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Objective@#To explore the protective effect and mechanism of Somatostatin (SS) on the mice with Paraquat (PQ) poisoning, and to provide theoretical basis for clinical treatment of PQ poisoning.@*Methods@#48 SPF male BALB/c mice were randomly divided into control group, SS group, PQ group and PQ+SS group, with 12 mice in each group. 20 ml/kg SS solution was intraperitoneally injected into the SS group and PQ+SS group, and the same amount of normal saline was intraperitoneally injected into the PQ group and control group. After 1 hour of the above treatment, PQ group and PQ+SS group were given 60 mg/kg PQ solution by one-time gavage, while the control group and SS group were given the same amount of normal saline by gavage. After the above treatment for 3 hours, the SS group and PQ+SS group were intraperitoneally injected with SS solution (20 ml/kg) again, and the PQ group and the control group were intraperitoneally injected with the same amount of normal saline. 6 eyeballs were randomly selected from each group for blood collection, and the levels of TNF-α, MPO and il-6 in the blood of mice were detected by ELISA and other methods. The left lung was taken after blood collection to calculate the D/W ratio. The levels of SOD, caspase-3 and MDA were detected in some lung tissues by chemical colorimetry, and the amount of NF-κB was detected by Western blot. The lung histopathological changes were observed under light microscope.@*Results@#The mice in the control group and SS group showed normal activity and good general condition; Mice in the PQ group ate less and moved less, responded slowly to stimulation, breathed shallow and fast with thickened breath sound, had messy and dull fur, and had varying degrees of cyanosis on their lips and limbs; The above performance of PQ+SS group was less than that of PQ group. Under the light microscope, the alveolar structure of PQ group was disordered and seriously damaged. The pathological changes of lung tissue in PQ+SS group were significantly improved compared with that in PQ group, and the pathological scores were decreased (all P<0.05) . Compared with the control group, there was no significant change in the levels of various indicators in the SS group (all P>0.05) . While the levels of inflammatory cytokines (IL-6, TNF-α, Protein expression of NF-κB) , oxidative cytokines (MPO, MDA) and apoptotic cytokines caspase-3 in PQ mice were significantly increased, and the levels of oxidative cytokines SOD and lung tissue D/W were significantly decreased (all P<0.05) . Compared with the PQ group, the PQ+SS group, the levels of inflammatory cytokines (IL-6, TNF-α, Protein expression of NF-κB) , oxidative cytokines (MPO, MDA) and apoptotic cytokines caspase-3 decreased, and the levels of oxidative cytokines SOD and lung tissue D/W increased (all P<0.05) .@*Conclusion@#PQ poisoning can cause lung injury in mice, while SS can alleviate lung injury in PQ poisoned mice, improve the general survival state of mice, and play a certain protective role in lung injury caused by PQ poisoning, which may be achieved by SS throμgh reducing the inflammatory response, oxidative stress response and lung tissue apoptosis in lung injury caused by PQ poisoning.
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Objective To investigate the effect of Xuebijing on paraquat-induced oxidative stress and inflammation level in human kidney cell line-2 (HK-2) cells. Methods The HK-2 cells were routinely cultured and divided into blank control group, paraquat poisoning model group, Xuebijing injection pretreatment group and Xuebijing control group according to random number table method. The concentration of paraquat in HK-2 cells were measured by high performance liquid chromatography (HPLC); the activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) were measured by chemical colorimetry method; the levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA). Results With prolonged treatment, paraquat concentration gradually increased in HK-2 cells in the paraquat poisoning model group and the Xuebijing injection pretreatment group, reaching the peak value after treatment for 48 hours, moreover, the concentration of paraquat in HK-2 in the Xuebijing injection pretreatment group was significantly lower than that in the paraquat poisoning model group (mg/L: 4.26±0.20 vs. 5.77±0.18, P < 0.05). Compared with the blank control group, the contents of MDA, TNF-α and IL-6 of paraquat poisoning model group were obviously elevated [MDA (nmol/mg): 4.47±0.10 vs. 2.21±0.08, TNF-α (ng/L): 206.91±13.22 vs. 98.14±5.67, IL-6 (ng/L): 253.33±5.22 vs. 116.97±13.54, all P <0.05], and the activity of SOD was significantly reduced (U/mg: 33.30±0.62 vs. 41.58±0.17, P < 0.05). Compared with paraquat poisoning model group, the contents of MDA, TNF-α and IL-6 in Xuebijing injection pretreatment group were obviously decreased [MDA (nmol/mg): 2.92±0.17 vs. 4.47±0.10, TNF-α (ng/L): 166.29±15.47 vs. 206.91±13.22, IL-6 (ng/L): 209.39±3.18 vs. 253.33±5.22, all P < 0.05], and the activity of SOD was significantly increased (U/mg:38.10±0.67 vs. 33.30±0.62, P < 0.05) in the paraquat pretreatment group. Conclusion Xuebijing could reduce the concentration of paraquat in HK-2 cells, and decrease oxidative stress and inflammation factor levels in HK-2 cells.
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Objective@#To investigate the protective effect and mechanism of Xuebijing (XBJ) on paraquat (PQ) -induced apoptosis in Human kidney cell line-2 (HK-2) cells.@*Methods@#Routinely cultured HK-2 cells, (1) Cell growth inhibition experiment after PQ and XBJ intervention: PQ was divided into 0、200、400、800、1600 and 3200 μmol/L PQ groups, and the cell survival rate was detected after intervening 24、48 and 72 h. XBJ was divided into 0、5、10、20、40 mg/ml XBJ groups, and the cell survival rate was detected after intervening 24、48 and 72 h.To determine the rational drug concentration and the duration of action of XBJ and PQ. (2) PQ-induced HK-2 cell growth inhibition experiment antagonized by XBJ: The cells were divided into normal control group, PQ group (800 μmol/L) and PQ+XBJ group (The cells were pretreated with 5、10 and 20 mg/ml XBJ for 1 h, then cultured with PQ of 800 μmol/L) , After cultured 24 h、48 h and 72 h separately, the cell survival rate was detected. (3) HK-2 cells were divided into normal control group、PQ group (800 μmol/L PQ cultured for 24 h) 、PQ+XBJ group (pretreated with 10 mg/ml XBJ for 1 h, and then 800 μmol/L PQ cultured for 24 h) and XBJ group (10 mg/ml XBJ cultured 24 h). The apoptosis of cells was detected by flow cytometry. The protein expression of Bcl-2 and BAX in each group was detected by Western blotting. The expressions of caspase-3 and caspase-9 were detected by caspase-3 and caspase-9 activity kit active.@*Results@#(1) PQ could significantly reduced the survival rate of HK-2 cells and showed time and concentration dependence. The survival rate of HK-2 cells was about 55% after 800 μmol/L PQ contacted 24 h, XBJ under 20 mg/ml was no significant effect on the survival rate of HK-2 cells after cultured 72 h. (2) Compared with the PQ group, the survival rate of HK-2 cells of PQ+XBJ group was significantly increased (P<0.05). (3) Compared with the normal control group, the cell apoptosis rate of PQ group was significantly increased (P<0.05). Compared with the PQ group, the cell apoptosis rate of PQ+XBJ group was significantly decreased (P<0.05). (4) Compared with the normal control group, Bcl-2 protein expression in PQ group was significantly decreased and BAX protein expression in PQ group was significantly increased (P<0.05) ; compared with PQ group, Bcl-2 protein expression in PQ+XBJ group was significantly increased, BAX protein expression in PQ+XBJ group was significantly decreased (P<0.05). (5) Compared with the normal control group, the activities of caspase-3 and caspase-9 in PQ group were significantly increased (P<0.05). Compared with PQ group, the activities of caspase-3 and caspase-9 in PQ+XBJ group were decreased significantly (P<0.05) .@*Conclusion@#XBJ (10 mg/ml) has obvious protective effect on HK-2 cell injuried by PQ (800 μmol/L) , It can improve the survival rate of cells through reducing the apoptosis of HK-2 cells which induced by PQ.
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Objective@#To explore prognostic factors of acute paraquat poisoning (APP) , analyze the correlation between base excess (BE) and plasma concentration of paraquat (C-PQ) and discuss BE level in evalua-tion of prognosis of acute paraquat poisoning patients.@*Methods@#We retrospectively selected 84 APP patients who admitted to Emergency Intensive Care Unit (EICU) of our hospital from 2009.9.1 to 2015.8.31.Clinical data from 84 APP patients were analyzed. BE、C-PQ、time of gastric lavagesince ingestion、time of hemoperfusion since ingestion、severity index of paraquat poisoning (SIPP) 、white blood cell (WBC) 、percentage of neutrophils (N%) 、hemoglobin (HB) 、creatinine (Cr) 、alanine aminotransferase (ALT) 、aspartate aminotransferase (AST) 、partial pressure of oxygen (PaO2) 、partial pressure of carbon dioxide (PaCO2) and other laboratory parameters were measured. A total of 41 patients in non-survivors died during the 30 days after admission and 43 patients in survivors survived during the 30 days. The factors of prognosis in paraquat poisoining and the role of BE in evalu-ating prognosis was analyzed, as well as the correlation between BE and C-PQ.@*Results@#1.Logistic regression analyses showed BE、C-PQ、ALT、AST、Cr was of prognostic significance[odds ratio (OR) of BE: 0.511, 95%CI 0.267, 0.978; C-PQ:-=0.999, 95%CI 0.999, 1.000; both P<0.05] ; 2.The area under the receiver operating characteristic curve (ROC curve) of BE、C-PQ and prognosis were 0.775、0.927 respectively, BE≤-1.7 mmol/L was the best cut-off value, the sensitivity、specificity for predicting were 82.9%、62.8%, the evaluation value was lower to C-PQ>3 273.935 ng/ml (AUC 0.927, 78.0%、95.3%) ; 3.BE negative correlated with C-PQ[-1.100 (-4.100, -0.200) , -5.900(-8.650, -2.500) , both P<0.05]. (r=-0.4, P<0.01).@*Conclusion@#These results suggest that BE may be useful for the prediction of prognosis in PQ poisoning and BE negative correlated with C-PQ.