Studies On The Extraction Technology, Purification And Immune Functions Of Soluble Polysaccharides From Radix Cynanchum Bungei Decne

It is widely recognized that Cynanchum Bungei Decne plays multipleroles in pharmacological and biological function as a traditional Chineseherbal medicine. This paper is performed on extraction technology,purification and immune functions of polysacchrides of Radix CynanchumBungei Decne. The main results are as follows:1 Extraction technology Studies indicated that temperature, time and solid-liquid ratio are themain influencing factors on the extraction percent of PRCB. In order toknow the extraction technology of soluble PRCB, single factor tests weredone to analyze the influence of each factor in solid-liquid ratio(1:15,1:20,1:25), temperature (80℃,90℃,100℃) , time (1 h,2 h,3 h) andnumber of times (1,2,3); Orthogonal experiment design methods (L933)were also applied in solid-liquid ratio, temperature and time. Fresh CB was desiccated after being crushed and extracted time afertime with ethanol. Powder of Radix Cynanchum Bungei Decne wasextracted for hours in hot water according to designed solid-liquid ratio,temperature, time. Then, starch was got rid of the extracted liquor by 4山东农业大学硕士学位论文enzyme method; the absorbency of PRCB was measured by the method ofphenol-vitril. Finally. The content of polysacchrides in sample wascalculated on the base of criterion curve and conversion expression. By single factor tests and orthogonal experiment, it is ensured thatthe optimum solid-liquid ratio is 1:20, temperature is 100℃ and theextraction time is 2 hours, the extraction time number is 1.2 Purification of polysacchrides After powder of Cynanchum Bungei was extracted, PRCB were gotby a series of steps, such as eliminating starch with enzyme's method,getting rid of protein with Sevag's method, isolating with 95% alcohol andwashing with 80% alcohol, and so on. PRCB1-PRCB3 were obtained afterdealing with by DEAE-cellulose column with eluent of water and NaClsolution, dialyzing, concentrating, and desiccating. They were white,yellow, or brown floc solids. The yields of three fractions were 32.2%,9.8%,13.9%, respectively. PRCB1a was got from PRCB1 fraction byseparation on Sephadex G-100 column with eluent of water. It is a whitefloc solid and its yield is 28%. PRCB1a was identified as a homogeneouscomponent by methods of Sephadex G-100 and UV Spectrum. By IRSpectrum and Thin-layer chromatograms, PRCB1a was composed ofglucose.3 Functions of immune adjustment Functions of PRCB and PRCB1a mainly inclede: enhancement on Tlymphocytes; adjustment on immune organs; effect on celiac macrophageability of engulfing; effect on the delayed type hypersensitivity ability;influnce on macrophage engulfing carbon granula.3.1 Enhancement on T lymphocytes Each dose group of PRCB and PRCB1a had notable difference(P<0.01) compared with control group. These results indicated that PRCBand PRCB1a could obviously enhance transformation of T lymphocytes 5白首乌可溶性多糖的提取工艺,分离纯化及免疫功能研究out of rabbit. In the meanwhill, this function related to dose. Effect ofPRCB50 mg/(kg·d)and PRCB1a 100 mg/(kg·d)was the best amongthe doses.3.2 Adjustment on immune organs Each dose group of PRCB and PRCB1a can enhance weigh ofthymus and spleen.There were significant difference (P<0.01), comparingeffect of PRCB 150 mg/(kg·d)and PRCB1a 50 mg/(kg·d)and 100 mg/(kg·d)with control group on thymus index of mice. On spleen index,effect of PRCB 150 mg/(kg·d)and 50 mg/(kg·d)and PRCB1a 150 mg/(kg·d)had notable difference(P<0.01) compared with control group.These results show that polysacchrides of Cynanchum Bungei Decne haveremarkable influnce on immune organs in mice and related to dose.3.3 Effect on celiac macrophage ability of engulfing Each dose group of PRCB and PRCB1a can notablely enhance celiacmacrophage ability of engulfing in mice. Phagocytic rate in mice reach themost notable level.On phagocytic inde...