism of action. METHODS: Fifteen patients with typical diabetic foot of Wagner grades 2-3 were selected from the Department of Emergency, Chongqing Medical University. The wound tissue was classified as necrotic, transitional and normal tissue using the "4C” method after infection control. The apoptotic rate and levels of Bcl-2, Bax, TNF-α and Bax/Bcl-2 were detected using immunohistochemistry, and the results were statistically analyzed. The study protocol was approved by the Ethics Committee of the Second Affiliated Hospital of Chongqing Medical University, with the approval No. (2019)329. All patients signed an informed consent. RESULTS AND CONCLUSION: The apoptotic rates of normal, transitional and necrotic tissues of diabetic foot were significantly different, which were (16.67±2.48)%, (43.68±2.22)% and (72.12±4.53)%, respectively (P transitional tissue > normal tissue. There was a curve relationship between Bcl-2, Bax, Bax/Bcl-2, TNF-α expression and apoptotic rate. The curvilinear regression analysis indicated that the expression of TNF-α had a highly positive correlation with the expression of Bax and Bax/Bcl-2 and had a highly negative correlation with the expression of Bcl-2. Therefore, apoptotic mechanism and inflammatory reaction are involved in the pathological process of diabetic food ulcer, and the mechanism may be related to the down-regulation of Bcl-2 expression, up-regulation of Bax and TNF-α expression, and increase of Bax/Bcl-2, thereby increasing the apoptotic rate.
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OBJECTIVE： To study the effects of aspirin on the growth and autoghagy of human gastric cancer cells SGC-7901 and BGC-823. METHODS： SGC-7901 and BGC-823 cells were selected as research objects， with phosphate buffer （PBS） as negative control treated for 48 h， MTT assay was used to detect the effects of 1， 2， 4， 6， 8， 10 mmol/L aspirin， 5 mmol/L aspirin alone or combined with 2.5 μmol/L chloroquine， 2.5 μmol/L 3-methyladenine （3-MA） on survival rate of gastric cancer cells. Flow cytometry was used to detect the effects of 2 and 5 mmol/L aspirin， 5 mmol/L aspirin alone or combined with 2.5 μmol/L chloroquine and 2.5 μmol/L 3-MA on the apoptosis rate and cell cycle distribution of gastric cancer cells. Hoechst33258 staining was used to observe the effects of 5 mmol/L aspirin on morphology of gastric cancer cell nucleus； Transwell chamber test was adopted to detect the effects of 5 mmol/L aspirin on the migration of gastric cancer cell. Laser confocal scanning microscopy was used to observe the effects of 5 mmol/L aspirin on autophagy formation in gastric cancer cells. Western blot method was used to detect the effects of 2 and 5 mmol/L aspirin on the protein expression of autophagy markers LC3-Ⅱin gastric cancer cells. RESULTS： Compared with negative control group， aspirin could inhibit the survival rates of SGC-7901 and BGC-823 cells in dose-dependent manner， but had no significant effects on apoptosis rate of SGC-7901 and BGC-823 cells； SGC-7901 and BGC-823 cells were blocked in G1 phase. Compared with aspirin alone group， the survival rates of SGC-7901 and BGC-823 were increased significantly after treated with aspirin+chloroquine and aspirin+3-MA， while the distribution rate of SGC-7901 and BGC-823 cells at G1 phase were decreased significantly， with statistical significance （P＜0.05 or P＜0.01）. Compared with negative control group， there were no obvious DNA fragmentation fragments， apoptotic bodies and fragments of dense bright blue， while the number of migration cells were decreased significantly in SGC-7901 and BGC-823 cells after treated with aspirin （P＜0.001）； the number of autophagosome was increased significantly and the protein expression of LC3-Ⅱ was enhanced significantly （P＜0.05）. CONCLUSIONS： Aspirin can significantly inhibit the growth of SGC-7901 and BGC-823 cells， and arrest cell cycle in G1 phase， the mechanism of which may be associated with the activation of autophagy.
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Objective To investigate the effects of bevacizumab on cell morphology,apoptosis rate and apoptosis-related factors in human retinal pigment epitheliumcells.Methods Human retinal pigment epithelial cells (ARPE-19) were cultured in DMEM/F1 2 medium containing 0.25 g · L-1 bevacizumab and divided into 1-week dosing group and 2-week dosing group according to the different incubation time,respectively.Meanwhile,the control group was set up.Then,the cell morphology and apoptosis of each group were observed by light microscope and flow cytometry,accordingly.The expression levels of apoptosis-promoting genes P53,TP53INP1,Bax and apoptosisinducible gene Bcl-2 mRNA and protein were detected by RT-PCR and Western blot,respectively.Results Light microscopic observation revealed that the cells in the 1-week control group and the 2-week control group showed typical epithelial cell morphology,and were in stone-like,single-layer adherent-like growth.Compared with the control group,part of the cell morphology after bevacizumab treatment changed slightly,and the cells became rounded,the cell body was elongated and the boundary was poor.The apoptotic rates of the 1-week control group,l-week dosing group,2-weekcontrol group and 2-week dosing group were (5.57 ± 1.46) ％,(6.39 ± 1.25) ％,(6.88 ± 1.10)％ and (13.34 ± 1.94)％,respectively,and there was no significant difference in the apoptotic rate between 1-week control group,1-week dosing group and 2-week control group (both P ＞ 0.05),but the apoptotic rate of the 2-week dosing group was significantly higher than that of the 2-week control group,the 1-week dosing group and 1-week control group,and the differences were statistically significant (all P ＜0.01).RT-PCR and Western blot results showed that compared with the 1-week control group,the expression of P53,TP53INP1 and Bax mRNA was up-regulated in 1-week dosing group,but the expression of Bcl-2 mRNA was down-regulated,and the differences were statistically significant (all P ＜ 0.05).The expression levels of P53,TP53INP1 and Bax mRNA in 2-week dosing group were higher than those in 2-week control group and 1-week dosing group,while Bcl-2 mRNA expression was down-regulated,and the differences were statistically significant (all P ＜ 0.05).There was no significant difference in the expression of the above factors in 1-week control group and 2-week control group(all P ＞ 0.05).The protein expressions of above factors were similar to their mRNA expression.Conclusion Bevacizumab can alter the morphology of ARPE-19 ceils,increase the apoptotic rate and up-regulate the expression of apoptosis-promoting factors,but down-regulate the expression of apoptosis-suppression factors in ARPE-19 cells,which may be the reason for the loss of RPE layer in the macula after anti-VEGF therapy.
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This study was aimed to observe the protective effects of components of water extract from Qi-Xue Bing-Zhi recipe (CWQB) on myocardial cells from hypoxia/reoxygenation (H/R) injury,by optimizing the metabolism of highenergy phosphates and then preventing cell damage and early apoptosis correlatively.Myocardial cells were separated and extracted from newborn SD rats.And then,H/R models were made by depriving oxygen for 3 hours and then regaining for 2 hours.Cardiomyocytes were divided into four groups,which were the control (normal oxygen) group,H/R (H/R model) group,TMZ (H/R model + 100 μmol/L Trimetazidine,TMZ) group,and CWQB (H/R model + 1 mmol/L CWQB) group.High-performance liquid chromatography (HPLC) was used in the content determination of adenosine monophosphate (AMP),adenosine diphosphate (ADP),adenosine triphosphate (ATP) in each group for the calculation of total adenylic acid (TAN) and energy charge (EC).Enzyme linked immunosorbent assay (ELISA) was used in the detection of myocardial damage markers,such as creatine kinase-MB (CK-MB) and cardiac troponin T (cTnT).The early apoptosis of myocardial cells were assessed by flow cytometry (FCM).The results showed that compared with H/R group,contents of AMP in the TMZ group and CWQB group were decreased,while contents of ADP,ATP,TAN and EC were all increased in both groups.The increasing degrees of ADP,ATP and TAN in TMZ group were higher than those of the CWQB group (P ＜ 0.05).Contents of CK-MB and cTnT were significantly decreased in the TMZ group and the CWQB group.Content of cTnT decreased more significantly in the CWQB group (P ＜ 0.05).In the TMZ group and the CWQB group,early apoptosis rate was decreased.The decreasing in the TH group was more significant (P ＜ 0.05).The correlation analysis showed that the level of CK-MB and concentration of ADP had significant negative correlation.The early apoptosis rate and AMP had significant positive correlation.The early apoptosis rate had negative correlation with concentrations of ADP and ATP (P ＜ 0.01).It was concluded that CWQB recipe can decrease the concentration of AMP in H/R cardiomyocytes,increase the concentrations of ADP,ATP,TAN and EC,and decrease myocardial damage makers such as CK-MB and cTnT,depress early apoptosis rate in H/R cardiomyocytes,in order to display its cardioprotective effects in H/R injury.
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Objective To investigate the effect of procyanidins (PC) on cell apoptosis and cytochrome C releasing in human trabecular meshwork cells (HTMC) under oxidative stress induced by H2 O2.Methods HTMC were cultured and then divided randomly into 5 groups.Normal cultured HTMC served as normal group without treatment,and normal cultured HTMC in control group were treated with 500 μrnol · L-1 H2O2 for 1 h,as well as normal cells in treatment groups were treated with 500 imol · L-1 H2 O2 for 1 h combined with different concentrations of PC (0.02 g · L-1,0.05 g · L-1,0.10 g · L-1).Then flow cytometry and Western blot were applied to detect the cell apoptosis rate and the release of cytochrome C (CytC) respectively.Results Compared with the normal group,the apoptotic rates of the control group and the PC treatment groups were significantly increased,and the difference was statistically significant (all P ＜ 0.01).Compared with the control group,the apoptotic rates of the 3 PC treatment groups were decreased,and the differences were statistically significant (all P ＜ 0.01).Moreover,the apoptotic rate in 0.02 g · L-1,0.05 g · L-1,0.10 g · L-1 PC treatment groups was decreased as PC concentration increased,and the differences were statistically significant (all P ＜ 0.01).Furthermore,compared with the normal group,the release of cytochrome C in the control group,0.02 g · L-1 and 0.05 g · L-1 PC treatment group enhanced,and the differences were statistically significant (all P ＜ 0.01),but there was no significant difference between 0.01 g · L-1 PC treatment group and the normal group (P ＞ 0.05).Compared with the control group,the release of cytochrome C in the 3 PC treatment group was attenuated with significant differences (all P ＜0.01).Meanwhile,the release of cytochrome C was decreased in the 3 PC treatment groups with a concentration-dependent manner,and the pairwise differences were statistically significant (all P ＜ 0.01).Conclusion Exogenous PC can decrease the apoptotic rate of HTMC under oxidative stress and reduce the release of cytochrome C,as well as it also has an antioxidant effect in a certain concentration-dependent manner.
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Objective To investigate the effect of procyanidins (PC) on cell apoptosis and cytochrome C releasing in human trabecular meshwork cells (HTMC) under oxidative stress induced by H2 O2.Methods HTMC were cultured and then divided randomly into 5 groups.Normal cultured HTMC served as normal group without treatment,and normal cultured HTMC in control group were treated with 500 μrnol · L-1 H2O2 for 1 h,as well as normal cells in treatment groups were treated with 500 imol · L-1 H2 O2 for 1 h combined with different concentrations of PC (0.02 g · L-1,0.05 g · L-1,0.10 g · L-1).Then flow cytometry and Western blot were applied to detect the cell apoptosis rate and the release of cytochrome C (CytC) respectively.Results Compared with the normal group,the apoptotic rates of the control group and the PC treatment groups were significantly increased,and the difference was statistically significant (all P ＜ 0.01).Compared with the control group,the apoptotic rates of the 3 PC treatment groups were decreased,and the differences were statistically significant (all P ＜ 0.01).Moreover,the apoptotic rate in 0.02 g · L-1,0.05 g · L-1,0.10 g · L-1 PC treatment groups was decreased as PC concentration increased,and the differences were statistically significant (all P ＜ 0.01).Furthermore,compared with the normal group,the release of cytochrome C in the control group,0.02 g · L-1 and 0.05 g · L-1 PC treatment group enhanced,and the differences were statistically significant (all P ＜ 0.01),but there was no significant difference between 0.01 g · L-1 PC treatment group and the normal group (P ＞ 0.05).Compared with the control group,the release of cytochrome C in the 3 PC treatment group was attenuated with significant differences (all P ＜0.01).Meanwhile,the release of cytochrome C was decreased in the 3 PC treatment groups with a concentration-dependent manner,and the pairwise differences were statistically significant (all P ＜ 0.01).Conclusion Exogenous PC can decrease the apoptotic rate of HTMC under oxidative stress and reduce the release of cytochrome C,as well as it also has an antioxidant effect in a certain concentration-dependent manner.
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Objective: To study the inhibitory effect of total alkaloids from lotus seed on human hepatoma HepG2 cells.Methods: The effect of total alkaloids from lotus seed on the growth of HepG2 cells was studied by CCK-8 kit.The apoptosis rate of HepG2 cells was detected by flow cytometry.Results: When the action time was the same, with the increase of drug concentration, the inhibitory rate of total alkaloids from lotus seed on HepG2 cells increased, in a dose-dependent manner.At 72 h, the half inhibitory concentration (IC50) of total alkaloids from lotus seed on HepG2 cells was 1.501 μg·ml-1.At the same concentration, the inhibitory rate of the total alkaloids from lotus seed on HepG2 cells increased with the extension of the action time.At 72 h, the inhibition rate of 10 μg·ml-1 total alkaloids from lotus seed reached 72%.After treated with the total alkaloids from lotus seed at different concentrations, the apoptosis rate of HepG2 cells significantly increased in a dose-dependent manner.Compared with the blank control group, the difference was statistically significant (P <0.05), and the apoptosis rate of HepG2 cells was 85.6% treated with 20 μg·ml-1 total alkaloids from lotus seed.Conclusion: The total alkaloids from lotus seed can induce cell apoptosis and inhibit the proliferation of human hepatoma HepG2 cells.
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Objective To explore the protective effect of Zhenbao pill on acute spinal cord injury of rat and its possible mechanisms.Methods 64 Wistar rats were randomly divided into a injury group and a drug group, with 32 rats in each group, A T10 acute incomplete spinal cord injury model was produced by using modified Allen technique in the two groups,Zhenbao pill mixed suspension of 0.6 g/kg was given by gavage method daily in the drug group 30 min after moding, the same amount of saline was given in the injury group. Behavior detection were conducted in the first day, third day, and the seventh day after modeling to each 8 rats of the two groups. Spinal cord specimens was got centered about the damage points after rats were executed. histopathological changes were observed under HE staining, the MDA content were determined by thiobarbituric acid spectrophotometer and the apoptotic cells were marked by TUNEL method.Results After 1, 3, 7 d, compared with the injury group, the MDA level in the rat spinal cord tissue in the drug group (15.12 ± 1.27 nmol/mgvs.19.71 ± 1.29 nmol/mg, 20.42 ± 1.33 nmol/mgvs.24.65 ±1.36 nmol/mg, 10.46±1.49 nmol/mg vs.15.46 ± 1.44 nmol/mg) decreased(P<0.05); the apoptosis rate of drug groups (19.61% ± 1.49%vs.26.61% ±1.52%, 21.49% ± 1.37% vs.32.37% ± 1.24%, 13.04% ± 1.30%vs.18.35% ± 1.27%)decreased (P<0.05); the BBB score of drug groups (6.52 ± 1.27vs. 2.63 ± 1.27), (12.68 ± 1.32 vs.6.17 ± 1.34), (15.47 ± 1.27 vs.11.57 ± 1.29) was higher than the injury group (P<0.05). Conclusion Zhenbao pill can significantly improve the histopathological examination,reduce the MDA content in the injured spinal cord specimen of the rat and the rate of neuronal apoptosis, which may promote recovery of neurologic function in the rat suffered from ASCI.
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Objective To observe the influence of Zhibai Qianqing(ZBQQ) Decoction combined with Levofloxacin on sperm quality of chronic prostatitis(CP) patients and explore its possible mechanism through sperm apoptosis rate.Methods Fifty eight cases were divided into 2 groups randomly.Twenty eight cases were served with ZBQQ decoction combined with Levofloxacin(observe group),30 cases were treated by Levofloxacin tablets(control group).The curative effect of the two groups was compared one month later.All the cases underwent semen analysis.Sperm apoptosis rates were measured by TUNEL before and after the treatment.Results Sperm liquefaction time of CP group was significantly longer than the normal group(P0.05).The clinic cure rate of observe group was 35.71%,and overall effective rate was 85.71%,significantly higher than that of control group(P