人參皂苷Rd/人參皂甙RD/絞股藍(lán)皂苷VIII/(2S,3R,4S,5S,6R)-2-(((2R,3R,4S,5S,6R)-4,5-二羥基-2-(((3S,5R,8R,9R,10R,12R,13R,14R,17S)-12-羥基-4,4,8,10,14-五甲基-17-((S)-6-甲基-2-(((2S,3R,4S,5S,6R)-3,4,5-三羥基-6-(羥甲基)四氫-2H-吡喃-2-基)氧基)庚-5-烯-2-基)十六氫-1H-環(huán)戊烷[a]菲-3-基)氧基)-6-(羥甲基)四氫-2H-吡喃-3-基)氧基)-6-(羥甲基)四氫-2H-吡喃-3,4,5-三醇/絞股藍(lán)總皂甙

The reference standards of notoginsenoside R1, ginsenosides Rg1, Re, Rb1, Rc, 20(S)–Rh1, 20(R)-Rh1, Rd, Rg6, F4, Rk3, Rh4, 20(S)-Rg3, 20(R)-Rg3, Rk1, and Rg5, all with a purity ≥98%, were purchased from Sichuan Weikeqi Biological Technology Co., Ltd. (Chengdu, Sichuan, China). EPO was purchased from Chengdu Di'ao Jiuhong Pharmaceutical Factory (Chengdu, Sichuan, China). FEJ was purchased from Shandong Dong'e E'jiao Co., Ltd.

Liquiritin apioside, liquiritin, neoliquiritin, isoliquiritin apioside, isoliquiritin, ononin, naringin, liquiritigenin, poricoic acid A, poricoic acid B, poricoic acid C, glycyrrhetinic acid, dehydropachymic acid, pachymic acid, ginsenoside Re, ginsenoside Rg1, ginsenoside Rf, isoliquiritigenin, ginsenoside Rb1, ginsenoside Rg2, ginsenoside Rg3, ginsenoside Rg5, 20(S)-ginsenoside Rh1, notoginsenoside R1, ginsenoside Ro, ginsenoside Rc, formononetin, ginsenoside Rb2, glycyrrhizic acid, ginsenoside Rd, ginsenoside Rk1, and atractylenolide III were purchased from Chengdu DeSiTe Biological Technology Co., Ltd. (Chengdu, China) or Sichuan Weikeqi Biological Technology Co., Ltd. (Chengdu, China).

P. notoginseng were purchased from Wenshan County of Yunnan Province. P. notoginseng is dried by oven, powdered by a disintegrator (HX-200A, Tianjin Taisite Instrument Co., LTD, China) and sieved 40 mesh. The standard of saponins Rb1 (HPLC ≥ 98%), Rd (HPLC ≥ 98%) and Rg3 (HPLC ≥ 98%) were purchased from weikeqi-biotechnology limited company of Sichuan. Methanol and acetonitrile were of HPLC grade (J & K Chemical Ltd., China). Ethanol of analytical grade for extraction was bought from

3′-Hydroxypuerarin, puerarin, 3′-methoxypuerarin, mirificin, daidzin and daidzein were obtained from Chengdu Purechem-standard Co. Ginsenoside Rg1, ginsenoside Rh1, ginsenoside Rd, astragaloside IV and 20(S)-ginsenoside Rg3 were purchased from Chendu Herbpurify Co. Astragalin was obtained from Sichuan Weikeqi Biotechnology Co. Linoleic acid and palmitic acid were purchased from Shanghai Ryon Biological Technology Co. HPLC-grade acetonitrile was obtained from Merck Co.

Penicillin (200,000 units) was intramuscularly injected every day to protect from post-operative infection. A twice-a-day manual bladder expression was performed until spontaneous voiding returned. Rats in the GS Rd groups received an intraperitoneal injection of GS Rd (purity: HPLC ≥98%, WKQ-00514; Sichuan Weikeqi Biological Technology Co., Ltd, Chengdu, China), and rats in the DEX group received an intraperitoneal injection of DEX (Tianjin Xinzheng, Tianjin, China) 1 hr before the surgery and once daily for 14 days. Rats in the sham and SCI groups received 0.9% NaCl only.

Oleanolic acid, maslinic acid, hederagenin, ursolic acid, corosolic acid, asiatic acid, β-boswellic acid, glycyrrhetic acid, celastrol, betulin, betulinicaldehyde, betulinic acid, pachymic acid, ganoderic acid B, polygalacic acid, glycyrrhizic acid, lupeol, ginsenosideol F1, ginsenoside Rg1, ginsenoside Rg2, were purchased from Dalian Meilun Company (Beijing, China), ginsenoside Rd, ginsenoside R1, Notoginsenoside R1 were purchased from Sichuan Weikeqi Biotechnology Co., Ltd. (Chengdu, China), ginsenoside Re, ginsenoside Rh1 were purchased from Chengdu Pfeid Biotech Technology Co., Ltd. (Chengdu, China), ginsenoside Ro, ginsenoside Rh4, and ginsenoside F4 were purchased from Chenguang Bio (Handan, China).

Isomaltose was purchased from Tokyo Chemical Industry (Tokyo, Japan). Protopanaxatriol and protopanaxadiol were obtained from ChromaDex (Irvine, CA). Compound K, ginsenosides Rh1, Rd, F1, F2, Rg3, and Rh2 were obtained from Sichuan Weikeqi Biological Technology (Chengdu, Sichuam, China). Lectin from Phytolacca americana (pokeweed mitogen, PWM), dextran standards certified according to Deutsches Institut für Normung (molecular weight; 668 kDa), Dulbecco’s modified Eagle’s medium

Qualitative and quantitative analyses of SPN were performed as previous report (Xiong et al., 2017a). In brief, notoginsenoside R1 and ginsenosides Rg1, Re, 20(R)-Rh1, Rb1, Rd, Rk3, Rh4, 20(S)-Rg3, 20(R)-Rg3 (Sichuan Weikeqi Biological Technology Co., Ltd. Sichuan, China) with purity ≧ 98% were used as standard compounds. The analysis was performed on an Agilent 1260HPLC system (Agilent Technologies) equipped with a G1311B Pump, a G4212B DAD detector, and a G1329B autosampler.

Recombinant human UGT isoforms (UGT1A1, UGT1A6, UGT1A7, UGT1A8, UGT1A9, UGT1A10, UGT2B7, UGT2B15) expressed in baculovirus-infected insect cells were obtained from BD Gentest Corp. (Woburn, MA, USA). Ginsenosides Rb1, Rb2, Rc, Rd, Rg3, Rh2, C-K, ppd, ppt, Re, Rg1, Rh1, F1, and F2 were purchased from Sichuan Weikeqi Biotechnology Company (Chengdu, Sichuan, China). The purity of these compounds was above 95%. All other reagents were of HPLC grade or of the highest grade commercially available.

Acetonitrile of chromatographic grade and ferrous chloride and hydrogen peroxide of analytical grade were purchased from Merck Chemical Co. (Darmstadt, Germany). Notoginsenoside R1, ginsenosides Rg1, Re, Rb1, Rd, Rh1, Rk3, Rh4, 20(S)-Rg3, and 20(R)-Rg3 (Sichuan Weikeqi Biological Technology Co., Ltd. Chengdu, China) with a purity ≥ 98% were used as the standard compounds.

The water layer was further partitioned with water-saturated BuOH three times, yielding the BuOH and water fractions (0.20 and 4.4 g, respectively). Ginsenosides F1, F2, Rd, Rg3, Rh1, Rh2, protopanaxadiol, and protopanaxatriol were obtained from Sichuan Weikeqi Biological Technology (Chengdu, Sichuan, China); ginsenosides Rg1 and Rb1 were obtained from Wako Pure Chemical Industries (Osaka, Japan)

Ginsenosides Rg1 and Rb1 were obtained from Wako Pure Reagents (Osaka). Compound K was obtained from ChromaDex (Irvine, CA, USA). Ginsenosides F1, F2, Rd, Rg3, Rh1, Rh2, protopanaxadiol and protopanaxatriol were obtained from Sichuan Weikeqi Biological Technology (Chengdu, Sichuam, China). Chemical structures of ginsenosides used in the present study are shown in Fig. 1.

Ginsenosides Rg1 and Rb1 were obtained from Wako Pure Reagents (Osaka, Japan). Compound K was obtained from ChromaDex (Irvine, CA). Ginsenosides Rd, Rh1, F1, and F2 were obtained from Sichuan Weikeqi Biological Technology (Chengdu, Sichuam, China). The chemical structures of these ginsenosides are shown in Fig. 2.