| Objection:The main flavonoids,polysaccharides and microelements of Puerariae Lobatae Radix（PLR）and Puerariae Thomsonii Radix（PTR）before and after processing were determined by HPLC,phenol concentrated sulfuric acid and inductively coupled plasma;the pharmacodynamics differences of PLR and PTR before and after processing in the treatment of diarrhea,hypoglycemia and blood pressure were compared;the processing mechanism of PLR and PTR pieces was based on the intestinal flora The purpose of this study is to provide reference for the establishment of quality standards and the study of processing mechanism of Pueraria.Methods and results:Comparative study of components:PLR and PTR are both called Pueraria in ancient books and before the 2000 edition of Chinese Pharmacopoeia.However,in order to improve the accuracy of medicine,the 2005 edition of Chinese Pharmacopoeia divides Pueraria into two varieties,PLR and PTR,according to the source of medicinal materials.Both of them have the same property and flavor and functional indications,so that the use of PLR and PTR pieces is confused in clinical practice.So is its composition and clinical effect consistent?PLR and PTR decoction pieces have two specifications:thick piece and non-standard small cube.However,in order to facilitate the adjustment and use in clinic,the small cube is often used.However,the influence of different cutting technology on the chemical composition of PLR and PTR decoction pieces is not clear.In addition,the processed pieces of PLR and PTR mainly include raw products,simmer products and vinegar products.Different processing methods have different components of PLR and PTR and the difference in the effect of drug efficacy should be further studied.Therefore,the contents of flavonoids,polysaccharides and microelements in pueraria root,pueraria powder and pueraria varieties with different specifications and processing methods were compared.（1.1）Determination of flavonoids by HPLCThe content changes of five main flavonoids in Pueraria,Pueraria and processed products（simmer products,vinegar products）were determined by HPLC.The results showed that there was significant difference between PLR and PTR,and the content of flavonoids in PLR was significantly higher than that in PTR;the highest content of 3 T-hydroxypuerarin and 3T-methoxypuerarin in Pueraria was raw PLR pieces,the highest content of daidzin and daidzein was simmer PTR pieces,the highest content of Puerarin was vinegar PTR pieces（P<0.05）;the highest content of 3 T-hydroxypuerarin in PTR was simmer cubes（P<0.05）;The content of puerarin,3 T-methoxypuerarin and daidzin is the highest in the decoction of simmer PTR pieces,and the content of daidzein is the highest in the raw PTR cubes（P>0.05）.In addition,the content of flavonoids in the PLR pieces is mostly higher than that in the PLR cubes,among which,the content of 3T-hydroxypuerarin in the PLR pieces is 52.3%higher than that in the PLR cubes,the content of Puerarin is 38.3%higher,and the content of 3 T-methoxypueraria lobata is 38.3%higher,The content of Daidzin in the thick tablet was 35%lower.（1.2）the content of polysaccharide was determined by phenol concentrated sulfuric acid methodThe polysaccharide content of PLR and PTR before and after processing was determined by phenol concentrated sulfuric acid method.The results showed that the polysaccharide content of PLR and PTR processed products was different,and the polysaccharide content of PTR was significantly higher than that of PLR,among which,the polysaccharide content of raw PTR pieces was the highest,and that of vinegar PLR pieces was the lowest;in PTR group,compared with raw PTR cubes,the polysaccharide content of Pueraria after simmering and vinegar processing was higher 16.7%,compared with raw PTR pieces,the content of polysaccharide decreased by 40%and 25%respectively after simmering and vinegar processing.It can be seen that different cutting technology can affect the content of polysaccharide after processing.In the PLR group,compared with the raw PLR cubes,the content of polysaccharide after simmering increased by 8%and after vinegar processing decreased by 4%.Compared with the raw PLR pieces,the content of polysaccharide after simmering increased significantly and after vinegar processing The content of polysaccharide decreased by 50%.（1.3）determination of trace elements by inductively coupled plasma mass spectrometryThe contents of Ca,K,Cu,Fe,Mg,Mn,Zn in PLR,PTR and the processed products were determined by ICP-MS.The results showed that the contents of Mn,Fe were the highest in vinegar PLR pieces.The contents of Cu were the highest in vinegar PLR cubes,The contents of Zn were the highest in vinegar simmer PLR cubes,The contents of Mg and K were the highest in raw PTR cubes,The contents of Ca were the highest in vinegar PLR pieces.Mn、Ca、K、Zn、Mg in Products of PLR after simmering and vinegar has no significantly different（P>0.05）.The content of Cu decreased in PLR significantly after vinegar processed（P<0.05）and increased significantly（P<0.05）in PLR pieces after vinegar treatment.The content of Fe decreased after simmering,but increased after vinegar process.There was no significant difference in the content of trace elements between PTR simmering and vinegar treatment products.Compared with PTR cubes,the content of Fe in PTR pieces is higher,about double that in PTR cubes.（2）comparative study of efficacyModern pharmacological research has found that the antidiarrheal effect of pueraria root is significantly enhanced after simmering,and the content of puerarin,daidzein and daidzein,the main effective material base,is also significantly increased;the content of Puerarin after vinegar processing is significantly higher than that after simmering,but there is no research report on the effect of pueraria root processed with vinegar on diarrhea,and there is little literature on the treatment of diarrhea before and after processing of Pueraria;Puerarin,Pueraria polysaccharide,flavonoids and pueraria decoction can reduce the blood glucose of diabetic rats,and puerarin has a good therapeutic effect on hypertensive patients,but there is no study on the effect of raw PLR and raw PTR and vinegar processed products and simmer processed products on diabetic and hypertensive rats.The content of PLR and PTR in different specifications before and after processing was determined.The raw product specifications of PLR and PTR,simmer products and vinegar products were selected to study the difference of anti-diarrhea,hypoglycemic and anti-hypertensive effects.（2.1）study on the difference of anti-diarrhea effect of PLR and PTR before and after processingThe diarrhea model induced by senna leaf was used to determine the diarrhea index of rats within 6 hours after the decoction of PLR and PTR raw products,simmer products and vinegar products was given respectively.The diarrhea index was used to compare the effect of PLR and PTR raw products,simmer products and vinegar products in the treatment of diarrhea.The results showed that the diarrhea index was as follows:simmer PLR group>simmer PTR group>vinegar PTR group>raw PLR group>vinegar PLR group>raw PTR group,The experimental results showed that the antidiarrheal effect of simmer groups was significantly increased（P<0.05）compared with raw group and model group,and control group（Montmorillonite powder）has no significant difference compared with simmer groups（P>0.05）.（2.2）study on the difference of hypoglycemic effect of PLR and PTR before and after processingThe diabetic model induced by streptozotocin was used to measure the changes of blood glucose concentration in each stage of rats before and after administration.The blood glucose concentration changes were taken as the index to compare the differences of hypoglycemic effect of PLR and PTR before and after processing.The results showed that compared with the model group,the blood glucose concentration of the control group,raw PLR group,simmer PLR group,vinegar PLR group and vinegar PTR group decreased significantly after 6 days of administration（P<0.05）;after 11 days of administration,the blood glucose concentration in the control group and the simmer PLR group decreased significantly（P<0.01）,and the blood glucose concentration in the vinegar PLR group and the vinegar PTR group decreased significantly（P<0.05）,After 16 days,the blood glucose concentration of control group,raw PLR group,vinegar PTR group,simmer PLR group and vinegar PTR group decreased significantly（P<0.01）,while that of simmer PTR group decreased significantly（P<0.05）.（2.3）study on the difference of antihypertensive effect of PLR and PTR before and after processingThe hypertension model induced by N-nitro-L-arginine methyl ester was used to compare the changes of blood pressure of rats in different stages of administration,and the systolic blood pressure of rats was used as the index to compare the antihypertensive effect of PLR and PTR before and after processing;compared with the model group,in the first week of administration,except for the blank group,there was no significant difference in each administration group（P>0.05）,in the second week of administration,in each administration groups,the raw PLR group,simmer PLR group,vinegar PLR group and control group has significant difference（P>0.01）,In the third week of administration,there was a significant difference（P<0.01）of raw PLR group,simmer PLR group,vinegar PLR group and control group,and there was a significant difference（P<0.05）between raw PLR group and vinegar PLR group,There was significant difference of raw PTR group,simmer PTR group and vinegar PTR group（P<0.05）.there has no significant difference between each groups（P>0.05）.（3）study on the processing mechanism of PLR and PTR based on intestinal flora SPF Rats were randomly divided into 7 groups.In addition to the blank group,the other groups were given the same amount of decoction of PLR,PTR,simmer products and vinegar products respectively for 14 days.After the last Administration for 1 hour,the fresh feces of rats in each administration group and blank group were collected by finger abdominal massage,and the fecal DNA was extracted,PCR amplification was carried out,and DNA sequencing technology was used to determine the species richness and species difference of intestinal flora in the faeces of rats in different administration groups,and to compare the effects of PLR and PTR on intestinal flora before and after processing.High throughput data results show that:（3.1）Compared with the blank group,lactobacillus、unculturedbacteriumfMuribaculaceae decreased,the abundance of Blautia、Bacteroides、Holdemanella、Prevotella9 and[Eubacterium]coprostanoligenesgroup all increased。lactobacillus was higest in simmer PLR group,Roseburia was higest in raw PLR group,Blautia was higher in raw PLR group and vinegar PLR group;lactobacillus was higest in raw PTR,Roseburia was higest in vinegar PTR group.（3.2）Compared with the results of chemical composition,the content of daidzin and daidzein in the simmer PLR group is the highest,which is consistent with the highest content of Lactobacillus in the simmer PLR group of intestinal flora.Lactobacillus can produce lactic acid through metabolism in the intestinal tract,which can inhibit the growth and reproduction of harmful bacteria,and enhance and protect the intestinal viscosity It is speculated that daidzin and daidzein may be one of the reasons for the increase of Lactobacillus abundance in intestine;the highest content of Puerarin in vinegar PLR group is the same as that of blautia in intestinal flora,and blautia is a main butyric acid producing strain Acid can promote intestinal synthesis of mucin,repair intestinal mucosa,maintain intestinal micro balance,promote insulin response,and promote vascular expansion,directly participate in blood pressure reduction.It is speculated that puerarin may be one of the reasons for increasing blautia（Brucella）abundance in the intestinal tract.（3.3）Compared with the results of efficacy comparison,the anti diarrhea effect of simmer PLR is the best,and the highest abundance of Lactobacillus is in the same group of intestinal flora.It is speculated that the anti diarrhea effect of Radix Puerariae may be related to increasing the abundance of Lactobacillus in the intestinal tract,inhibiting the growth of pathogenic bacteria,maintaining the balance of intestinal microorganisms and protecting the intestinal mucosal barrier;The effect of vinegar PLR on blood glucose was the best,and blautia（Brucella）was the most abundant in the intestinal flora.It was speculated that the effect of Pueraria on blood glucose might be related to the increase of blautia（Brucella）in the intestinal tract,the increase of short chain fatty acid butyric acid in the intestinal tract,and the increase of insulin response.The effect of vinegar PLR on blood pressure was the best,and blautia in the intestinal flora was the best It is speculated that the effect of lowering blood pressure may be related to the increase of blautia abundance in intestine,increased receptor[G Protein coupled receptor（GPCR）and olfactory receptor（olfr78）]combine to promote the β oxidation process of intestinal mucosa cells,consume the oxygen in intestinal cavity,cause intestinal microenvironment,promote the growth of beneficial bacteria,inhibit the proliferation of potential pathogenic microorganisms in intestinal cavity,and finally alleviate the immune inflammatory response related to hypertension,reduce the target organ damage of hypertension,and reduce blood pressure.conclusion:（1）there are significant differences in chemical components between PLR and PTR,among which,the flavonoids in PLR are higher than those in PTR,and the polysaccharide content in PTR is higher than that in PLR;the processing methods will have different degrees of influence on the chemical components in PLR,and the processing methods have no significant influence on the components in PTR.（2）the pharmacodynamics of PLR and PTR are not identical.simmer PLR and simmerPTR have antidiarrheal effect,simmer PLR has the best antidiarrheal effect,but the antidiarrheal effect of raw and vinegar products is not obvious;PLR and its processed products have hypoglycemic effect,the best antidiarrheal effect belong to vinegar PLR;the hypoglycemic effect of raw Pueraria and its processed products is not obvious,and the hypoglycemic effect of raw PTR become better after being simmer and vinegar treated;The effect of vinegar PTR on reduce blood glucose was the best;PLR and PTR had a certain degree of blood pressure lowering effect,and PLR had a stronger effect on blood pressure lowering than PTR,and vinegar PLR had the best effect on blood pressure lowering;with the prolongation of administration time,the effect of blood pressure lowering of PTR and processed products had more effective.the effect of decrease blood pressure is not significant of raw PTR and its processed products.It is recommended to use the simmer PLR pieces to treat diarrhea in clinic,use vinegar PLR pieces to reduce blood sugar and blood pressure.（3）It is speculated that the antidiarrheal effect of simmer PLR may be related to the increase of daidzin and daidzein,which can promote the production of Lactobacillus in the intestine,promote the metabolism of lactic acid in the intestine,inhibit the growth and reproduction of harmful bacteria,enhance and protect the intestinal mucosal barrier.The hypoglycemic and hypotensive effects of vinegar PLR may be related to the increase of Puerarin Zn Fe Cu Content,which can promote the production of butyric acid in blautia（Brucella）in the intestine,thus promoting the response of insulin,increasing the response to insulin and the receptor[G Protein coupled receptor（GPCR）and olfactory receptor（olfr78）]combine to promote the growth of beneficial bacteria,inhibit the proliferation of potential pathogenic microorganisms in the intestinal cavity,and finally alleviate the immune inflammatory response related to hypertension and diabetes. |
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