| Insomnia is a serious chronic disease.According to WHO statistics,the global sleep disorder rate is as high as 1/3.In China,more than 300 million people have sleep disorders,and the number of people is still rising year by year.Medical treatment is the first choice of current treatment guidelines.Unfortunately,most of the chemicals used in western medicine for insomnia are addictive.It will compensate for rebound after they stopped taking the medicine,and withdrawal symptoms are so obvious that are easy to produce dependence and resistance to drugs.What’s worse is that they need to be taken for a long time or for a lifetime.The curative effect and function of traditional Chinese medicine in the prevention and treatment of chronic diseases(insomnia,cardiovascular and cerebrovascular diseases,diabetes,etc.)have been paid more attention by pharmacists.Losing sleep is called insomnia in traditional Chinese medicine.There are many famous prescriptions about the syndrome differentiation and treatment of insomnia.Most of traditional Chinese medicine are mild,safe and cheap.Ailanthi Cortex is the dry root bark or dry bark from tree of heaven(Ailanthus altissima(Mill)Swingle)in the family of Simaroubaceae.The treatment of insomnia,epilepsy and palpitation by Ailanthi Cortex has been recorded in famous medical works of all ages.As early as in Tang Dynasty,Ailanthi Cortex was recorded to be used as treat mental illness.According to compendium of Materia Medica written by ancient book of Ming Dynasty,the decoction of Ailanthus altissima’s leaf and root bark had the effects of sedation and hypnosis.In recent years,Korea,Japan and other easten countries have done a lot of research on the anti-inflammatory,antibacterial and anti-virus pharmacological effects of the root bark of Ailanthus altissima,but there is no report on the sedative and hypnotic effects,the material basis of its efficacy is still unclear,and the mechanism of its pharmacological action is unclear.Modern research on the root bark of Ailanthus altissima has found that its main chemical component is alkaloid,and it is also an active component of anti-inflammatory,antibacterial and antiviral effects.Therefore,using modern scientific means to carry out a more systematic study of pharmacological action,chemical composition and preliminary mechanism of action of monomer compounds,to confirm whether it has sedative and hypnotic effect and whether it is the main effective substance of Ailanthi Cortex,which has a very important scientific research significance for the inheritance and expansion of the efficacy of traditional Chinese medicine of Ailanthi Cortex.By heating and refluxing with 85% ethanol,we get 470.18 g total extract and 160.25 g total alkaloids by traditional methods of acid extraction and alkali precipitation.The content of alkaloid in total alkaloid was 51% by acid dye colorimetry.The sedative and hypnotic effects of total extracts and alkaloids from Ailanthi Cortex were studied by behavioral observation.The sedative and hypnotic activities of these two extracts were investigated by observing the parameters of mice’s autonomous activities and standing times,and the sleep latency and sleep prolongation time of mice assisted by pentobarbital sodium.The results showed that: compared with the blank group,the high-dose group of total extract of Ailanthi Cortex could significantly reduce the activity times of mice(P < 0.05).There was no significant difference among the low,middle and high dose groups.The middle and high dose groups could significantly shorten the sleep latency of mice(P < 0.05),and the high dose group could prolong the sleep time of mice(P < 0.05).Compared with the blank group,the middle and high dose of total alkaloids could reduce the number of autonomous activities and standing of mice(P < 0.05),the high dose group could shorten the sleep latency of mice(P < 0.05),and the low,middle and high dose groups could prolong the sleep time of mice(P < 0.05).The results showed that the total alkaloid and total extract from Ailanthi Cortex had certain sedative and hypnotic activity,and the total alkaloid was the active part of the total extract of Ailanthi Cortex.We studied the chemical constituents of total alkaloids from Ailanthi Cortex by silica gel column chromatography and Sephadex LH-20 gel column chromatography,ODS reversed phase column chromatography and semi high performance liquid chromatography(HPLC).The structures of 14 compounds were identified by means of physicochemical constants and spectral data analysis,including six β-carboline type alkaloids,namely,β-carboline(1),3-ethyl-β-carboline(2),5-methoxy-3-ethy-β-carboline(3),14,15-dihydroxy-5-methoxy-β-carboline(4),11-methoxy-3-methyl-β-carboline(5)and 3-methoxycarbonyl-β-carboline(6);Four canthin-6-one type alkaloids,namely canthin-6-one(7),4,5-dimethoxycanthin-6-one(8),4-methoxy-5-hydroxycanthin-6-one(9)and 11-hydroxycanthin-6-one(10);One tirucallane-type triterpenoid,namely piscidinol A(11);Two secodammar-type triterpenoids,namely 12β,20(S),24(R)-trihydroxydammar-25-ene-3-one(12)and 12β,20(S),25-trihydroxydammar-23-ene-3-one(13);One pennogenin-type steroidal saponin,namely 3-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside(14).Among them,alkaloids 3 and 4 are new compounds.Compounds 12～14 are isolated from the plants of the Simaroubaceae for the first time.In this study,pennogenin-type steroidal saponins are isolated from the plants of the Simaroubaceae for the first time.MTT method was used to investigate the effect of two new alkaloids and eight known alkaloids on the survival rate of adrenal chromaffin tumor cells(PC12)in serum-free rats,The results showed that compound 7 and 8 increased the cell survival rate to 77.5 ± 2.3%(P < 0.05)and 70.0 ± 2.3% compared with the serum-free model group,showing a good neuroprotective effect.Compounds 1～6,9 and 10,compared with the serum-free model group,reduced the cell survival rate,indicating that they may have some damage to the cells.The best protective compound 7 was selected to study its sedative and hypnotic effects and mechanism by behavioral observation and Enzyme-linked immunosorbent assay(ELISA).The results showed that the middle and high dose group of compound 7 had the effects of reducing the number of activities and standing,shortening the sleep latency and prolonging the sleep time(P < 0.05),Each dose group can improve the incidence of sleep(P < 0.05);the middle and high dose groups can reduce the content of 5-Hydroxytryptamine(5-HT)and noradrenaline(NE)in the brain,and increase the content of γ-Aminobutyric acid(GABA)(P< 0.05).It is suggested that the compound has obvious synergistic effect on improving sleep,and its mechanism may be closely related to the regulation of the expression of related neurotransmitters(5-HT,NE,GABA).Innovation and significance of the project:1.In this study,the sedative and hypnotic activities from Ailanthi Cortex extract were studied systematically for the first time,and the pharmacological activities and mechanism of the representative compounds with sedative and hypnotic effects in Ailanthi Cortex were studied by behavior observation and ELISA for the first time.2.In this paper,the chemical constituents of total alkaloids of Ailanthi Cortex with sedative and hypnotic effects were studied systematically,14 compounds were isolated and identified,and 2 new compounds were found.Among these constituents,including the known β-carboline-type alkaloids and canthin-6-one-type alkaloids.Two secodammar-type triterpenoids and one pennogenin-type steroidal saponin were isolated from the plants of the Simaroubaceae for the first time.This is the first time that pennogenin-type steroidal saponins have been found from the plants of the Simaroubaceae.3.The preliminary pharmacological research on the effective part of Ailanthi Cortex and the systematic chemical composition research in this study clarified the material Ailanthi Cortex sedation,which was not only improve the pharmacological activity and chemical composition data of Ailanthi Cortex,but also can be provided as the scientific basis and theory for the further development and utilization of Ailanthi Cortex. |
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