Rhizoma Anemarrhenae-cortex Phel Lodendri Medicine Effective Parts Of Quality Research And Its Effect On The Pharmacodynamics Research

ObjectiveThe medicine suit,and said:“matched pair of medicine"?"pair" or"sister drug",is under the guidance of the basic theory of traditional Chinese medicine(TCM)treatment based on syndrome differentiation and drug compatibility theory as the foundation,to the etiology and pathogenesi s of disease and syndrome characteristics as the basis,habits or often clinically coordinating related groups of pairs of medicine.The drug coup le is the basest?most foundational and simplest form in the prescription of TCM.As the basis of compatibility of TCM and a special phenomenon,centrally reflects the general rule of compatibility of traditional Chinese medicine theory:Similar?relativity?supplemented?phase system,on the contrary.Rhizoma Anemarrhenae and Cortex Phellodendri is a common medicine pair for classic.The earliest appearance of this couple is from?LAN shi mi cang?,which is said gate-freeing pill,and named Zishen Pellet,written by Li GAO in Jing-dynasty.Have shown in many classical prescriptions,such as Zishen pill clearance?Zhibaidihuang Pill?Large Yin Nourishing Pill?Du Zhong Pill?Tai ji pills?Hidden Tiger Pill?et al.According to the theory of traditioal Chinese medicine,Rhizoma anemarrhenae,bitter cold-natured,owned by the kidney?lung?stomach sutra,is one of clearing virtual hot medicine which is very commonly used in clinical.Cortex Phellodendri,bitter in taste and cold-natured,owned by the kidney?bladder?large intestinal meridian,is with strong Yin,clear heat dry wet,purging fire detoxification efficacy.Now the pharmacological research have showed,The Saponins,flavonoids and polysaccharides et al,contained in anemarrhena,composition have obvious hypoglycemic,reducing blood fat or anti-inflammatory effects.while berberine in cortex phellodendris total alkaloid is the main ingredient which can improve renal dysfunction in diabetic nephropathy rats,has obvious anti diabetes,reducing blood lipid,anti-inflammatory and immune regulation,and so on,improve the diabetic peripheral neuropathy in patients with nerve conduction velocity has a positive role.Methods1.TO study the methods for effective components of the content of Rhizoma anemarrhenae-cortex phellodendri medicine herb pairs To develop a new method using UV?HPLC and HPLC-ELSD,using berberine hydrochloride,cortex phellodendri,rhizoma anemarrhenae saponins BII and mangifer-in as controls,for determination content of Total alkaloid,total saponins,flavonoids,and berberine,cortex phellodendri,alkali,rhizoma anemarrhenae saponins BII,mangiferin in the extractive from Rhizoma anemarrhenae-cortex phellodendri medicine.2.The process research of herb pairs extraction,purification,concentration technologyThis research further improved the extraction,separation and enrichment process of Rhizoma anemarrhenae-cortex phellodendri.Through the underbrush,to transfer rate of target component as the index,established the optimal extraction process,the PIPO-00 macroporous resin column with 732 cation exchange resin column as the purification,double column in series,using high pressure on a sample of adsorption and elution high-pressure,the total saponins,total alkaloids,flavonoids,purification of enrichment.3.Rhizoma anemarrhenae cortex phellodendri total extract the pharmacody-namics study of diabetic peripheral neuropathy in mice200 male C57BL/6j mice,20 in each group,divided into normal group,model group,metformin group,xuezhikang group:total saponins of high/low dose group,high/low dosage group of total alkaloid,parker always extract high/low dose group.In addition to the normal group,other groups induced form of diabetes model in mice by intraperitoneal injection of STZ and high fat feed.Lavage once,every morning for six months(24 weeks).Respectively for 8,16 and 24 weeks,determiningn of serum GLU,TC,TG,LDL-C,INS,UA,T-bi 1 and other indicators,and plasma nf-kappaB activity,TGF-betal,SOD and GSH levels,analyzes the change and its relationship with type 2 diabetes peripheral neuropathy.At the end of the dosing 24 weeks,measure of serum GPT,GOT,BUN,Cr,analysis the bad influence of the body of long-term dosing drugs in mice.Results1.Quality standards research of the effective parts of Rhizoma anemarrhenae cortex phellodendriThrough the experimental investigation,extraction process parameters is Rhizoma anemarrhenae,cortex phellodendri medicinal material separate pieces,through the sieve no 3.Finally established the best extraction procedure,eight times the amount of 80%ethanol for the first time,soaking 0.5 h,80? reflux extraction 1.5h,the second six times the amount of 40%ethanol,80? reflux extraction 1.5h.Purification process research using PIPO-00 macroporous adsorption resin(the first column)with 732 cation exchange resin(the second column)in series,high pressure pump on a sample,sample liquid concentration on crude drug for 0.125g/mL,Separate elution,the first column in turn to 40%,70%ethanol elution velocity of 5-10ml/min elution,the second column using 0.5 mol/L ammonia water solution,mergerof eluent,vacuum concentration,vacuum drying,get the purity of the effective group is 50.0%or above.By 4 batches of pilotscale experiment,for the medicine suits of total saponins,total alkaloids and flavonoids effective parts of extraction,purification,concentration further experiment,Using berberine hydrochloride,phellodendrine,timosaponin BII,mangiferin as reference to establish a TLC identification method for altogether effective parts.Create a UV quantitative method to determine the content of total saponins,total alkaloid,flavonoids in altogether effective parts.Use HPLC and HPLC-ELSD to quantitative the berberine hydrochloride,phellodendrine,timosaponin BII,mangiferin in effective parts.Preliminary established quality standard for altogether effective parts.2.The research results of Pharmacodynamicsa)5 weeks later after Injection of STZ,compared with the normal group,the weight of the model group increase significantly,and FPG significantly increased(P<0.05),indicating that type II diabetes building success.b)Administration 8,16 and 24 weeks,FPG and random blood glucoselevel in mice:compared witn normal group,blood glucose or moael group mice significantly increase,have the meaning of meter(P<0.001 or P<0.01);Compared with model group,the total extract parker has better reduce FPG and significantly inhibitory effect of random blood glucose levels(P<0.01 or P<0.001),and a dose dependent,the effect is better than the single drug groups,with the increase of drug delivery time,especially the effect for FPG is obviously better than metformin and xuezhikang:(P<0.05).c)Administration 8,16 and 24 weeks,TC level in mice:In fasting and feeding conditions,compared with normal group,model group TC in mice and significantly increased,with statistical significance(P<0.001 or P<0.01):Compared with model group,the total extract has better reduce TC and obviously inhibitory effect of eating TC levels on an empty stomach.d)TG levels in mice after administrating 8,16 and 24 weeks:Compared with the normal group,model group TG mice and significantly increases,statistically significant(P<0.001 or P<0.01);Compared with model group,the total extract has better reduce fasting TG and significantly inhibitory effect of TG in feeding level.Total alkaloid obvious effects of low doses of TG on an empty stomach,and a high dose of total alkaloid has good inhibition in mice eating TG;When dosing of 8 and 16 weeks,the total extract of fasting and eating parker TG mice has obvious effect,a dose dependent,and give medicine at 24 weeks,know the total extract parker low dose effect is slightly better than the high dose group,but no statistical difference was found between the two.e)For 8,16and 24 weeks of model group compared with normal plasma NF-kB activity,TGF-?1 and SOD increased significantly,GSH levels decreased significantly,the difference was statistically significant(P<0.001 or P<0.01),while the total extract parker after continuous dosing,can significantly reduce the NF-kB activity in mice plasma,and TGF-?1 and SOD level(P<0.001 or P<0.01 or P<0.05),at the same time,the total extract parker can significant rise plasma GSH in diabetes in mice(P<0.01).Conclusiona)The purity of Rhizoma anemarrhenae and Cortex phellodendri effecti ve parts more than 50%,and quality standards have been established.b)Rhizoma anemarrhenae and cortex phellodendri total extracts on experimental diabetic mice of DPN has obvious protective effect.c)Rhizoma anemarrhenae and cortex phellodendri total extract inhibit the NF-kB activity and reduce the TGF-?1 and SOD levels at the same time.Index influence the expression of inflammatory factor is more common anemarrhena,cortex phellodendri total extract against one of the mechanism of action of diabetic peripheral neuropathy in mice.