| Objective:Based on the correlation analysis of ingredients,medicinal properties and medicinal effects,to study the mechanism of changes in the production of Gastrodia elata from Jianchang Bangjiang.Methods:Part 1.Research on composition changes.The elevated gastrodia ginger use“the healthy intestines stick”the law preparation,the extraction withdraws with 70%ethyl alcohol.Dissolves after the water,the different spot uses the benzine,the chloroform,the ethyl acetate,the normal butyl alcohol and so on the different rank solvent extraction;Layer chromatography and UPHLC-MS/MS methods are ued to compare di fferent parts and study the differences between the compone nts of each part.Part 2.Research on changes in drug prope rt ies.Selects the SD big mouse,divides into the blank group,the model group,the masculine control group dry ginger(group,)the fresh elevated gastrodia high,middle and low monitoring team,the ginger elevated gastrodia high,middle and low monitoring team stochastically,each group of 10.The cold syndrome model was replicated by freezing and gavage with cold and cold medicines.After 15 days,different doses of medicine were given for 7 days,and the rats were sacrificed.The rat heart,liver,spleen,lung,kidney and other organs were taken,and the organ index was calculated;Take liver tissue to prepare homogenate,and measure SDH,Na+-K+-ATP and plasma TG,T4 content according to the biochemical kit instructions;use HE staining method to check and record the changes in liver pathological slices.Part 3.Research on changes in pharmacodynamics.(1)The effect on the anticonvulsant effect of mice.Select clean mice and randomly divide them into blank group,model group,positive control group,natural gastrodia elata group,and ginger gastrodia elata group.The pathological model was replicated by intraperitoneal injection of Nicosame in mice,and the incubation period of convulsions and convulsion persistence of mice in each group were observed.Time,convulsion grade score,mortality and pathological section,etc.,to investigate the effect of Gastrodia elata on the anticonvulsant effect of mice before and after processing.(2)Effect on the analgesic effect of mice.Select Kunming clean mice and painful qualified mice,and randomly divide them into 7 groups:blank group,aspirin control group,gastrodia elata water phase group,n-butanol group,ethyl acetate group,chloroform group,petroleum ether group,etc.The change of pain area at each extraction site of Tianhe was determined by hot plate method.Another kunming species of clean-grade mice,randomly divided into seven groups(grouped together),the use of ace tic acid tors ion method to compare the analgesic effect and effect of the various extraction sites.Results:Part 1.Research on composition changes.The thin layer chromatograph result indicated that,chloroform-methyl alcohol(9:1)has the good separation effect to the ethyl acetate and the normal butyl alcohol;The benzine-ethyl acetate(9:1)is suitable for the benzine,the chloroform and the ethyl acetate,N-butanol phase and water are separated equally;low-polar petroleum ether and chloroform phase,Petroleum ether-acetone(10:1)is used as an expander,but petroleum ether-acetone(8:2)is more suitable for petroleum ether phase separation.For UPHLC-MS/MS analysis,from the point of view of chemical properties,three new characteristic peaks,including compounds 2,10,11,appeared after processing of Gastrodia elata,but characteristic peak 12 was only found in raw Gastrodia elata;for the peak area,compared with raw Gastrodia elata,The peak areas of compounds 3 and 9 decrease,but the peak areas of other compounds increase;for the extraction phase,compared with the ethyl acetate of Gastrodia elata,the peak area of compounds 6,9,13,17 in the ethyl acetate phase of Gastrodia elata increases Among them,compounds 7,11,and 12 are all new characteristic peaks in the solvent phase after the production of Gastrodia elata;the peak areas of the remaining compounds are reduced,and compound 8 is a new characteristic peak in the ethyl acetate phase of raw Gastrodia elata.Part 2.Research on changes in medicinal properties.(1)As for the organ change index,compared with the blank group,the rat organ index in the model group changed significantly(P<0.05),and the lung change index was the largest(P<0.01);compared with the model group,the heart index was divided by There was no significant difference in the middle dose of Gastrodia elata group(P>0.05),the other groups were statistically significant(P<0.05);the liver index was significantly different except the middle dose Gastrodia elata group and the low-dose Gastrodia group(P<0.05)In addition,the other groups did not change much(P>0.05);the spleen index was changed significantly in the Gastrodia ginger medium dose group(P<0.05),and the lung index was significantly different in each group(P<0.05);the kidney index was only in the gastrodia elata group.There was a significant difference in the low-dose group(P<0.05).(2)The biochemistry experiment aspect,the model group and the blank group compare,the triglyceride drops few(P>0.05);Compares with the model group,each group slightly has the increase,but the difference between the groups was not statistically significant(P>0.05).(3)For liver tissue homogenate,compared with the blank group,SDH in the model group was significantly increased(P<0.05),and Na+-K+-ATP decreased(P<0.05);in each group medium SDH activity sex low higher than mode type group(P<0.05),the day of anesthesia and low dose groups was not significant drop,not having statistically significant(P>0.05).However,the Na+-K+-ATPase activity of the middle dose group of Gastrodia elata increased(P<0.05).Although the other groups increased,they were not statistically significant(P<0.05).(4)HE dyeing result,compares with the blank group,the intrahepatic vesicle lesions in the model group were of different sizes,vacuoles or microvesicles;compared with the model group,the Gastrodia ginger high-dose group and the positive group and the high-dose Gastrodia The shape is better.Part 3.Research on changes in pharmacodynamics.(1)For anticonvulsant effect.Compared with the blank group,the seizure rate in the model group was 100%,and the seizure fatality rate was 75%;compared with the model group,the seizure rate was 100%in both the Gastrodia elata and Gastrodia elata group,which was greater than that of the positive control group(87.5%),The incubation period of seizures was statistically significant(P<0.05).Compared with the model group,the incubation period of seizures in the positive control group,gastrodia elata group and ginger gastrodia elata group was significantly prolonged,and the difference between the groups was statistically significant(P<0.01);Compared with the blank group,the model group had obvious convulsions(P<0.01);compared with the model group,the positive control group and the Gastrodia elata group had a greater degree of convulsion inhibition,and the difference between the groups was statistically significant(P<0.01).Although there was inhibition in the group,it was not statistically significant(P>0.05).(2)For analgesia.Compared with the blank group,the model group had significantly longer pain areas(P<0.01);compared with the model group,the chloroform group,ethyl acetate group,n-butanol group,and water phase group of Gastrodia ginger had longer effects on the pain area(P<0.05).Conclusion:1.From the perspective of composition changes.1.The separation system given priority in this study can better separate the different effective parts of Gastrodia ginger.Using the solvent method to study the different extraction parts of Gastrodia ginger,it is found that the chemical properties and content of Gastrodia after ginger production have changed.2.From the perspective of medicinal changes.By replicating the pathological model of cold syndrome in rats,observing the changes in rat organs and biologically active substances before and after the treatment of Gastrodia elata The changes in the medicinal properties of the drug are related,and there is a correlation with the dose.3.From the perspective of changes in efficacy.Ginger can improve the anticonvulsant and analgesic effects of Gastrodia on Nicotsame.At the same time,the analgesic effect of ethyl acetate phase in Gastrodia elata is stronger than that of other parts,which laid a research foundation for screening effective parts of Gastrodia elata.It can be seen that from the research on the composition,medicinal properties and medicinal effect changes,after the Gastrodia elata processed with ginger,it has an impact on the composition,medicinal properties and medicinal effects.Improved the energy metabolism of model rats with cold syndrome,enhanced the anticonvulsant and analgesic effects of Gastrodia elata The site of action.Since the enhancement of the drug effect after processing is like also to the change of the composition,the change of the composition in this study involves the chemical properties and content.Although the effective parts of ginger Gastrodia have been initially clarified,further clarification of its effect components still needs in-depth research.Based on the entire experimental content,this study can provide scientific data support for the enhancement of the efficacy of Gastrodia after processing,and also provide a research basis for expanding the source of new drugs.Objective:Based on the correlation analysis of ingredients,medicinal properties and medicinal effects,to study the mechanism of changes in the production of Gastrodia elata from Jianchang Bangjiang.Methods:Part 1.Research on composition changes.The elevated gastrodia ginger use“the healthy intestines stick”the law preparation,the extraction withdraws with 70%ethyl alcohol.Dissolves after the water,the different spot uses the benzine,the chloroform,the ethyl acetate,the normal butyl alcohol and so on the different rank solvent extraction;Layer chromatography and UPHLC-MS/MS methods are ued to compare di fferent parts and study the differences between the compone nts of each part.Part 2.Research on changes in drug prope rt ies.Selects the SD big mouse,divides into the blank group,the model group,the masculine control group dry ginger(group,)the fresh elevated gastrodia high,middle and low monitoring team,the ginger elevated gastrodia high,middle and low monitoring team stochastically,each group of 10.The cold syndrome model was replicated by freezing and gavage with cold and cold medicines.After 15 days,different doses of medicine were given for 7 days,and the rats were sacrificed.The rat heart,liver,spleen,lung,kidney and other organs were taken,and the organ index was calculated;Take liver tissue to prepare homogenate,and measure SDH,Na+-K+-ATP and plasma TG,T4 content according to the biochemical kit instructions;use HE staining method to check and record the changes in liver pathological slices.Part 3.Research on changes in pharmacodynamics.(1)The effect on the anticonvulsant effect of mice.Select clean mice and randomly divide them into blank group,model group,positive control group,natural gastrodia elata group,and ginger gastrodia elata group.The pathological model was replicated by intraperitoneal injection of Nicosame in mice,and the incubation period of convulsions and convulsion persistence of mice in each group were observed.Time,convulsion grade score,mortality and pathological section,etc.,to investigate the effect of Gastrodia elata on the anticonvulsant effect of mice before and after processing.(2)Effect on the analgesic effect of mice.Select Kunming clean mice and painful qualified mice,and randomly divide them into 7 groups:blank group,aspirin control group,gastrodia elata water phase group,n-butanol group,ethyl acetate group,chloroform group,petroleum ether group,etc.The change of pain area at each extraction site of Tianhe was determined by hot plate method.Another kunming species of clean-grade mice,randomly divided into seven groups(grouped together),the use of ace tic acid tors ion method to compare the analgesic effect and effect of the various extraction sites.Results:Part 1.Research on composition changes.The thin layer chromatograph result indicated that,chloroform-methyl alcohol(9:1)has the good separation effect to the ethyl acetate and the normal butyl alcohol;The benzine-ethyl acetate(9:1)is suitable for the benzine,the chloroform and the ethyl acetate,N-butanol phase and water are separated equally;low-polar petroleum ether and chloroform phase,Petroleum ether-acetone(10:1)is used as an expander,but petroleum ether-acetone(8:2)is more suitable for petroleum ether phase separation.For UPHLC-MS/MS analysis,from the point of view of chemical properties,three new characteristic peaks,including compounds 2,10,11,appeared after processing of Gastrodia elata,but characteristic peak 12 was only found in raw Gastrodia elata;for the peak area,compared with raw Gastrodia elata,The peak areas of compounds 3 and 9 decrease,but the peak areas of other compounds increase;for the extraction phase,compared with the ethyl acetate of Gastrodia elata,the peak area of compounds 6,9,13,17 in the ethyl acetate phase of Gastrodia elata increases Among them,compounds 7,11,and 12 are all new characteristic peaks in the solvent phase after the production of Gastrodia elata;the peak areas of the remaining compounds are reduced,and compound 8 is a new characteristic peak in the ethyl acetate phase of raw Gastrodia elata.Part 2.Research on changes in medicinal properties.(1)As for the organ change index,compared with the blank group,the rat organ index in the model group changed significantly(P<0.05),and the lung change index was the largest(P<0.01);compared with the model group,the heart index was divided by There was no significant difference in the middle dose of Gastrodia elata group(P>0.05),the other groups were statistically significant(P<0.05);the liver index was significantly different except the middle dose Gastrodia elata group and the low-dose Gastrodia group(P<0.05)In addition,the other groups did not change much(P>0.05);the spleen index was changed significantly in the Gastrodia ginger medium d05e group(P<0.05),and the lung index was significantly different in each group(P<0.05);the kidney index was only in the gastrodia elata group.There was a significant difference in the low-dose group(P<0.05).(2)The biochemistry experiment aspect,the model group and the blank group compare,the triglyceride drops few(P>0.05);Compares with the model group,each group slightly has the increase,b u t t h e difference between the groups was not statistically significant(P>0.05).(3)For liver tissue homogenate,Compares with the blank group,the model group’ s succinic acid dehydrogenase obviously elevates(P>0.05)the sodium potassium three adenylic acid glucoside reduces(P>0.05);Compares with the model group,each group’s succinic acid dehydrogenation enzyme activity drops(P>0.05).Although there was a decline in the low-dose Gastrodia elata group,it was not statistically significant(P>0.05).However,In the elevated gastrodia monitoring team sodium potassium three phosphoric acid adenosinase activeness increases(P<0.05).Although other groups have the increase,but non-statistics significance(P<0.05).(4)HE dyeing result,compares with the blank group,the intrahepatic vesicle lesions in the model group were of different sizes,vacuoles or microvesicles;compared with the model group,the Gastrodia ginger high-dose group and the positive group and the high-dose Gastrodia The shape is better.Part 3.Research on changes in pharmacodynamics.(1)For anticonvulsant effect.Compared with the blank group,the seizure rate in the model group was 100%,and the seizure fatality rate was 75%;compared with the model group,the seizure rate was 100%in both the Gastrodia elata and Gastrodia elata group,which was greater than that of the positive control group(87.5%),The incubation period of seizures was statistically significant(P<0.05).Compared with the model group,the incubation period of seizures in the positive control group,gastrodia elata group and ginger gastrodia elata group was significantly prolonged,and the difference between the groups was statistically significant(P<0.01);Compared with the blank group,the model group had obvious convulsions(P<0.01);compared with the model group,the positive control group and the Gastrodia elata group had a greater degree of convulsion inhibition,and the difference between the groups was statistically significant(P<0.01).Although there was inhibition in the group,it was not statistically significant(P>0.05).(2)For analgesia.Compared with the blank group,the model group had significantly longer pain areas(P<0.01);compared with the model group,the chloroform group,ethyl acetate group,n-butanol group,and water phase group of Gastrodia ginger had longer effects on the pain area(P<0.05).Conclusion:1.From the perspective of composition changes.The separation system given priority in this study can better separate the different effective parts of Gastrodia ginger.Using the solvent method to study the different extraction parts of Gastrodia ginger,it is found that the chemical properties and content of Gastrodia after ginger production have changed.2.From the perspective of medicinal changes.By replicating the pathological model of cold syndrome in rats,observing the changes in rat organs and biologically active substances before and after the treatment of Gastrodia elata The changes in the medicinal properties of the drug are related,and there is a correlation with the dose.3.From the perspective of changes in efficacy.Ginger can improve the anticonvulsant and analgesic effects of Gastrodia on Nicotsame.At the same time,the analgesic effect of ethyl acetate phase in Gastrodia elata is stronger than that of other parts,which laid a research foundation for screening effective parts of Gastrodia elata.It can be seen that from the research on the composition,medicinal properties and medicinal effect changes,after the Gastrodia elata processed with ginger,it has an impact on the composition,medicinal properties and medicinal effects.Improved the energy metabolism of model rats with cold syndrome,enhanced the anticonvulsant and analgesic effects of Gastrodia elata The site of action.Since the enhancement of the drug effect after processing is like also to the change of the composition,the change of the composition in this study involves the chemical properties and content.Although the effective parts of ginger Gastrodia have been initially clarified,further clarification of its effect components still needs in-depth research.Based on the entire experimental content,this study can provide scientific data support for the enhancement of the efficacy of Gastrodia after processing,and also provide a research basis for expanding the source of new drugs. |
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