Suppression of lung inflammation by the ethanol extract of Chung-Sang and the possible role of Nrf2.

BACKGROUND: Asian traditional herbal remedies are typically a concoction of a major and several complementary herbs. While balancing out any adverse effect of the major herb, the complementary herbs could dilute the efficacy of the major herb, resulting in a suboptimal therapeutic effect of an herbal remedy. Here, we formulated Chung-Sang (CS) by collating five major herbs, which are used against inflammatory diseases, and tested whether an experimental formula composed of only major herbs is effective in suppressing inflammation without significant side effects. METHODS: The 50% ethanol extract of CS (eCS) was fingerprinted by HPLC. Cytotoxicity to RAW 264.7 cells was determined by an MTT assay and a flow cytometer. Nuclear NF-κB and Nrf2 were analyzed by western blot. Ubiquitinated Nrf2 was similarly analyzed following immunoprecipitation of Nrf2. Acute lung inflammation and sepsis were induced in C57BL/6 mice. The effects of eCS on lung disease were measured by HE staining of lung sections, a differential cell counting of bronchoalveolar lavage fluid, a myeloperoxidase (MPO) assay, a real-time qPCR, and Kaplan-Meier survival of mice. RESULTS: eCS neither elicited cytotoxicity nor reactive oxygen species. While not suppressing NF-κB, eCS activated Nrf2, reduced the ubiquitination of Nrf2, and consequently induced the expression of Nrf2-dependent genes. In an acute lung inflammation mouse model, an intratracheal (i.t.) eCS suppressed neutrophil infiltration, the expression of inflammatory cytokine genes, and MPO activity. In a sepsis mouse model, a single i.t. eCS was sufficient to significantly decrease mouse mortality. CONCLUSIONS: eCS could suppress severe lung inflammation in mice. This effect seemed to associate with eCS activating Nrf2. Our findings suggest that herbal remedies consisting of only major herbs are worth considering.

Expression analysis of the pistil genes in controlling self-incompatibility of brassica campestris by CO2 gas using microarray.

In Brassica campestris, self-incompatibility (SI) can be overcome by CO2 gas treatment. Previously, we reported inhibition of the SI in the Brassica campestris line, 734, by CO2 gas. In this study, we used a cDNA microarray comprising 1,184 unique cDNA ESTs from a pistil-specific cDNA library of the Brassica line to examine the expression of the pistil genes in response to CO2. Microarray analysis revealed that expression of 2% of the pistil genes was altered by exposure to CO2 gas. Surprisingly, the SLG gene of the susceptible line was one of the down-regulated genes. Transmission electron microscopic analysis showed that the CO2 caused morphological changes in the papillary cell. Our results demonstrate that the overcoming of SI by CO2 gas involves reduced expression of the SLG gene in Brassica.

Identification of anther-specific gene expression from T-DNA tagging rice.

We have screened a total of 5,500 T-DNA tagging rice lines in which beta-glucuronidase (GUS) gene sequence was randomly inserted as a transgene into the plant genome. Histochemical GUS assays were carried out to select the T-DNA tagging rice lines that show its expression in anther. Of the tagging lines screened, three lines were found to express GUS specifically in the anther that is about 0.05%. Microscopic observation of the anther-expressed lines showed specific expression patterns of GUS in the anther, either gametophytic or sporophytic specificities. Southern blot analysis revealed that the integration copy number of the transgene was 2.3 in average. The detailed expression patterns were analyzed and discussed.