Study The Anti-osteoporosis Effect Of The Oligosaccharides From Radix Morindae Officinalis

ObjectiveTo study the extraction and purification method of the oligosaccharides from Radix Morindae Officinalis,then to research its effects on the ovarian cause osteoporosis model in rats and clarify the mechanism.MethodsOligosaccharides were extracted from Radix Morindae Officinalis with a certain concentration of aqueous ethanol,purified with activated carbon and D-900 ion exchange resin,recrystallized with different proportion of two-phase solvent,to reach higher purity based on previous research.Fructose-strong hydrochloric acid chromogenic method and high performance liquid chromatography evaporative light scattering detector(HPLC-ELSD)method were used to monitor the extraction and purification effect.72 SD female rats were divided into nine groups:blank group(KB)and sham-operation group(JS),model group(MX),positive control group(YD),low dose group(DD),middle dose group(DZ),high dose group(DG)of Radix Morindae Officinalis oligosaccharides,bajijiasu group(BJ),extraction of Radix Morindae Officinalis group(BT).The other seven groups were bilateral-ovaries-removed(ovariectomized,OVX)except the KB and JS group,the fat near ovaries of the same size as the ovaries were removed in JS group.Eight weeks after the operation,KB group,JS group,MX group were given saline,while YD group was given alendronate 0.1 mg/100g(0.1mg per hundred gram body weight),DD,DZ and DG group were given corresponding dose of the oligosaccharides 10 mg/100g,40 mg/100g,100 mg/100g,BJ group give bajijiasu solution 5 mg/100g,BT group give water-extraction of Radix Morindae Officinalis 100 mg/100g,once a day.BMD of the rat right tibia was detected after administration for 8 weeks,so as related indexs and factors,such as concentration of urinary calcium,creatinine,hydroxyproline(HOP)and type I collagen peptide(NTX)N end and serum alkaline phosphatase(AKP),serum tartrate-resistant acid phosphatase(StrACP)activity,osteocalcin(BGP),collagen type I N-terminal propeptide(PINP),osteoprotegerin(OPG),nuclear factor kappa-B receptor activation factors predominate ligands(RANKL)concentration in serum.ResultsThe high purity of oligosaccharides from Radix Morindae Officinalis can get after extraction of 40%aqueous alcohol(V/V),purification with activated carbon and D-900 ion exchange resin in turn,of which the total content of sucrose,kestose,nystose,lF-Fructofuranosylnystose and bajijiasu was up to 50%and the total oligosaccharides shared more than 90%.Anti osteoporosis pharmacodynamics experiment results showed that the bone mineral density decreased in ovariectomized(OVX)rats,and the oligosaccharides from Radix Morindae Officinalis could relief this decline with dose-effect relationship;the high dose group could even keep bone mineral density of ovariectomized(OVX)rats by compared to the normal level.By compared to the model group,the concentration of Urine hydroxyproline(HOP)and serum osteoprotegerin(OPG)were increased,while the concentration of receptor activator of nuclear factor kappa-B ligand(RANKL)and serum procollagen I N-terminal propeptide(PINP)were reduced significantly of the high dose group(DG).The concentration of serum osteoprotegerin(OPG)was increased of the middle dose group(DZ),while the concentration of serum procollagen I N-terminal propeptide(PINP)were declined significantly.The concentration of urine type I collagen N-terminal peptide(NTX)increased and the concentration of serum osteocalcin(BGP)reduced of the low dose group(DD).ConclusionThe extraction and purification methods of the oligosaccharides from Radix Morindae Officinalis was stable and reliable.The anti-osteoporosis mechanism of the oligosaccharides from Radix Morindae Officinalis may be that it can improve the OPG/RANK/RANKL pathway,by increasing the concentration of serum osteoprotegerin(OPG),bonding with receptor activation of nuclear factor kappa-B ligands(RANKL)competitively,and reducing the concentration of serum RANKL at the same time,thus inhibiting the activity of RANK to RANKL,eventually suppressing the formation of osteoclast,delay the reduce of bone mass.