N butanol

Crude drug samples were collected in Ganchuan town, Xingan County, Jiangxi Province, China, which were later authenticated as Euodia rutaecarpa (Juss.) Benth. by Lan Cao, an associate professor in Jiangxi University of TCM (JXUTCM). Voucher specimens are preserved in the Herbarium of Pharmacognosy, School of Pharmaceutical Sciences, JXUTCM. Analytical-grade chloroform, ethyl acetate, and n-butanol were all purchased from Xilong Chemical Co., Ltd. (Guangzhou, Guangdong Province, China); ALT and AST kit were purchased from Nanchang Biotech A and C Biotechnical Industry Inc. (Nanchang, Jiangxi Province, China); chromatographic-grade methanol and acetonitrile were purchased from TEDIA (Fairfield, OH, USA); and ultrapure water was prepared by a Milli-Q water purification system (Millipore, Bedford, MA, USA).
SPF-grade Kunming mice with weights ranging from 18 to 22 g, half male and half female, were provided by Hunan SJA Laboratory Animal Co., Ltd. (approval No. SCXK(Xiang)2011-0003. Changsha, Hunan Province, China). The Experimental Animal Ethics Committee of Jiangxi University of TCM approved all animal protocols. The animal experiments were carried out according to the European Community guidelines for the use of experimental animals.

Zheshen No. 2 P. Heterophylla (Miq.) Pax Chinese herbal was purchased from Ningde Nanling Agricultural Co., Ltd. Zherong County of the Ningde region of the Fujian Province in China is the advocate producing area of TZS. The TZS was cut into pieces and then dried below 60°C.
Chromatographic pure methanol was purchased from Merck KGaA (Darmstadt, Germany). Chromatographic pure acetonitrile was purchased from Fisher Scientific UK Ltd. (Leicestershire, England). Analytical grade reagents including petroleum ether, ethyl acetate, ethanol, n-butanol, ammonia, and chloroform were bought from Xilong Scientific Co., Ltd. (Shantou, China). Interleukin-10 (IL-10) and tumor necrosis factor (TNF-α) enzyme linked immunosorbent assay (ELISA) kits were purchased from cloud-clone corp.

The roots and rhizomes of AGS were purchased from Guangzhou Zhixin medicinal herbs Co., Ltd (Lot number: 20,191,201, Guangdong, China) and identified by Professor Suying Tian (Guangdong Pharmaceutical University, Guangzhou, China). The ethanol, petroleum ether (30–60 ℃, 60–90 ℃), ethyl acetate, and n-butanol were obtained from Xilong Chemical Co., Ltd (Guangdong, China). Diatomite was obtained from Dingshengxin Chemical Co., Ltd (Tianjin, China). McCoy’s 5 A medium was purchased from Procell Life Science&Technology Co., Ltd (Wuhan, China). Leibovitz’s L-15 medium was purchased from Dalian Meilun Biotechnology Co., Ltd (Dalian, China). Fetal bovine serum, phosphate-buffered saline, 5% trypsin-EDTA were purchased from Gibco (Carlsbad, CA, United States). Penicillin-streptomycin (P/S) and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) were purchased from Beyotime Biotechnology (Shanghai, China).