الملخص
Objective To observe the role of tumor necrosis factor-α (TNF-α) and platelet-derived growth factor-B (PDGF-B) in kiwi fruit essence-mediated protection of radiation-induced lung injury (RILI) in rats. Methods 96 male healthy Sprague-Dawley rats were divided into normal control group, model group, and kiwi fruit essence treatment group(60 and 240 mg/kg) by the random number table method, with 24 animals in each group. The whole lungs underwent 6 MV X-ray irradiation (18 Gy) to induce RILI animal models in rats of the latter three groups. On the next day after irradiation, rats in the latter two groups were intragastrically administrated with 60 or 240 mg/kg kiwi fruit essence, once a day. The rats in the normal control and model groups were treated with 9 g/L sodium chloride solution. Eight rats in the latter three groups were randomly sacrificed on days 14, 28, and 56, while normal control rats were sacrificed on day 56 as the overall control. Blood samples were collected and separated. Serum concentrations of TNF-α and PDGF-B were detected using ELISA. The lung tissues were isolated for HE and Masson staining to evaluate alveolitis and pulmonary fibrosis (PF). The hydroxyproline (HYP) content in lung tissues was detected. The mRNA and protein expression of pulmonary TNF-α and PDGF-B were determined by quantitative real-time PCR and immunohistochemistry. Results Compared with the model group, treatment with 60 and 240 mg/kg kiwi fruit essence group significantly reduced alveolitis on days 14 and 28 as well as PF lesions on days 28 and 56. Compared with the normal control group, HYP content in the lung tissue of the model group increased on day 28 and day 56, while TNF-α and PDGF-B levels in the serum and lung tissues increased at each time point. Compared with the model group during the same period, 60 and 240 mg/kg kiwi fruit essence element treatment group reported the diminished levels of serum and pulmonary TNF-α on day 14 and day 28. Consistently, the lung tissue HYP content and serum and pulmonary PDGF-B levels on day 28 and day 56 were reduced. In addition, the above indicators in the 240 mg/kg kiwi fruit essence treatment group were lower than those for the 60 mg/kg kiwi fruit essence treatment group. Conclusion Kiwi fruit essence can alleviate RILI in rats, which is related to the down-regulation of TNF-α expression at the early stage and decreased PDGF-B level at the middle and late stages.
الموضوعات
Animals , Male , Rats , Fruit/metabolism , Lung/radiation effects , Lung Injury/prevention & control , Oils, Volatile , Proto-Oncogene Proteins c-sis/metabolism , Pulmonary Fibrosis , Rats, Sprague-Dawley , Tumor Necrosis Factor-alpha/metabolism , Actinidia/chemistryالملخص
AIM: To observe the effects of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), myeloperoxidase (MPO) and transforming growth factor-β1 (TGF-β1) of radiation-induced lung injury rats by Kiwi fruit essence (unsaturated fatty acid of actinidia chinesis planch seed oil). METHODS: According to random number table, 90 Sprague-Dawley rats were divided into the control group, model group, 60 mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil intervention group, 120 mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil intervention group, 240 mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil intervention group, 18 animals were included in each group. Except for the control group, rats in the remaining groups were constructed by 6MV-X-ray 18Gy full chest radiation, one week before modeling, the rats in the last 3 groups were given (60, 120, 240) mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil. The first two groups were given 0.9% sodium chloride solution by gavage, once a day in each rat. 14 days, 28 days, and 56 days after radiation, the rats were randomly sacrificed and their chests were cut, and ave lung tissue was saved. HE and Masson staining were performed to observe the pathological changes, and the content of SOD, GSH-Px, MPO was determined. The expression of TGF-β1 was analyzed by immunohistochemical staining. RESULTS: Compared with the model group, (60, 120, 240) mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil significantly reduced the degree of lung alveolitis and radiation pulmonary fibrosis, reduced the content of hydroxyproline (HYP), MPO, increased the antioxidant enzymes SOD and GSH-Px content, down-regulated the expression of TGF-β1.There were significant differences in the above-mentioned indicators among the intervention groups of (60, 120, 240) mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil group, and it was positively correlated with dosage. CONCLUSION: Unsaturated fatty acid of actinidia chinesis planch seed oil has a preventive effect on radiation-induced lung injury, and its mechanism may be related to the reduction of oxidative stress damage and down-regulation of TGF-β1 expression.
الملخص
Objective To study the effect of soluble, refolded, recombinant extracellular domain of the human Fc gamma receptor Ⅱ a (huFcγRⅡa) on the binding of human IgG to cells. Methods Extra-cellular domain of the huFcγRⅡ a gene was amplified from recombinant plasmid pe3huR Ⅱ by PCR and then cloned into pET-28a vector. The recombinant plasmid pETshuR Ⅱ was transformed into E. coli BL21 (DE3) after identified by PCR and doubly digested. The inclusion bodies of fusion protein were extracted and purified by washing, dissolved in 6 mol/L guanidine buffer, and refolded by rapid dilution technique. The refolding protein activity was tested by ELISA and flow cytometry. Results Extraceilular domain of the huFcγRⅡa gene was successfully cloned into pET-28a. The results of SDS-PAGE showed that the molecular mass (Mr) of the expressed protein was 24.8 × 103, and the expression rate was 30%. The purity of recom-binant shuR Ⅱ was up to 90% after washing. ELISA showed that the recombinant shuR Ⅱ was able to bind human IgG in a dose dependent manner, shuRⅡ could competitively inhibit the binding of human IgG to huFcγRⅡa expressed on the surface of COS-7 cells by flow cytometry. Conclusion The results demon-strate that it is possible to obtain large quantities of recombinant shuR Ⅱ which has comparable binding prep-erties to those of the whole membrane bound huFcγR Ⅱ a.