Metabolomic study of raw and bran-fried Atractylodis Rhizoma on rats with spleen deficiency
Introduction
The spleen is one of the five internal organs of TCM. It bears the important physiological functions of human body transportation, ascending and dying, and plays an important role in the Tibetan elephant theory. Spleen deficiency is a weak syndrome caused by temper deficiencies and negligence in transportation [1]. Spleen deficiency means that the spleen does not disperse essence, neither can the fine substances be transported to the whole body, nor can it help the kidney to make water, while phlegm retention stagnate in the body, leading to abnormal blood lipid and blood sugar in the body [2].
Chinese Medicine was well known as its significance in the prevention and treatment of diseases, which is emphasized to recover the whole body and improve the resistance ability to disease through improving the internal environment of the human body [3]. Atractylodis Rhizoma is a commonly used Chinese herbal medicine in clinic. Its medicinal value was first recorded in Shennong Herbal Classic and listed as the top grade [4]. According to the 15th edition of the Chinese Pharmacopoeia, Atractylodis Rhizoma is the dry rhizome of Atractylodes lancea (Thunb.) DC. or Atractylodes chinensis (DC.) Koidz. It has been used as a Chinese medicine for more than 1500 years and has been mainly used for the treatment of drying dampness and strengthening spleen, dispelling wind and dispelling cold, and treating nyctalopia [5]. There are more than 20 kinds of processing methods of Atractylodis Rhizoma in the past dynasties, such as stir-fried coke, stir-fried yellow, stir-fried charcoal, bran, earthen, vinegar, salt and wine [6]. At present, other processed products except bran-fried products have been less used. Processing Chinese medicine can enhance the efficacy of Chinese medicine and change the trend of Chinese medicine action [7]. After the processing of Atractylodis Rhizoma, its spleen-strengthening effect increased and dryness decreased [8]. Prior to that, my research team has studied the difference between raw Atractylodis Rhizoma and bran-fried Atractylodis Rhizoma. It has found that the content of total volatile oil decreased after processing, and the chemical composition of volatile oil changed a lot, but not qualitatively. It was also found that bran stir-frying could improve the absorption and bioavailability of Atractyloside A, atractylenolide Ⅰ,Ⅱand Ⅲ in vivo. The urinary excretion kinetic parameters t1/2 and Ke of Atractyloside A, atractylenolide Ⅰ,Ⅱand Ⅲ had no significant changes, but the total excretion rate was significantly higher than that of raw products. And bran-fried Atractylodis Rhizoma has stronger effect on regulating spleen deficiency.
Metabolomics is a holistic analytical approach to the low-molecular-weight endogenous metabolites in various biological samples, and it can be used to monitor multiple metabolic pathways simultaneously and speculate the regulation of these pathways rationally based on the expression (up- or down-regulation) data of metabolites [9]. Metabolomic analysis techniques include LC–MS, GC–MS and NMR. Among those, LC–MS has become the most powerful tool. In particular, UPLC coupled with Q-TOF and MS has high resolution and sensitivity. It can extract polar metabolites and lipid metabolites at the same time. It can also realize large-scale qualitative and quantitative analysis of complex plasma metabolites [10]. Therefore, UHPLC-QTOF/MS combined with metabolomics analysis technology was used to identify the main metabolites related to diseases, and to understand the metabolic pathways of substances in body, so as to provide a theoretical basis for clarifying the scientific connotation of spleen deficiency syndrome [11]. The metabolomic studies on the processing mechanism of Chinese medicine have shown that different processed products of the same Chinese medicine can cause different responses to endogenous metabolites. However, there is no application of metabolomics technology to clarify the processing mechanism of Atractylodis Rhizoma. This paper is to study the processing mechanism of Atractylodis Rhizoma from the aspect of metabolomics.
In this study, UHPLC-QTOF/MS-based plasma metabolomics was applied to find metabolites and metabolic pathways associated with spleen deficiency. On this basis, the effects of raw Atractylodis Rhizoma and bran-fried Atractylodis Rhizoma on the same metabolites of spleen deficiency were compared, and the processing mechanism of Atractylodis Rhizoma was explained comprehensively from the perspective of metabolomics.
Section snippets
Reagents
Methanol and formic acid were purchased from Tianjin Kemiou Chemical Reagent Co., Ltd. (Tianjin, China). Purified water was obtained from Hangzhou Wahaha Group Co., Ltd. (Hangzhou, China) filtered through 0.22 μm filter membrane. Sennae Folium was provided by Herbal Chinese Medicine Decoction Co., Ltd. (Anhui, China) (License No. 140802), and the content of Sennoside A and sennoside B was in line with the standards of the Chinese Pharmacopoeia (2015). The ELISA kits were purchased from Shanghai
Behavioral comparison
Before the establishment of the model, the rats were completely healthy, without any abnormal signs. During the modeling period, the rats in control group had full mental state, white and dense fur, flexible movement and stool shape. However, the spleen deficiency rats presented pasty loose stools, weight loss, inactiveness, hypotrichotrophy, athrepsy, grouping, bad appetite, and lassitude. The spleen deficiency rats showed all the above signs,these signs are similar to the symptoms of TCM
Effect of Atractylodis Rhizoma on spleen deficiency
Spleen deficiency is a multifactorial etiological disease. There are several factors that play different roles in spleen deficiency, for example, digestion, absorption dysfunction membrane structure and function, and energy metabolism [16]. VIP, a vasoactive intestinal peptide, is a neurotransmitter that exists in the intestinal nervous system. VIP has the functions of vasodilation, anti-inflammatory immunomodulation, promotion of blood flow,cardiac contraction, lowering of blood pressure, and
Conclusion
In this study, both raw Atractylodis Rhizoma and bran-fried Atractylodis Rhizoma can Significant regulate the pathways of glucosamine metabolism, one carbon pool by folate, pantothenate and CoA biosynthesis and nicotinate and nicotinamide metabolism, and the effect of bran- fried Atractylodis Rhizoma on these pathways is better than that of raw Atractylodis Rhizoma. The present study could provide evidence that the mechanism of pharmacodynamic enhancement of the bran-fried Atractylodis Rhizoma
Authors contributions
Beixue Zhang, Xiaojie Qi and Qian Cai conceived and designed the experiments; Beixue Zhang and Xiaojie Qi performed the experiments; Beixue Zhang analyzed the data; Beixue Zhang wrote the paper.
Declaration of Competing Interest
We have no conflict of interest to declare.
Acknowledgements
This research work was financially supported by grants from the National Natural Science Foundation of China (No: 81202919) and the National Administration of Traditional Chinese Medicine (State Administration of Traditional Chinese Medicine) Chinese Medicine Industry Research Project (No: 20110700712).
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