The Discovery Of Ginger Charcoal Nano-components And Its Analgesic Effects

Background: Zingiberis Rhizome Carbonisata,one of a Chinese charcoal drugs,is the dry roots of dried ginger(Zingiber officinale Rosc.)after high temperature processing products.As widely used in the theory of typhoid miscellaneous diseases,dried ginger has the effects of warming spleen and stomach for dispelling cold,rescuing from collapse by restoring yang and dry wet phlegm elimination,which can be applied to treat cold pain in the abdomen,vomiting,diarrhea,cold limbs,weak pulse,phlegm retention and asthmatic cough.After processing,the effect of charcoal ginger tend to moderate,which can be applied for warming channel for arresting bleeding,antidiarrheal,warming spleen and stomach and relieve pain.For pain,both dried ginger and Zingiberis Rhizome Carbonisata have good effects.However,the effective ingredients and mechanism of action of Zingiberis Rhizome Carbonisata are still not clear at present.In this study,the analgesic effect and material basis of Zingiberis Rhizome Carbonisata were investgated by the modern nanotechnology assay.Objectives:1.Nanoscale components of ginger carbon were isolated,and the effective components were screened and identified.2.Optimize the preparation temperature and time of charcoal ginger.3.Development of standardized characterization procedure of carbon ginger nanoscale components.4.Evaluate the analgesic effect and explore the analgesic mechanism by studying the cytotoxicity of carbon ginger nano-components.Methods:1.Preparation and purification of charcoal ginger.Using hot plate experiment in mice,the effective parts of charcoal ginger for analgesia were screened.High performance liquid phase method(HPLC)was used to compare the fingerprint of dry ginger,charcoal ginger and the inner and outer parts of ginger charcoal dialysis bag,and nanotechnology was used to identify the effective parts.2.Preparate the charcoal ginger in different temperature and time.A series of methods,including high resolution electron microscopy(HR-TEM),X-ray photoelectron spectroscopy(XPS),ultraviolet-visible spectrophotometer(UV vis)and Fourier infrared spectrum(FT IR),was used for characterization of different batches charcoal gingers and optimize the preparation conditions.3.RAW264.7 macrophage and cck-8 proliferation assay were used to determine the cytotoxicity of carbon ginger nano-components and investgate the safe dose.4.Three dosages of high,medium and low were set for the carbon ginger nano-components prepared under the optimized conditions,and the analgesic effect was further investigated by hot plate test in mice and hot bath tail shaking test in mice,and the appropriate dosage was explored.5.The effects of naloxone on the analgesic effect of charcoal ginger nano-components were evaluated.To preliminarily study,the effects of ginger charcoal nano-components on enkephalin,β-endorphin,and 5-hydroxytryptamine in rats were detected,Results:1.The effective analgesic parts of charcoal ginger were selected by hot plate test in mice.The nano-components in the dialysis bag,morphine group and ginger charcoal water extract group showed the significant analgesic effects.HPLC spectrum showed that there was no peak of small molecule compounds in the dialysis bag.The solution in the dialysis bag was characterized and identified as speckle substance with particle size of about 0.23 nm,which named nanoscale component of charcoal ginger,also plays an analgesic role.2.The charcoal ginger were preparated of the different temperature under the condition of different time,by high temperature pyrolysis method.Through spectral analysis,combined with charcoal production rate,extraction rate,and the mouse hot plate test,the analgesic effect of charcoal gingers were measured.And the 350 ℃,1 h was confirmed of the optimized charcoal condition.The particle size was 1.5-5.5nm and the lattice spacing was 0.23 nm.At about 275 nm,it has the maximum ultraviolet absorption.The maximum excitation wavelength in the fluorescence spectrum was 375 nm,and the corresponding maximum emission wavelength was 469 nm.It mainly contains C,O and N elements,and the main functional groups on its surface are hydroxyl group,carboxyl group and amino group.3.Under the optimal preparation conditions,the safe dose of carbon ginger nano-components was below 3000 g/mL.4.In hot plate experiment and tail shaking experiment,all of high,medium and low doses nanoscale conponents have significant analgesic effect(P < 0.01),and the analgesic effect is more obvious when the dose is low and medium.5.The effects of naloxone,a morphine antagonist,on the analgesic effect of carbon ginger nano-components were determined.The effects of ginger charcoal nano-components on enkephalin,β-endorphin and 5-hydroxytryptamine in rats were detected,and the analgesic mechanism was studied.Conclusion:In this study,we isolated nanoscale component from charcoal ginger for the first time.And the charcoal ginger preparation temperature and time were optimized as 350 ℃and 1 hour.Nano-components were detailed characterized.Our results breakthrough the existing concept and current method on traditional Chinese medicine material base.On this basis,we found that the nano-component of charcoal ginger has high safety and analgesic effect,and its analgesic mechanism is central analgesia.This study is the first to find new ingredients in charcoal ginger,and explore the efficacy of this ingredient from a new perspective.We also provide a new view and method for the further study of other charcoal drugs.