The Information Exchange Between Intestine And Major Organs Induced By Jujuboside A

Semen ziziphi Spinosae (SZS) is issued in the first batch of medical food by the nation.It showes various effects, such as nourishing heart, tonifying liver, tranquilizing mind,arresting sweating and promoting the secretion of saliva or body. It is mainly used to treatinsomnia, hyperhidrosis, body fluid deficiency, frighting palpitations and profusingdreaming. Modern pharmacological studies have demonstrated that Jujuboside A (JuA) isthe main bioactive component responsible for the sedative and hypnotic effects. However,the mechanism is still necessary to be investigated further. Intestinal tract is the vital organof the body, containing a variety of receptors and cytokines, plays a vital role in regulatingthe body’s immune response and transferring different bioinformations. In this research, thetissue distribution of JuA in rat was investigated. γ-aminobutyric acid (GABA) receptorgene expression induced by JuA in rat hippocampal cells was determined, and the signalexchange relationship between intestinal and various organs was analysed throughdetecting the changes of cytokines. This research would help to interpret the sedative andhypnotic mechanism of JuA.To investigate the tissue distribution of JuA, HPLC-MS/MS was applied to determinethe concentration of JuA in rats plasma and heart, liver, spleen, lung, kidney, stomach, brainintestine and muscle. The results showed that the compound distributed quickly in rats afterintravenous injected with2mg/kg of JuA. In the range of20-2000ng/mL, the linearity wassatisfying(r>0.9990). The extraction recoveries were all over80%, and the precision waslower than15%. The concentration was the highest in the intestine (2930.20±518.96ng/mL)，and lowest in the brain(144.27±22.31ng/mL). The order of the concentration was:kidney>spleen> liver>lung> stomach>heart>muscle. The concentrations decreased with theextension of time, falling to the lowest level in4hours, and there was no obvious specialsavings phenomenon in vivo.Hippocampus cells of rats were cultured in vitro. Through immunofluorescencetechnique, the cell purity was identifid with nerve cell markers of neuron tubulin-2(MAP-2). In addition, the effect of JuA on hippocampal cell vitality was investigatedthrough the MTT method. The results showed that the cells purity was perfect. After24h ofincubation with25μg/mL(P>0.05),50μg/mL(P<0.05) and100μg/mL(P<0.05) of JuA, thecell vitalities were all over100%compared with the blank control group.Through the Real-time Fluorescence Quantitative Polymerase Chain Reaction(RT-PCR), the effect of JuA on GABA receptor gene expression in rat hippocampal cellswas investigated. After48h incubation with JuA, the expression of GABAAα1, GABAAα5,GABABR1increased obviously (P<0.05), and GABAAβ1decreased (P<0.05) in the25μg/mL group. The expression of all the receptors increased in the50μg/mL group(P<0.05) and100μg/mLgroup (P<0.01). It indicated obvious dose-effect relationship between theJuA content and the effect. After24h, the expression of GABAAα1increased (P<0.01), andGABABR1、GABAAα5decreased in the50μg/mL group (P<0.05). The results demonstratedthe time-effect relationship in some degree. Compared with DZ, JuA showed stronger effecton the receptor mRNA expression, which indicated that JuA might have more speicfiedsedative hypnotic effect.With the liquid chip technology, the signal exchange relationship between intestineand various organs was investigated through detecting the cytokines in the differentbiosamples. According to the interaction among the source cells and function cells, thecytokine network was mapped to interpret the relationship of intestine and various organs.The results showed that the intestine secreted specific cytokines induced by JuA, and thecytokines would regulate the various organs to secrete their cytokines. According to thecytokine network, it indicated that intestine could effect the various with different strength:heart>spleen>brain>liver>lung>stomach>kidney. Cytokines are the important signalsbetween the intestine and organs communication, and cytokine network could provide aneffective means to investigate the interaction between the intestine and various organs.