Renoprotective Effects of Total Glucosides from Paeony against Nephrotoxicity Induced by Total Alkaloids from Semen Strychni

Hindawi Evidence-Based Complementary and Alternative Medicine Volume 2017, Article ID 8256278, 9 pages https://doi.org/10.1155/2017/8256278 Research Article Renoprotective Effects of Total Glucosides from Paeony against Nephrotoxicity Induced by Total Alkaloids from Semen Strychni Mingming Lv,1 Meiyu Zhang,2 Yezhe Cheng,2 Kexia Zhang,2 Chenzhi Hou,1 and Xiaohui Chen1 1 School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016, China 2 Correspondence should be addressed to Xiaohui Chen; cxh Received 1 May 2017; Revised 18 July 2017; Accepted 2 August 2017; Published 22 October 2017 Academic Editor: Caigan Du Copyright © 2017 Mingming Lv et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Semen Strychni have been shown to have therapeutic effect in improving blood circulation, relieving rheumatic pain, and treating cancer. However, Semen Strychni could cause severe nephrotoxicity. The present study was designed to evaluate whether treatment with total glucosides from paeony (TGP) has renoprotective effect against nephrotoxicity induced by total alkaloids from Semen Strychni (TAS). The levels of blood urea nitrogen (BUN) and creatinine (Cr) were determined and histopathological changes were also examined to evaluate renal injury. Moreover, a HPLC-MS method was developed and validated to investigate the comparative toxicokinetics of strychnine and brucine in rats plasma after oral administration of TAS and pretreatment with TGP. Results demonstrated that the levels of BUN and Cr were significantly increased (𝑝 < 0.05) in TAS group, together with tubule epithelium cloudy swelling, degeneration, and glomerular atrophy in rats’ kidneys. The TAS-induced kidney damage was alleviated after pretreatment with TGP. Besides, 𝑇max of strychnine and brucine were increased and 𝑇1/2 of strychnine and brucine were decreased after pretreatment with TGP. The toxicokinetics study showed that pretreatment with TGP could attenuate the absorption of strychnine and brucine, as well as accelerate their elimination. These results suggest that TGP possesses renoprotective effects. 1. Introduction Semen Strychni is a traditional Chinese herb, mainly used to treat rheumatoid arthritis, nervous disease, and traumatic pains and to promote blood circulation and remove blood stasis [1–3]. Total alkaloid from Semen Strychni (TAS) is a concentrated bioactive extract of Semen Strychni, which mainly consists of strychnine and brucine [4, 5]. Strychnine and brucine possess analgesic, anti-inflammatory, and antitumor effects as well as toxicity [6, 7]. The use of Semen Strychni was limited for its extreme toxicity, and it can also cause cardiac arrest, epileptic seizures, and nephrotoxicity [8, 9]. The suggested therapeutic dose of Semen Strychni is 0.3–0.6 mg/per day and the dose of its prime toxic ingredientstrychnine is 5–10 mg [10–12]. In previous study, Semen Strychni has obvious renal injury [13]. Thus, it is necessary to reduce the nephrotoxicity of Semen Strychni. Some research showed that combining Semen Strychni with Paeonia lactiflora could reduce the content of strychnine and brucine, increase the therapeutic effect, and reduce the toxicity of Semen Strychni [14, 15]. Semen Strychni and Paeonia lactiflora have been compatibly used in many proprietary Chinese medicines such as Gujinwan Capsule, Huoxuezhitong Paste, and Huoluozhenfeng Pill to treat osteopenia and arthralgia and relieve pain. Total glucosides of paeony (TGP) are the predominate effective ingredients of the traditional Chinese herb, Paeonia alba radix (roots of Paeonia lactiflora Pall.), which mainly contains paeoniflorin and albiflorin. It was reported that TGP possesses analgesic, anti-inflammatory, and immunoregulatory effects [16, 17] and it was exerted renoprotection with antioxidative injury, preventing renal tubulointerstitial injury and blocking TLRs action [18, 19]. It was also reported that TGP plays therapeutic role in experimental diabetic nephropathy [10]; moreover, 2 Evidence-Based Complementary and Alternative Medicine O OC（3 OH OH N N H H O （3 CO H H H N H O （3 CO O H H N N OC（3 O H H O O OH O O OH O O (a) (b) (c) Figure 1: Chemical structures of strychnine (a), brucine (b), and IS clarithromycin (c). TGP has already been used to treat chronic nephritis in clinic [20]. As far as we know, few investigation were developed to verify the effects of TGP on nephrotoxicity induced by TAS. The present study was designed to evaluate whether treatment with total glucosides from paeony exerts renoprotective effect against nephrotoxicity induced by total alkaloids from Semen Strychni. The levels of serum BUN, Cr, and kidney histopathological changes were used to certify the nephrotoxicity of TAS. What is more, the antidotal effects of TGP on the toxicokinetics of strychnine and brucine in rats were studied. 2. Materials and Methods 2.1. Material and Regents. Semen Strychni, Paeonia lactiflora Pall, and Aristolochia manshuriensis (AM) were purchased from Anguo Lengbei Co., Ltd (Hebei, China). All herbs were authenticated by Professor Ying Jia (Pharmacognosy Department, Shenyang Pharmaceutical University). The reference standards of strychnine and brucine were obtained from National Institute for control of Pharmaceutical and Biological Products (Beijing, China). Clarithromycin as internal standard (IS) was provided by Sigma-Aldrich (MO, USA) (Figure 1). The purity of each reference standard was all above 98%. HPLC grade methanol and acetonitrile were purchased from Fisher Scientific (Nanjing, China). HPLC Glacial acetic acid was obtained from Yuwang Industry Co. Ltd. (Shandong, China). Distilled water, prepared from demineralized water, was used throughout experiment. 2.2. Herb Preparation. The dried seeds of Semen Strychni (100 g) were extracted three times by refluxing with 1 L of 70% ethanol (1 : 10 m/v) for 1 h each time. The collected solvent was concentrated under rotary evaporation to remove all ethanol. The residue was suspended into water and acidified to pH = 2 with HCl (1.0 mol/L) and then extracted with methylene chloride. The pH of aqueous layer was adjusted to 12 with 4.0 mol/L NaOH and then extracted with methylene chloride once more. The methylene chloride layer was concentrated under rotary evaporation to remove all ethanol, and the resultant residue was the TAS extract. The concentrations of strychnine and brucine were determined to be 33.1% and 25.9% by HPLC method. The powder of Paeonia lactiflora Pall radix (250 g) was extracted three times by refluxing with 2 L of 70% ethanol (1 : 8, m/v) for 2 h each time. The filtrates were concentrated by removing all ethanol solvent in rotary evapo (...truncated)