Research result of anti-inflammatory effects of a traditional medicinal plant in drosophila

In vivo screening for anti-inflammatory effects of traditional medicinal plant extracts. The intestinal epithelium is susceptible to damage caused by pathogens, oxidative stress, and toxic compounds. Foods containing SDS or NaCl could cause injury to the intestines and result in a melanotic phenotype in Drosophila. To screen for anti-inflammatory activities of traditional medicinal plant extracts, flies were fed a standard cornmeal medium supplemented with (experimental groups) or without (control group) aqueous extracts of the medicinal plants. Adult flies from each of the culture conditions were orally treated with the inflammatory reagent SDS or NaCl. First, a vial containing 30 adult flies from each culture condition was treated with 0.6% SDS containing 5% sucrose, and the survival rate was assessed over 6 days. The control group showed more than 88% mortality; however, some flies in the experimental groups appeared to have an increased survival rate. Out of 50 different medicinal plant extracts, 15 species significantly increased the survival rate by more than 50% compared with the control group. In addition, after treatment with 0.4 M NaCl, 18 species in which the survival rate increased by 50% over that of controls were also screened.

In other experiments, four plant extracts that showed a higher fly survival rate after treatment with SDS or NaCl (i.e., C. pilosula, S. lappa, I. cylindrical var. major, and M. toosendan) were selected for use as the test extracts. After treatment with SDS for 6 days, the survival rates of the experimental groups were 94.4%, 92.1%, 92.1%, and 76.6%, respectively, which were significantly higher than the survival rate of the control group (11.17%). Likewise, all experimental groups showed significantly increased survival rates (84.4%, 66.6%, 57.7%, and 65.5%) after treatment with 0.4 M NaCl. We then analyzed another inflammatory reagent, dextran sulfate sodium (DSS), which interferes with intestinal barrier function and stimulates local and systemic inflammation, causing similar tissue damage in adult Drosophila gut. Dramatically increased survival rates of 35.5%, 60%, 51.1%, and 61.1% were observed for extracts of these medicinal plants compared with the control group (1.1%). The results indicate that extracts of C. pilosula, S. lappa, I. cylindrical var. major, and M. toosendan are able to increase Drosophila survival rate after exposure to toxic compoundshttp://www.cusabio.com/.

Medicinal plant extracts can increase AMP leves. AMP-mediated defenses can enhance the stress response in adult flies and are regulated by the Imd pathway. To determine whether extracts of these four medicinal plants can reduce Drosophila intestinal damage, we analyzed AMP RNA levels (Dpt, Diptericin; Mtk, Metchnikowin) using real-time PCR. Slightly increased AMP levels in the experimental groups were observed compared with controls. Moreover, Dpt and Mtk RNA levels were dramatically increased in the C. pilosula feeding group 16 h after SDS treatment, with 40- and 23.5-fold increases, respectively, compared to the control group. The extracts of S. lappa, I. cylindrical var. major, and M. toosendan did not significantly affect AMP levels in the Drosophila gut. The RNA levels of other AMPs (AttA, Attacin A; CecC, Cecropin C; Dro3, Dromycin-like peptides 3; Dfn, Defencin) were similar between groups (data not shown). These results indicate that extracts of C. pilosula, S. lappa, I. cylindrical var. major, and M. toosendan can increase basal AMP levels, and C. pilosula extracts can also induce high levels of Dpt and Mtk 16 h after treatment with SDS in the Drosophila gut.

Medicinal plant extracts do not increase SDS-induced ISC proliferation in the midgut. After ingestion of toxic compounds, such as SDS or DSS, Drosophila ISCs increase their rate of proliferation in response to tissue damage. To analyze the protective effects of the four different medicinal plant extracts, the esg-Gal4 UAS-GFP marker (for ISCs and EBs) was used to assess adult flies after treatment with 0.6 % SDS. The numbers of ISCs and EBs were not significantly different between groups. This result indicates that these medicinal plant extracts do not induce stem cell proliferation in the Drosophila midgut in response to SDS.

Medicinal plant extracts are able to reduce SDS-induced cell death. In the Drosophila midgut, exposure to toxic compounds can increase death of epithelial cells. To determine whether the increased survival rate of adult flies resulted from decreased cell death in response to SDS, adult flies were treated with 0.6% SDS for 96 h. We observed a larger number of dead epithelial cells in the control group; however, flies fed extracts of traditional medicinal plants showed significantly reduced 7-AAD signals: 46.3%, 38.2%, 26.5%, and 54.4% when compared to control flies (100%), respectively. This result indicates that extracts of C. pilosula, S. lappa, I. cylindrical var. major, and M. toosendan can increase epithelial cell viability after toxic compound treatment.

Meicinal plant extracts have protective effects against SDS-induced gut damage and morphological changes. Previous reports showed that SDS is able to induce melanotic tumors and morphological changes in the Drosophila gut. After treatment with 0.6% SDS for 4 days, the guts of control flies appeared shorter than those of the experimental groups; furthermore, melanotic tumors were observed in the posterior midguts of control flies. In comparison, we observed significantly increased gut lengths in C. pilosula-, S. lappa-, I. cylindrical var. major-, and M. toosendan- fed groups: 40.3%, 55.4%, 41.5%, and 42.9% compared with control, respectively. In addition, we did not observe any melanotic masses in the experimental groups.

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