| ObjectiveWith the rapid economic development,China has become a big consumer of alcohol,and thus a series of alcoholic liver diseases are triggered,which has become a serious public health problem.Since the treatment of alcoholic liver disease remains limited,Chinese herbal medicine is considered an important alternative.The development of traditional Chinese medicine health-care food which can protect alcoholic liver injury has an important social significance.A best ratio of rhodiola sachalinensis and codonopsis pilsolula was determinedin accordance with a serum pharmacological experiment in an earlier stage.The optimumprocessing conditions of rhodiola sachalinensis and codonopsis pilsolula extract spray-drying were selected through an orthogonal test,and the active ingredients before and after thespray-drying and antioxidant loss rate were measured.Liquor is used to induce a mouse acute alcoholic liver injury model.The liver protection function and functional mechanism of the rhodiola sachalinensis and codonopsis pilsolula compound preparationat different doses are compared.Our study provides a theoretical basis for the development of the health-care function of the rhodiola sachalinensis and codonopsis pilsolula compound preparation.Method1.70%ethanol was used to conduct a mix extract of the rhodiola sachalinensis and codonopsis pilsolula.By an orthogonal test,the optimum conditions of the spray-drying were preferably selected.2.HPLC was used to determine the contents of salidroside and lobetyolin in the rhodiola sachalinensis and codonopsis pilsolula compound preparation after the spray drying and the loss rate was calculated.The antioxidant loss rate was calculated by DPPH radical scavenging ability.3.In vivo experiment:A total of 80 mice were randomly divided into a control group,a model group,a positive control group,a rhodiola sachalinensis group,a codonopsis pilsolula group,and low,medium and high doses rhodiola sachalinensis and codonopsis pilsolula compound preparation group with 10 mice in each group.Based on serum pharmacological experiment in an earlier stage and animal in vivo experiments,lg-60kg-1 of rhodiola sachalinensis(crude drug)and1.5g·60kg-1 of codonopsis pilsolula(crude drug)were determined as the best human daily recommended dose.The drug doses were 5 times,10 times and 20 times for the mouse low,medium and high dose groups,that is,the mediumdose of rhodiola rosea(crude drug)was 0.167g·kg-1 and the medium dose of codonopsis pilsolula(crude drug)was 0.25 g-kg-1.By calculating the drug extract yield of the rhodiola sachalinensis and codonopsis pilsolula,0.0895 g·kg-1,0.179 g·kg-1 and 0.358 g·kg-1 of the rhodiola sachalinensis and codonopsis pilsolula compound preparation(dry powder)were at low,medium and high doses.Tiopronin(50mg·kg-1)was selected as a positive drug.Mice in the blank group and model group were given a gavage of distilled water(0.1 mL·10g-1)and mice in the rest groups were given a gavageof a corresponding drug(0.1 mL·10g-1)once a day for a consecutive 5 days.After the last administration for 30min,in addition to the blank group was given the same amount of distilled water,the mice in the remaining group were given a gavageof a 56 degree Red Star Erguotou(16mL·kg-1)for a single time,and the mice fasted without water forbidden.After fasting for 12h,the mice were weighed,their eyeballs were picked to take blood and the supernatant was waited for measurement.According to kits,the serum levels of AST and ALT were measured.After the mice were sacrificed by cervical dislocation,the livers were quickly taken out,the liver weight was recorded and the liver index was calculated according to the formula:liver index=liver weight(mg)/mouse weight(g).An appropriate amount of liver tissue was weighed to prepare a liver tissue homogenates and the supernatantwas tested.According to the kits,the contents of TG,GSH,SOD and MDA in the liver homogenates were measured.Results1.The orthogonal test results showed that the optimumspray dryingprocessing conditions of the rhodiola sachalinensis and codonopsis pilsolula extract were as follows:an air inlet temperature at 160℃,a relative density of the extract at 1.09 andan accessory material addition amount at 50%.After the conduction of the spray drying,the dry powder yield was calculated at 28.95%and the water content was 4.02%.2.The active ingredient content determination results showed that before spray drying,the salidroside content was 3.400 ± 0.015 mg·g-1 and after spray drying,the salidroside content was 3.209±0.019 mg·g-1,suggesting a loss rate of 5.56%.Before spray drying,the lobetyolin content was 0.304 ± 0.0016 mg·g-1 andafter spray drying,the lobetyolin content was 0.288±0.0016mg·g-1,suggesting a loss rate of 5.26%.3.The DPPH radical scavenging results showed that before spray drying,the IC50 value was 0.681±0.061 mg·mL-1 and after spray drying,the IC50 value was 0.718 ± 0.022 mg·mL-1,suggesting an anti-oxidant loss rate of 5.43%.4.The HPLC methodological study investigation results showed that the investigated active ingredients were salidroside and lobetyolin.The precision RSD were 1.43%and 1.46%respectively;the stability RSD were 1.37%and 1.61%respectively;the repeatability RSD were 1.36%and 1.36%respectively;and the average recovery rate RSD were 1.79%and 1.77%respectively.5.The decomposed recipe results showed statistically significant differences ineach of the indexes when comparing other groups with the model group.The ALT levels can be significantly decreased in the compound preparation medium dose group compared with the codonopsis pilsolula group(P<0.01).The content of MDA can be significantly decreased in the compound preparation medium dose group compared with the rhodiola sachalinensis group(P<0.01).6.The optimumdose screening experiment results showed that there were significant differences in each of the indexes between the model group and the control group(all P<0.05).There was no significant difference in the AST levels between the compound preparation low dose group and the model group,while there were significant differences in the indexes between other drug administration group and the model group(all P<0.05).Compared with the compound preparation lowdose group,the SOD content was significantly increased in the compound preparation medium dose group(P<0.05).7.Liver index results showed that there were no significant differences among each group.Conclusions1.The optimumspray dryingprocessing conditions of the rhodiola sachalinensis and codonopsis pilsolula compound preparation were as follows:an air inlet temperature at 160℃,a relative density of the extract at 1.09 and an accessory material addition amount at 50%.2.The salidroside content in the rhodiola sachalinensis and codonopsis pilsolula compound preparation was 3.209±0.019 mg·g-1 and thelobetyolin content was 0.288±0.0016 mg-g-1.3.Rhodiola sachalinensis combined with codonopsis pilsolula have protective effects on acute alcoholic liver injury in mice.The best dosage is medium dose(0.186 g·kg-1). |
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