Artesunate On Paraquat-induced Pulmonary Injury Mechanism Of Intervention Studies

Objective:In recent years, paraquat poisoning reports are presenting an increasing trend ,which owns the feature of toxic dose small, nor-effective antidote ,nor-effective therapy, which leads to the increased mortality of PQP. After poisoning, it mainly prominently damange the lungs of patients.But the exact injury mechanism is currently not clarified.Via inspecting the level of tumor necrosis factorα(TNF-α) and transforming growth factorβ₁ (TGF-β₁) of Serum cytokines and histopathological changes of lungs of Paraquat poisoning mouse under the Intervention of traditional Chinese medicine– artesunate, this experiment is aimed to research the possibility and evolution of lungs damages of paraquat poisoning mouse and to study the Intervention role of artesunate to the lungs damages of lungs of paraquat poisoning mouse in order to discover how the paraquat harms the lungs of mice and to find an ideal target for treatment and drugs to make a useful attempt.Method:A total of 65 SD male mice were randomly divided into two groups,20 in control group A and 45 in poisoning group B. Group B was divided into two groups: artesunate group(22mice) and non-artesunate group (23 mice). After fasting for 12 hours, the45 mice in group B were fed by gastric gavage[1] with 50mg/kg paraquat which was diluted to 1ml, the 20 mice in group A were given 1ml normal saline by gastric gavage. Then,the mice in group Z were given intraperitoneal injection of 10 mg/100 g/d of artesunate solution. The mice in group A and group P were given intraperitoneal injection of the same volume of normal saline.Randomly Chose 5 mice in group A, 5 mice in group Z and 5 mice in group P since 12 hours, 24 hours, 48 hours and 72 hours after the experiment. Those chosen mice were laparotomized when anesthetized with ether inhalation. Extracted 5ml blood in the abdominal aorta, centrifuged the blood and kept the supernatant sepamouseely.Tested the concentmouseion of TNF-αand TGF-β₁ in mice Serum of the 3 selected group with Enzyme-linked immunosorbent assay method(ELISA) and got the OD result on 450nm. Used SPSS 17.0 statistical package to analyze the results. Made comparison between the two groups and the group mice at different time points of serum TNF-αand TGF-β₁ dynamic changes. All results were presented as mean±standard deviation,analysed of variance between groups. P <0.05 was considered statistically significant. Then anatomise the male mice who were drawn blood and,observed the changement of lungs.Then took a small part of the left lung tissue,soaked it in 10% formaldehyde solution,used paraffin-embedded, made pathological section and Hematoxylin– eosin staining and finally for light microscopic examination.Results:1.At different times, the poisoning mice showed listlessness, lack of exercise, irritability, anorexia, irritability, fluffy, and shortness of breath, which were similar to the clinical manifestations in previous experiments.[2] 2. After poisoning, the level of TNF-αand TGF-β₁ of Serum cytokines in those poinsoning mice increased gradually with prolonged poisoning dumouseion.①The comparison between non-artesunate group and control group:the level of TNF-αand TGF-β₁ of serum cytokines in non-artesunate group increased obviously since 12 hours after poisoning, then it increased gradually and reached the maximum in the 72nd hour. Those figures were significant different from the control group (P<0.01).②The comparison between artesunate group and control group:the level of TNF-αand TGF-β₁ of serum cytokines in artesunate group increased obviously since 12 hours after poisoning(P<0.01), then it increased gradually and reached the maximum in the 72nd hour. Those figures were significant different from the control group.③The comparison between artesunate group and non-artesunate group:the level of TNF-αand TGF-β₁ of serum cytokines in artesunate group was lower than non-artesunate group between 12 hours and 72 hours after poisoning. However, there was the differences at the 48th hour and 72nd hour had statistical significance ( P<0.05).3.①The appearance of lungs of mice: the color of lungs in normal mice were bright pink and the texture of lungs were soft; the lungs of non-artesunate group were dark red and obvious swelling with blunt edges and lung congestion;the lungs of artesunate group were also swelling with obvious lung congestion but the color was slightly shallower than non-artesunate group and the swelling and congestion were milder than non-artesunate group .②Observed the pathological section and Hematoxylin - eosin staining of lungs:the lung structure of control group was normal。The mice in poinsoning group presented expansion of the alveolar capillary and inflammatory cell infiltmouseion in 6 hours; showed expansion of the alveolar capillary and Infiltmouseion of neutrophils in the 24th hour;reached the summit of Pathological changes in the 48th hour;declined a little bit in the 72nd hour. The lungs of mice in artesunate group and non-artesunate group had similar pathological changes. But after 24 hours, the lungs injury of artesunate group was mitigated at the same time point. Anyway, the difference wasn't that obvious.Conclusions:①The level of both TGF-β₁ and TNF-αof Serum cytokines of lungs of Paraquat poisoning mice increased. This rise became obvious after 12 hours and reached the summit in the 72nd hour. This showed that the cytokine TGF-β₁ and TNF-αtook part in the occurrence and development of acute lung injury of mice by paraquat poisoning.②Took artesunate intervention treatment to reduce TGF-β₁ and TNF-αlevels and acute lung injury level of paraquat poisoning mice.