Lung Bcl-2 Research Articles

Changes of autophagy in rat lung tissue after acute spinal cord injury

Objective To investigate the changes of autophagy in rat lung tissues after acute spinal cord injury (ASCI) and the possible mechanisms. Methods Sixty-three female SD rats were divided into sham operation group (n=21) and spinal cord injury group (n=42), according to the random number table. The modified Allen method with the impact energy of 10 × 25 g·mm at the T10 vertebra was used for preparation of ASCI models. The rats were sacrificed at 6, 12, 24, 48, 72 hours, 1 and 2 weeks after injury. Lung tissue damage and apoptosis were detected by HE staining and TUNEL method. The changes of autophagy and expressions of LC3-Ⅱ, P62, Beclin-1, interleukin (IL)-17A and Bcl-2 in lung were detected by Western blot and immunofluorescence. Results The pulmonary alveoli maintained normal structure in sham operation group, with no inflammation or pulmonary hemorrhage. Slight lung tissue damages were observed in spinal cord injury group at 12 h postinjury. Alveolar stroma widening, inflammatory infiltration, hemorrhage, and alveolar collapse became ingravescent at 24-72 hours postinjury. Numbers of apoptotic cells in spinal cord injury group were 551.22±135.94, 905.11±92.64, and 141.78±30.86 respectively at 24, 72 hours and 1 week postinjury, and were significantly increased at 24 and 72 hours postinjury, compared with sham operation group (P<0.05). Expression of LC3-II in spinal cord injury group was increased at 24-72 hours postinjury, compared with sham operation group (P<0.05). Expression of P62 in spinal cord injury group was up regulated at 24-72 hours postinjury, compared with sham operation group (P<0.05). Expression of Beclin-1 in spinal cord injury group was increased at 24 h postinjury and then dropped at 48-72 hours, compared with sham operation group (P<0.05). Expression of IL-17A in spinal cord injury group was increased at 24-48 hours, compared with sham operation group (P<0.05). Expression of Bcl-2 in spinal cord injury group was increased from 24 hours to 72 hours, compared with sham operation group (P<0.05). Conclusion Autophagosome formation is increased and accumulated in the lung tissues after ASCI, which might be related to the increased interaction between Beclin-1 and Bcl-2 because of the up regulation of Bcl-2 by IL 17A, ultimately leading to the inhibition of autophagy. Key words: Spinal cord injury; Lung; Autophagy; Interleukin -17A

New Contributions to Asarum Powder on Immunology Related Toxicity Effects in Lung.

Objective. Asarum is widely used in clinical practice of Chinese medicine in the treatment of respiratory diseases. Many toxic ingredients (safrole, etc.) had been found in Asarum that show multiple visceral toxicities. In this study, we performed systematic investigation of expression profiles of genes to take a new insight into unclear mechanism of Asarum toxicities in lung. Methods. mRNAs were extracted from lungs of rats after intragastric administration with/without Asarum powders, and microarray assays were applied to investigate gene expression profiles. Differentially expressed genes with significance were selected to carry out GO analysis. Subsequently, quantitative PCRs were performed to verify the differential expression of Tmprss6, Prkag3, Nptx2, Antxr11, Klk11, Rag2, Olr77, Cd7, Il20, LOC69, C6, Ccl20, LOC68, and Cd163 in lung. Changes of Ampk, Bcl2, Caspase 3, Il1, Il20, Matriptase2, Nfκb, Nptx2, and Rag2 in the lung on protein level were verified by western blotting and immunohistochemistry. Results. Compared with control group, the estimated organ coefficients were relatively increased in Asarum group. Results of GO analysis showed that a group of immune related genes in lung were expressed abnormally. The result of PCRs showed that Ccl20 was downregulated rather than other upregulated genes in the Asarum group. Western blotting and immunohistochemistry images showed that Asarum can upregulate the expression of Ampk, Caspase 3, Il1, Il20, Matriptase2, Nfκb, and Rag2 and downregulate the expression of Bcl2 in lung. Conclusion. Our data suggest that expressions of immune related genes in lung were selectively altered by Asarum. Therefore, inflammatory response was active, by regulating Caspase 3, Il1, Il20, Matriptase2, Nfκb, Rag2, Tmprss6, Prkag3, Nptx2, Antxr1, Klk11, Olr77, Cd7, LOC69, C6, LOC68, Cd163, Ampk, Bcl2, and Ccl20. Our study indicated that inflammatory factors take effect in lung toxicity caused by Asarum, which provides a new insight into molecular mechanism of Asarum toxicities in lung.

P53, Bcl-2 and CD68 expression in response to amethopterin-induced lung injury and ameliorating role of l-carnitine

Amethopterin (methotrexate, MTX) is an antimetabolite and antifolate drug with antiflammatory properities and is used to treat autoimmune diseases, such as psoriasis, rheumatoid arthritis and certain types of cancer, such as breast, lymphoma and lung. The present study aimed to study the changes in P53, Bcl-2 and CD68 expression in response to amethopterin-induced lung injury and ameliorating the role of l-carnitine. A total of 36 male albino rats were equally divided into six groups: the first and second groups were the control and l-carnitine groups respectively while the 3rd group was amethopterin rat group; the 4th and 5th groups were co- and post-treated amethopterin rat with l-carnitine respectively and the 6th group was self treated amethopterin rat group. Our results shows that lung in amethopterin-treated rats showed many of histopathological alterations as severe to strong alveolar damage in the form of collapsed alveoli and strong thickened interalveolar septa with heavy infiltration of inflammatory cells. This damage was increased or remaining in self-amethopterin-treated group. Treatment (co- and post) with l-carnitine were improved in the lung structure that was treated with amethopterin. A significant increase in p53 and CD68 and decrease in Bc1-2 immunoreactivity in the lung in amethopterin group is observed when compared with the control group. However, treatment of rats with l-carnitine decreased the intensity of P53-ir and CD68-ir and increased the intensity of Bcl-2 in lung when compared with amethopterin rat group. Co-treatment with l-carnitine improved lung damage induced with amethopterin.

Evaluation of nestin in lung adenocarcinoma: relation to VEGF and Bcl-2

Context: Nestin is a marker of multipotent precursor cells that is up regulated in cancer.Objective: To explore its diagnostic role and its relationship to vascular endothelial growth factor (VEGF) and Bcl-2 in lung adenocarcinoma.Materials and methods: Evaluation of nestin expression in lung biopsies by real-time PCR and serum VEGF and Bcl-2 by ELISA in 27 adenocarcinoma patients and 15 control subjects.Results: Nestin was significantly higher in lung adenocarcinoma patients especially with advanced grade and stage and it was significantly correlated to VEGF and Bcl-2.Conclusion: Nestin can be considered as a potential diagnostic marker in lung adenocarcinoma.

Effects of sulfur dioxide on expressions of p53, bax and bcl-2 in lungs of asthmatic rats

Inhibition of cell apoptosis is an increasingly important factor in modulating airway inflammation in asthma, which is related to environmental pollutants. To investigate the effects of sulfur dioxide (SO2) on the mRNA and protein expressions of apoptosis-related genes in lungs from asthmatic rats, male Wistar rats were challenged by ovalbumin (OVA) or SO2 (2 ppm) inhalation alone or together. Examinations were performed 24 h after the last treatment. The mRNA and protein levels of p53, bax, and bcl-2 were analyzed in lungs using real-time reverse transcription-polymerase chain reaction (RT-PCR) assay and Western blot analysis, respectively. The results indicated that increases of bcl-2 or decreases of p53 and bax mRNA and protein levels were not significant in lungs of rats exposed to SO2 alone, compared with controls, but elevated or reduced levels of these genes appeared in lungs of asthmatic rats exposed to SO2 plus OVA, compared with controls, suggesting that SO2 exposure could result in OVA-induced increases or decreases of transcription and translation levels of these apoptosis-related genes in rat lungs, and may have relations to airway inflammation in asthma. The regulation mechanism of apoptosis in asthma disease exposure to SO2 needs further study.

Immunomodulatory and antitumour effects of an exopolysaccharide fraction from cultivated Cordyceps sinensis (Chinese caterpillar fungus) on tumour‐bearing mice

Cordyceps sinensis (Chinese caterpillar fungus) is a fungus parasitic on the larvae of Lepidoptera and has been considered to be a precious tonic food and herbal medicine since ancient times in China. Recently, some fungal strains have been isolated from the fruiting bodies of wild C. sinensis, and some of them have been reported to show the same properties as the natural product. In the present study, an EPSF (exopolysaccharide fraction) was prepared from cultivated C. sinensis and its effects on B16 melanoma-bearing mice were investigated. Three doses of EPSF were intraperitoneally administered every 2 days after the day of tumour-cell injection. The experiment was terminated on day 28. Phagocytosis of peritoneal macrophages and proliferation of spleen and thymus lymphocytes were assayed. The tumour metastatic foci on the lung and liver surface were checked. The expression of oncoprotein Bcl-2 in livers and lungs was assayed by a immunohistochemical method. The results showed that EPSF significantly enhanced the Neutral Red uptake capacity of peritoneal macrophages (60 mg/kg, P<0.01; 120 mg/kg, P<0.001) and spleen lymphocyte proliferation (60 mg/kg, P<0.05; 120 mg/kg, P<0.001) in B16-bearing mouse. The metastasis of B16 melanoma cells to lungs (120 mg/kg) and livers (30, 60 and 120 mg/kg) was significantly inhibited by EPSF. Moreover, EPSF decreased the levels of Bcl-2 in the lungs (120 mg/kg) and livers (30, 60 and 120 mg/kg). These results suggest that EPSF has immunomodulatory function and antitumour activity.

Induction of apoptosis in the lung but not in the liver of rats receiving intra-tracheal instillations of chromium(VI).

Several studies have shown that hexavalent chromium [Cr(VI)] induces apoptosis in a variety of in vitro test systems. We instilled intra-tracheally either saline or sodium dichromate (0.25 mg/kg body weight), for three consecutive days, to Sprague-Dawley rats. TUNEL analyses showed a marked increase of the apoptotic index in both bronchial epithelium and lung parenchyma of Cr(VI)-treated rats, but no effect was detected in their liver. In parallel, the expression of 13 out of 18 apoptosis-related genes, evaluated by cDNA array analysis, was significantly enhanced in rat lung. The overexpressed genes included c-Jun N-terminal kinases 1, 2 and 3, bcl-x, bcl-2-associated death promoter and bcl-2-related ovarian killer protein, caspases 1, 3 and 6, DNase I precursor, DNA topoisomerases I and II alpha, and poly(ADP-ribose) polymerase. The enhancement of p53 expression in the lung was borderline to statistical significance. Expressions of bcl-2, bax-alpha, mdm2 and DNA topoisomerase IIB were not enhanced to a significant extent in lung. No induction of gene expression was observed in rat liver. RT-PCR analyses confirmed that Cr(VI) enhances the expression of c-Jun N-terminal kinase 1, caspase 6, and DNase I precursor but not that of bcl-2 in lung, while none of these genes was overexpressed in the liver of Cr(VI)-treated rats. The lack of stimulation of apoptosis in the liver parallels the failure of Cr(VI) to produce genotoxic damage, as we previously observed under identical experimental conditions. These negative findings may be ascribed to reduction of Cr(VI) to Cr(III) when traveling from the respiratory tract to the liver. On the other hand, induction of apoptosis in the respiratory tract parallels the occurrence of genotoxic effects and oxidative DNA damage produced by Cr(VI) in the same tissue. As previously shown in another laboratory, Cr(VI) did not induce lung tumors after 30 months of administration of the same daily dose. Therefore, apoptosis is likely to provide a protective mechanism at a post-genotoxic stage of Cr(VI) carcinogenesis.