| Objective Establishing rat models of radiation induced lung injury,to explore the self-prepared prescription’s mechanism by observing its effect on general condition, pathological changes of lung tissue, TLR4 signaling pathway proteins and related cytokines of rats caused by radiation.Methods The self-prepared anti-pulmonary fibrosis prescription is composed of radix astragali, radix adenophorae, coastal glehnia root, lily, caulis dendrobii, phlegm, loquat leaf, almonds, radix stemonae, angelica,which appear to be different quality proportion of three different doses recipes, including TCM1,2 (the most common clinical dose) and 3.84 SD male rats were randomly divided into irradiation+distilled water group, irradiation+3 TCM groups, irradiation+dexamethasone group, normal group and normal TCM2 group, a total of seven groups. They were put into experiment after adaptive feed 7 days, then fasting but water for 24h.When finished irradiation,those rats were fed with TCM, dexamethasone or distilled water by 1ml/100g and noticed their mental states, appetites, color, weights and other general situation changes.After continuous feeding 14 days, putting every rat to death,taking out and macroscopicly observing lung tissue,then contrasting pathological changes after HE staining microscopic observation and scoring injury degree.Choped and gringed in aseptic conditions, lung tissue liquid supernatant was detected the expression of protein molecules MyD88, TRAF-6 andTRIF, TLR4 of TLR4 signaling pathway on the western blot and biological image gray-scale analysis test and TNF-α, IL-17 and TGF-beta on the ELISA method. SPSS17.0 statistical software was used for data analysis.Results 1. General condition of rats 90% irradiation group rats appeared depression, loss of appetite, lackluster hair, yellow, dry, erectand even hair removal and other symptoms. The general states of rats treated with TCM2,3 and dexamethasone were better than those of rats cured with TCM1 and distilled water.2. The body weight of rats After treatment with TCM2,3 and dexamethasone, irradiation group rats’ weight increased significantly(p<0.05) compared with TCM1 and distilled water group rats’,while there was no statistical significance(p>0.05) between TCM2,3 irradiation group rats ’weight and dexamethasone irradiation group rats’.3. Pathological changes The irradiation group rats present to be acute inflammation, The lesions are mainly composed of seepage.Major pathological manifestations were:lung thick septa, the alveolar walls broadening, alveolar wall capillary hyperemia, pulmonary interstitial hyperemia, edema, alveolar interval and part of the alveolar lumen infiltrating scattered neutrophils. Visually, the inflammation response of TCM2,3 and dexamethasone irradiation group rats reacted gently compared with TCM1 and distilled water group rats.4. Pathological scores Pathologic grades showed irradiation group rats was obviously higher than that of normal group rats, with statistically significant difference (p< 0.01). What’s more, the scores of TCM2,3 and dexamethasone irradiation group rats were less than those of TCM1 and distilled water group rats.While there was no statistical significance(p>0.05) between TCM2,3 irradiation group rats and dexamethasone irradiation group rats.5. Activity of TLR4 signaling pathway With the methods of western blot and biological image gray level analysis,compared with normal group rats,the expression of MyD88,TRAF-6 and TRIF,TLR4 of irradiation group rats increased obviously.Moreover,the amounts of MyD88, TRAF-6 and TRIF, TLR4 of TCM2,3 and dexamethasone irradiation group rats were less than those of TCM1 and distilled water group rats.6. Expression of some cytokines Contrasted with normal group rats by elisa testing, the expression of TNF-a, IL-17, TGF-β of irradiation group rats increased obviously more than that of normal group rats, with statistically significant difference (p< 0.05). Furthermore, the quantities of TNF-a, IL-17, TGF-β of TCM2,3 and dexamethasone irradiation group rats were lower than those of TCM1 and distilled water group rats.While there was no statistical significance(p>0.05) between TCM2,3 irradiation group rats and dexamethasone irradiation group rats.Conclusion Self-prepared prescription can improve the quality of life of rats, significantly reduce the reaction caused by radioactive pneumonia, relieve pulmonary interstitial hyperemia, exudation, hemorrhage, edema and alveolar structure damage.It may prevent or delay the development of the radiation pneumonia by inhibiting TLR4 signaling pathways and reducing cytokine expression in lung local tissue. In addition, the short-term effect is almost equal to hormones, and TCM2 concentration is considered to be the optimal concentration. |
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