Studies On The Microwave Degradation Process Of Total Saponins In Different Parts Of Panax Notoginseng And Its Bioactivities

Panax notoginseng(Burk)F.H.Chen is a traditional Chinese herbal medicine with many pharmacological effects such as spreading stasis and hemostasis,dispersing swelling and pain,and it has great potential for the exploitation and utilization in medicine.Saponins are the main active ingredients,generally can be divided into common saponins and rare saponins.Mostly,the rare saponins are formed by common saponins to change the side chain structure or removing some of the glycosides.These saponins are extremely low in content.In recent years,as the discoveries of new bioactivities of rare saponins,how to obtain rare saponins fast and efficiently become the hotspot issues in researches of Panax notoginseng.High pressure microwave extraction can rapidly increase the temperature and increase the vibration of chemical bonds,so as to quickly obtain the rare saponins.Related to the microwave degradation study of American ginseng leaves and ginseng,has fully proved that microwave treatment can contribute to the degradation of ginsenosides transformation as well as the improvement of biological activity.However,there are no reports on the studies of microwave-assisted degradation of Panax notoginseng.Therefore,the microwave degradation technology was used to degrade total saponins in different parts of Panax notoginseng in this article.The research contents and results include the following aspects:1.The study on microwave degradation process of total saponins in different parts of Panax notoginseng.1.1 The identification of degradation products.The UHPLC-MS was used to analyze the retention time and mass spectrometry information of the chemical constituents in the microwave degradation products of total saponins in different parts of Panax notoginseng.According to the relevant literature data,6 newly produced components were identified from the microwave degradation products of Panax notoginseng leaves saponins.It was identified by spectrum analysis as notoginsenoside SFt3 and ginsenoside 20(S/R)-Rg3,Rk1,Rg5,20(S)-Rh2.8 components were identified from the microwave degradation products of Panax notoginseng saponins,namely ginsenoside 20(S/R)-Rh1,Rk3,Rh4,20(S/R)-Rg3,Rk1,Rg5.1.2 The study on the process optimization of degradation product formation.On the basis of single factor experiments,,the sequence of factors affecting on the conversion yield of the total saponins from different parts of Panax notoginseng that after optimized by orthogonal experiments and response surface methods was: microwave degradation temperature> microwave degradation power > microwave degradation time.The optimum conditions for microwave degradation of the total saponins from different parts of Panax notoginseng is microwave degradation power 500 W,microwave degradation temperature 150°C,and microwave degradation time 20 min.2.The study on the biological activity of the microwave degradation products of Panax notoginseng leaves saponins(MPNLS).2.1 The anticoagulant effect of MPNLS.Compared with the blank group,MPNLS can significantly prolonged the plasma prothrombin time at the concentration of 25 mg/m L,and have a anticoagulant activity(p<0.001).Ginsenosides Rk1,Rg5,20(S)-Rh2 and notoginsenoside SFt3 of MPNLS all show a anticoagulant effect.2.2 The cell inhibition in vitro tumor of MPNLS.MPNLS and its notoginsenoside SFt3 and ginsenoside 20(S/R)-Rg3,Rk1,Rg5,20(S)-Rh2 have a inhibitory effect on Human cervical cancer(He LA)cells in vitro with the concentration-dependent,especially ginsenoside 20(S)-Rh2,with a IC50 value of 8.232 ?mol/L.2.3 Anti-inflammatory experiment of acute inflammation in mice of MPNLS.MPNLS high(200mg/kg),medium(100mg/kg)and low(50mg/kg)doses can significantly reduced the auricle swelling of mice induced by xylene,showing a dose-dependent relationship,the high and medium doses experiments results are significant(p<0.01,p<0.05).By assaing the content of SOD enzyme and PGE2 in rat plasma,it was found that the activity of SOD enzyme is significantly increased and the release of inflammatory mediator PGE2 is significantly inhibited at high dose(p<0.01),thereby increasing the body's ability to scavenge oxygen free radicals and alleviating the symptoms associated with inflammation.Similarly,MPNLS can significantly inhibit the acute inflammatory reaction caused by carrageenan-induced foot swelling at high(200mg/kg),medium(100mg/kg)and low(50mg/kg)doses(p<0.01,p<0.05);and the inhibition rate of swelling at 4 hours is high to 68.51% at the dose of 200 mg/kg.It was found that the production of MDA(p<0.05,p<0.05)and NO(p<0.01,p<0.05)are significantly inhibited at high and medium doses,thereby inhibiting the level of liposome peroxidation in the body to achieve the purpose of anti-inflammatory and swelling.3.The study on the bioactivities of the microwave degradation products of Panax notoginseng saponins(MPNS).3.1 The hemostatic effect of MPNS.It was found that PNS and SPNS have significant hemostatic effects at low concentrations of 1 mg/m L and 3 mg/m L(p<0.001)by comparing the effects of the total saponins of Panax notoginseng(PNS),the total saponins of the steamed Panax notoginseng(SPNS)and MPNS on hemostatic and promoting blood circulation,and both of them have a significant promoting blood circulation effect when the concentration at 20 mg/m L(p<0.001).However,SPNS show a significant hemostatic efficacy at different concentrations(p<0.05,p<0.001).Ginsenoside R1 and Rd of PNS have a significant promoting blood circulation efficacy at concentrations of 1 mg/m L and 2 mg/m L(p<0.05,p<0.01),while Rb1 exhibits significant hemostasis at all concentrations(p<0.01).Ginsenosides Rk1 and Rg5 of SPNS and MPNS have a significant promoting blood circulation effect at concentrations of 1 mg/m L and 2 mg/m L(p<0.05,p<0.01),and ginsenosides Rh4 and 20(S/R)-Rg3 show a significant hemostatic efficacy(p<0.05,p<0.01).3.2 The invigorating blood effect of MPNS.The total WBC and HGB concentrations in the blood of mice with SPNS and MPNS high(800 mg/kg),medium(400 mg/kg)and low(200 mg/kg)doses are significantly higher than those in the model group(p<0.01,p<0.001),indicating that both can improve the symptoms of blood deficiency mice caused by cyclophosphamide and acetophenone hydrazide,and have a invigorating blood effect.At the same time,medium and high doses of SPNS can significantly reduce the increase of spleen index in blood deficiency mice(p<0.05).MPNS can significantly increase the thymus index of blood deficiency mice at high dose(p<0.05),the spleen index of the blood deficiency mice also decreases significantly(p<0.05).This show that SPNS and MPNS have a protective effect on mouse immune organs,but there are no significant differences in the thymus index and spleen index of mice in the PNS group with the model group.