The exosome-encapsulated compounds (Exo-Bai, Exo-Hed, Exo-Nef, Exo-Rg3, Exo-Rapa and Exo-Rb1) were prepared by the RT method, as described [22]. In brief, 100 μg Bai, Hed, Nef, Rg3 and Rb1 (purchased from Sichuan Weikeqi Biological Technology Co., Ltd., Sichuan, China) or Rapa (purchased from Guangzhou Tomums Life Science Co., Ltd., Guangdong, China) were incubated with exosomes (25 μg) in a ratio of 4:1 in PBS at 22 °C for 18 h before being subjected to quantitation.

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.

The administered dose of AlCl3

(Aladdin, Shanghai, China) was previously reported to exert a change

in bone activity.11 Rg3 (purity >98%; Weikeqi, Sichuan, China) was

dissolved in isotonic physiological saline (0.9% NaCl) and

administered for 30 days at a dose previously reported to influence

the bone activity

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).

AlCl3 (Aladdin, Shanghai, China) was administered in accordance with the dose previously reported for exerting a change in bone activity for 120 days [25]. Rg3 (Weikeqi, Sichuan, China) was dissolved in isotonic physiological saline (0.9% NaCl), and a dose administered in accordance with that previously reported as exerting an influence on bone, for 30 days [24].

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

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.

The ginsenoside Rb3 (Rb3) was isolated from SCLPN, and its structure was characterized by spectroscopic analysis with both ESI-MS and 13C NMR. 20(S)-Ginsenoside Rg3 (Rg3, Lot No.: 14,197–60-5-15-1), 20(S)-ginsenoside Rh2 (Rh2, Lot No.: 78214-33-2-14-6), compound K (C-K, Lot No.: 39262-14-1-14-9) and 20(S)-protopanaxadiol (PPD, Lot No.: 30636-90-9-14-2) were supplied by Sichuan Weikeqi Biological Technology Co. Ltd. (Chengdu, China). These compounds are white powders with a purity of 98%, which was verified by high

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.