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Sheng Mai injection on paraquat poisoning in rat lung tissue nuclear factor-кB and inducible nitric oxide synthase expression in

Abstract Objective To observe the paraquat (PQ) induced lung injury in rats with acute poisoning (ALI), nuclear factor-кB and inducible nitric oxide synthase (iNOS) in the change of acute paraquat poisoning Shengmai induced lung injury. Methods 50 SD rats were randomly divided into 5 groups, control group, negative control, positive control group,[link widoczny dla zalogowanych], low dose group and Shengmai Shengmai high dose group. Observation of gross pathological specimens, simultaneous determination of pulmonary nuclear factor-кB and iNOS activity. Results Compared with the negative control group, low dose group Shengmai lung pathology showed lung congestion, significantly reduced pulmonary edema; nuclear factor-кB and iNOS also reduced (P
Key words Shengmai injection of paraquat lung injury of nuclear factor-κB nitric oxide nitric oxide synthase
Abstract: ObjectiveTo invesigate the protective effect of pulse-activating injection against rat acute lung injury.Methods 50 Sprague-Dawley (SD) rats were randomly divided into five experimental groups: blank group,[link widoczny dla zalogowanych], negative control group, positive control group, Pulse -activating injection low dose group (LDG), Pulse-activating injection high dose group (HDG). Macroscopic and histopathological examinations were performed, and the activities of lung tissue homogenate NK-кB and iNOS were measured. ResultsCompared with negative control group,[link widoczny dla zalogowanych], and biological indexes of lung injury and the contents of NK-кB and iNOS were significantly decreased in pulse-activating injection groups (P <0.05, P <0.01). ConclusionNK-кB and iNOS play important roles in lung injury of poisoned rats. Pulse- activating injection can ameliorate the lung injury and decrease the levels of NK-кB and iNOS.
Key words: Pulse-activating injection; Paraquat; Lung injury; NF-κB; Nitric oxide (NO); Inducible nitric oxide synthase (iNOS)
Paraquat (paraquat, PQ) weed its good effects are widely used worldwide, but the reach of or subsequent suicide brought the high mortality rate of oral health care gave workers made a tremendous challenge. Paraquat poisoning lethal dose is small, the maximum drug concentration in lung tissue, so the most severe lung injury, most patients died of respiratory distress syndrome. Treatment of a variety of ways people try to paraquat poisoning, but so far there is no effective antidote. The results show that paraquat causes lung injury led to one of the nuclear factor-кB and increased iNOS activity [1], but significantly lower Shengmai acute lung injury in nitric oxide levels and inducible nitric oxide synthase ( iNOS) activity, so students Injection for chronic obstructive pulmonary disease (COPD), chronic bronchitis and tuberculosis and the treatment of toxic shock, etc. [2,3], but for the treatment of paraquat poisoning has not been reported. Therefore, this experiment replicated Paraquat poisoning by rat model of Sheng Mai injection on acute lung injury induced by paraquat poisoning effect and mechanism of paraquat in clinical emergency provision of new relief drugs.
1 Material and Methods
1.1 Materials 20% PQ Concentrate (UK Department of Zeneca Limited pesticides); Health Injection (Set An Yisheng Pharmaceutical Co., Ltd. of Jilin Province, batch number: 080115); dexamethasone sodium phosphate injection (Zhengzhou Zhuo Feng Pharmaceutical). Monoclonal mouse anti-human NF-κB p65 antibody (SC-8008), SP immunohistochemistry kit (SP-9002) and Concentrated DAB kit (ZLI-9032) were purchased from Beijing Zhongshan Biotechnology Co., Ltd.; Nitric oxide kit,[link widoczny dla zalogowanych], nitric oxide synthase assay kit, Coomassie brilliant blue protein assay kit (built by the Nanjing Institute of Biotechnology). SHA-BA-type dual-function thermostatic water bath oscillator (Jintan Jeryl Electric Co., Ltd.); XW-80A-type vortex mixer (Shanghai Huxi Analytical Instrument Factory); UNIVERSAL-16R-based desktop centrifuge at low temperature ( Germany Hettich ENTRIFVGENTD); Stat Fax-2100 microplate type (AWARENESS TECHN-OLOGYING); CKX41 Inverted biological microscope (Japanese OLY-MPUS).
1.2 SD rats weighing animals and groups after the control group injected with normal saline 30 mg / kg, 20% of the other group were injected paraquat 30 mg / kg paraquat poisoning model of replication. Animal groups as follows: ① the control group (NS): normal saline 4 mg / kg; ② negative control group (PQ): intraperitoneal injection 4 mg / kg; ③ positive control group (D): intraperitoneal injection in Cyprus Betamethasone 0.5 mg / kg; ④ Shengmai low dose group (LDG): intraperitoneal injection Shengmai injection 0.2 ml / kg; ⑤ Shengmai high dose group (HDG): intraperitoneal injection Shengmai injection 0.4 ml / kg. More than 5 animals were administered 2 times / d, for consecutive 3 d.
1.3 water sample collection and processing co-chloralose anesthetized rats exposed trachea and lungs after the chest, abdominal aortic blood were killed, sacrificing serum; and immediately remove the lung reserve.
1.4 NO determination precision weigh a certain amount of lung tissue, a continuous 95% (volume fraction) O2-5% (volume fraction) CO2, 37 ℃ constant temperature incubation of 2 h, the supernatant obtained 100 μl, operating strictly according to kit instructions.
1.5 Determination of lung tissue iNOS activity in lung tissue proper speed, according to 1:5 (W / V) by adding extract of pre-cooling, ice bath, homogenized, 4 ℃, 3 000 r / min centrifugation 10 min, the supernatant were collected, with Coomassie brilliant blue kit protein, iNOS extraction kit iNOS activity measured.
1.6 NF-κB p65 in lung tissue was determined by immunohistochemical streptavidin avidin - biotin (LSAB) method staining NF-κB p65 in lung tissue and distribution situation. Selection of a rabbit anti-mouse anti-NF-κB p65 polyclonal antibodies, dilution 1:100,[link widoczny dla zalogowanych], with SP kit, followed by instructions to operate. 5 randomly selected microscopic high power field, calculated in 5 high power field positive cells (in membrane / cytoplasm / nucleus appears brown colored particles as the positive mark) percentage of total cells.
1.7 Histological examination taken a few of lung tissue in 10% formalin fixed, paraffin embedded sections, HE staining observed with a microscope of lung tissue injury.
1.8 Statistical analysis of experimental data with ± s said. Two or more data analysis using One-way ANOVA, group Newman-Student-Keuls test for statistical analysis; two sets of data processing using Student's t test. P <0.05 considered significant difference.
2 Results
2.1 lung tissue changes in NO and iNOS results show that, PQ lung tissue NO concentration and iNOS activity was significantly higher than the NS group (P <0.01), paraquat can cause lung tissue shows generate a lot of NO, and to increased iNOS activity in lung tissue. Application Shengmai, dexamethasone treatment intervention, the lung tissue NO concentration and iNOS activity significantly lower than the PQ group (P <0.01, P <0.05), so Shengmai inhibit NO production in lung tissue and inhibit iNOS activity, and Health effects of low dose pulse better. The results shown in Table 1 and Figure 1 and 2. Table 1 in the lung tissue NO, iNOS content changes (slightly)
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