ABSTRACT
This study aims at making full use of microbial resources, and screening the active endophytic fungi of anti-rheumatoid arthritis from Zanthoxylum simulans. The endophytic fungi were cultured and isolated by tissue culture and scribing method, and the active strain of inhibiting the proliferation of human rheumatoid arthritis synovial fibroblasts (HFLS-RA) was screened by MTT method. Morphological characteristics and rDNA ITS1-5.8S-ITS2 sequences were applied for the taxonomy of endophytic fungi. Strains were isolated from Z. simulans. Among them, MK-05, MK-17, MK-19, MK-23 having inhibiting activity to HFLS-RA, the IC₅₀ were 0.367, 0.775, 0.689, 0.757 g·L⁻¹, respectively. By classic morphologic classification and sequencing the PCR-amplified rDNA ITS1-5.8S-ITS2 regions, four effective strains were identified as Botryosphaeria dothidea, Phomopsis sp., P. liquidambari and Diaporthe perseae. The active endophyic fungi that inhibited the proliferation of HFLS-RA were screened from Z. simulans for the first time, and the results lay the foundation for the development and utilization of the Z. simulans resources.
ABSTRACT
OBJECTIVE:To establish a method for the content determination of eudesmin in Zanthoxylum simulans. METH-ODS:HPLC was performed on the column of Kromasil C18 with mobile phase of acetonitrile-water(56:44,V/V)at flow rate of 1.0 ml/L,detection wavelength was 221 nm,column temperature was 25 ℃. RESULTS:The linear of eudesmin was 0.208-2.08 μg (r=0.999 8);RSDs of precision,reproducibility and stability tests were lower than 2.0%;recovery was 96.18%-103.04%(RSD=2.58,n=6). CONCLUSIONS:The method is simple and good reproducibility,and can be used for the content determination of eu-desmin in Z. simulans.