Quantification of Total Phenols, Total Flavonoids, Total Anthocyanins and Evaluation of Antioxidant and Antiradical Activities of Detarium Senegalense Extracts from Chad

The aim of the present work is to assess the value of Detarium Senegalense by determining the content of total phenols, total flavonoids and total anthocyanins, and by evaluating the free radical scavenging activity of Detarium Senegalense extracts. For this purpose, sequential extraction using solvents of increasing polarity was essential. The various extracts obtained underwent phytochemical and biochemical analyses. Phytochemical screening revealed the presence of flavonoids, alkaloids, tannins, polyphenols, anthocyanins and steroids/terpenes. Quantitative analysis of total polyphenols, total flavonoids and total anthocyanins yielded the following results: total flavonoids (0.803 ± 0029 mg EQ/100g P for acetone extract of roots and 0.871 ± 0.401 mg EQ/100g P for methanol extract of leaves);total polyphenols (23.298 ± 12.68 mg EAG/100g P for acetone extract of roots and 24.69 ± 0.49 401 mg EAG/100g P for methanol extract of leaves);total monomeric anthocyanins (44.697 ± 0.939 mg EC3G/100g P and 16.699 ± 0.193 mg EC3G/100g P respectively for acetone and methanol extracts of stem bark). DPPH free radical scavenging activity was 1.674 ± 0.023 mg/mL for the acetone extract and 0.934 ± 0.24 mg/mL for the methanol extract of roots. .

Establishment of an in vitro Tissue Culture and Rapid Propagation System using Lonicera hypoglauca and Content Assessment of Total Flavonoids and Chlorogenic Acid

Lonicera hypoglauca is a traditional Chinese medicinal plant.In this study,the tender young leaves of L.hypoglauca were used for the first time as the explants to establish a rapid in vitro propagation and regeneration system.The results revealed that the optimal time for disinfection of the explants was 8 min and the optimal medium for callus induction was MS+2,4-D 4.0 mg·L^(-1)+sucrose 30 g·L^(-1),with an average callus induction rate of 86.67%.The optimal medium to induce differentiation of callus to bud was MS+6-BA 1.0 mg·L^(-1)+NAA 0.10 mg·L^(-1)+sucrose 30 g·L^(-1),with an average germination rate of 83.33%.The optimal medium to induce multiplication was MS+6-BA 1.5 mg·L^(-1)+NAA 0.05 mg·L^(-1)+sucrose 30 g·L^(-1),with a multiplication coefficient of 5.42.The optimal medium for root induction was 1/2 MS+NAA 0.15 mg·L^(-1)+activated carbon 0.3 g·L^(-1)+sucrose 15 g·L^(-1),with an average rooting rate of 91.11%.The survival rate of tissue-cultured seedlings in nutrient soil cultivation medium was as high as 100%.The total flavonoid content and chlorogenic acid content in the explant,callus tissue and regenerated plant were 1.83%,2.27%,1.33%and 2.77%,1.83%,1.74%respectively.This study provides novel insights into the rapid propagation and mass production of L.hypoglauca seedlings at an industrial scale and that it exhibits important application value and future prospects.

Optimization of Extraction Process for Total Flavonoids from Penthorum chinense Pursh and Comparison of Their Contents from Different Parts

[Objectives]This study was conducted to optimize the extraction process of total flavonoids from Penthorum chinense Pursh and compare their contents from different parts.[Methods]Single factor and orthogonal experiments were designed to optimize the extraction process of total flavonoids from P.chinense Pursh with the volume fraction of ethanol,the ratio of material to liquid,heating reflux extraction time and extraction times as factors,and the content of total flavonoids as the index.A verification test was carried out.The optimized extraction process was adopted to compare the contents of total flavonoids from different parts of P.chinense Pursh.[Results]The best extraction process was extracting the powder of P.chinense Pursh for 2.0 h with 20 times of 55%ethanol by reflux twice.Under this condition,the contents of total flavonoids were 3.63%,8.90%,11.28%,and 4.36%from stems,leaves,flowers and whole grass of P.chinense Pursh,respectively.[Conclusions]The process is reasonable,feasible and stable,and can effectively extract total flavonoids from P.chinense Pursh.The contents of total flavonoids from different parts of P.chinense Pursh were quite different,and the value was higher in the leaves and flowers,so the proportions of leaves and flowers should be paid attention to in the industrial processing of P.chinense Pursh.

Determination of total flavonoids content in buckwheat and inhibition of α-amylase activity

Diabetes is one of the most difficult chronic diseases to cure in the world,which seriously affects people’s health and quality of life.Flavonoids in buckwheat can regulate blood glucose levels by inhibitingα-amylase activity.Therefore,sweet buckwheat produced in Inner Mongolia was used as the research object,and buckwheat fl avonoids were extracted by ultrasonic-assisted extraction method.Total fl avonoids content was determined by ultraviolet-visible spectrophotometry.With acarbose as the positive control,the inhibition test ofα-amylase was carried out by DNS colorimetry to study the inhibition behavior of fl avonoids onα-amylase activity.The results showed that the extraction process of flavonoids was stable and reliable,and the established method for the determination of flavonoids was simple,accurate and reproducible.The total flavonoids content of buckwheat samples was 2.706 mg/g,buckwheat total fl avonoids extraction solution had an inhibitory eff ect onα-amylase,and its median inhibition concentration(IC_(50))was 38.53 mg/mL.The results of this experiment provide a technical reference for the development and utilization of fl avonoids in Inner Mongolia sweet buckwheat,and provide a theoretical reference for the development and application of flavonoid-rich hypoglycemic food.

Determination of Total Flavonoids in Acanthopanax senticosus(Rupr. et Maxim.) Harms by Enzymolysis and Its Technological Optimization

In order to optimize the ultrasonic extraction technique for the total flavonoid of leaf yellows plus, the contents of 21 leaf yellows plus total flavonoid from four regions in Heilongjiang Province were comparatively analyzed. The ultrasonic extraction technology was optimized by Box-Behnken response surface method, and the total flavonoid content of 21 kinds of Acanthopanax senticosus(Rupr. et Maxim.) Harms from different producing areas were analyzed by cluster analysis. The optimal process conditions were determined as ultrasonic time 30 min, solid-liquid ratio 1 : 12 and ultrasonic power 250 W, and the average yield of the total flavonoid was 1.453 mg·g^ (-1). By optimizing the ultrasonic-assisted extraction method, the total flavonoid content from different producing areas was compared in the experiment, which provided certain data support for the optimization of the extraction process in the future and laid a certain theoretical foundation for the quality analysis of Chinese medicinal materials.

Optimization of extraction process for total flavonoids of Sophorae Flos for the treatment of hyperlipidemia based on network pharmacology and molecular docking

This study aimed to investigate the mechanism of action of Sophora Flos(SF)in the treatment of hyperlipidemia(HLP)using network pharmacology and molecular docking methods,and to optimize the extraction process of the predicted active components.The STRING database was used for protein interaction analysis and PPI network construction via Cytoscape 3.9.1.Pymol was employed for docking and visualization.An extensive review of SF identifi ed 6 active ingredients,297 related objectives,84 disease objectives,and 57 total objectives.After protein interaction and topology analysis,18 core targets were identified.These included 146 gene function entries(P<0.05).Active compounds,mainly flavonoids,can modulate the expression of various proteins such as TNF,IL-6,IL-1β,PPARG,and TGFB1 to achieve therapeutic effects on HLP.The network pharmacology and molecular docking results suggested that the active fl avonoids component in SF may be related to the treatment of hyperlipidemia.Therefore,the orthogonal experiment method was used to optimize the extraction process of total fl avonoid from SF using ethanol refl ux extraction,based on a single factor experiment.The effects of refl ux time,solid-liquid ratio,ethanol concentration,and other factors on the extraction of total fl avonoid from SF were investigated.The optimum process conditions were refl ux time of 1.25 h,solid-liquid ratio of 1:15 g/mL and ethanol concentration of 60%.Using these conditions,the purity of total fl avonoid extracted from SF was 70.33±0.22%.

Determination of Total Flavonoids in Leaves of Tetracera asiatica

[Objectives] This study was conducted to determine and compare the contents of flavonoids in the leaves of Tetracera asiatica , an ethnic medicine from different habitats, in order to provide a basis for the quality control of its medicinal materials. [Methods] The flavonoid compounds in the leaves of T. asiatica were reflux-extracted in a water bath, and the total flavonoid content in T. asiatica was determined by ultraviolet-visible spectrophotometry. [Results] The total flavonoid content in T. asiatica leaves varied according to different habitats. The highest content was found in Gaofeng (19.41%) of Guangxi, followed by Huizhou (16.88%) of Guangdong and Hong Kong (16.76%). The lowest total flavonoid content was found in Shangcuntang (3.91%) of Bobai County, Guangxi, followed by Beiliu (4.15%), Guangxi. The total flavonoid contents in the samples ranged from 3.91% to 19.41%, with an average content of about 12.36%. [Conclusions] The UV spectrophotometry method is easy to operate, and has good repeatability, accurate detection results, and high reliability, and can be used for the determination of flavonoids in T. asiatica .

Effects of Ethanol Condensation Reflux Extraction and Ultrasound-assisted Extraction on the Content of Total Flavonoids of Pueraria edulis

With Pueraria edulis from different habitats in Yunnan Province,Guangdong Province and Guangxi Zhuang autonomous region as materials,the medicinal materials of P.edulis were extracted by ethanol condensation reflux extraction and ultrasound-assisted extraction,and the effects of different extraction methods the contents of total flavonoids in P.edulis from different areas were compared.The results showed that the best extraction time for P.edulis in Yunnan Province was 20 min,and that for Guangdong and Guangxi Zhuang Autonomous Region was 30 min.The yield of total flavonoids from P.edulis extracted by the condensation reflux extraction method was relatively better.The content of total flavonoids in P.edulis in Yunnan was the highest with an average content of 0.2557%,while the average content of total flavonoids in P.edulis in Guangdong was the lowest with an average content of 0.2108%.

Optimization of Extraction Conditions for Total Flavonoids from Fructus Aurantii Immaturus and Its Anti-UVB Radiation Activity

[Objectives]Optimum extraction conditions of total flavonoids from Fructus Aurantii Immaturus(TFFAI)and its resistance activity to ultraviolet radiation were investigated in present research.[Methods]The optimal extraction conditions of TFFAI were determined by single factor and orthogonal experiments,and the survival rate of TFFAI on HaCaT cells irradiated with UVB rays was investigated.It s antioxidant capacity was determined by ABTS.[Results]The results showed that the highest yield of TFFAI was obtained with 70%ethanol at a solid-to-liquid ratio of 1:50(w/v)and 40℃for 1.5 h by single-factor and orthogonal experiments.Total flavonoids(0.25-1.00 mg/ml)could significantly improve the survival rate of HaCaT cell line.Meanwhile,the maximum absorption peak of TFFAI was found at 283 nm,and in-vitro antioxidant experiment identified that TFFAI had a good clearance rate to ABTS.It suggestes that TFFAI could protect the cells from UVB damage by absorption of UVB rays and anti-oxidation.[Conclusions]TFFAI played a protective role on UVB irradiated cells through UVB ultraviolet absorption and antioxidant pathways.

Research Progress of Total Flavonoids from Plants in Trib.Lorantheae

Trib.Lorantheae is one of the important medicinal plants in traditional medicine.There are 41 genera,6 genera of which are produced in China,mainly distributed in Southwest China,South China and Central South China.There are many kinds of plants in this family,and 22 species can be used as Chinese medicinal materials in China.The branches and leaves of Trib.Lorantheae are rich in flavonoids,alkaloids,terpenoids,polysaccharides,organic acids and other functional substances,among which flavonoids are one of the important chemical components to exert pharmacological activity,and play an important role in hypoglycemic,lipid-lowering,anti-inflammatory,anti-tumor,anti-oxidation,anti-osteoporosis and so on.In this paper,the chemical composition,extraction method,component analysis and pharmacological action of flavonoids in Trib.Lorantheae plants were reviewed,in order to provide scientific reference for further development and clinical application of flavonoids in Trib.Lorantheae plants.

Optimization of Preparative Separation and Purification of Total Flavonoids from Radix Puerariae by Macroporous Resin Method

Aim To screen the optimum macroporous resin and conditions for the isolation and purification of flavonoids from Radix Puerariae. Methods The static and dynamic adsorption/desorption methods were used, and the separation and purification process was evaluated by measuring the concentration of total flavonoid in the fractions with UV spectrophotometer. Results The SP70 macroporous resin was the most effective compared with other macroporous resins. The optimum conditions were screened, which were 0.5 g· mL^- 1 corresponding to crude drug for concentration of extract, pH 5 - 6, and appended 60 times the volume of the resin bed （BV） with the adsorption speed 2 BV·h^-1, and the volume of aq. 70% （V/V） ethanol as eluant was 5 BV with desorption speed 2 BV·h^-1. By this method, the final contents of total flavonoids exceeded 80%. Conclusion The SP70 macroporous resin is the most effective one for large-scale isolation and purification of flavonoids from Radix Pueraria, which meets industrial needs.

Effect of total flavonoids of epimedium on liver microsomal CYP 1A2,CYP3A4 and CYP2E 1 activities in rats

To assess the potential effect of total flavonoids of epimedium （TFE） on cytochrome P450 and activity of its main isoforms in rat liver microsomes. TFE （300 mg/kg） was administered once daily to male Sprague-Dowley rats by gavage for fifteen days. The total cytochrome P450 content and its main isoforms CYP1A2, CYP3A4 and CYP2E1 activities in rat liver microsomes were detected. The activity of CYP1A2 was measured by fluorometry and the activities of CYP3A4 and CYP2E1 were determined by measuring the amount of methanal and p-aminophenol formed using UV/Vis spectrophotometer, respectively. Administration of TFE significantly increased the total CYP450 content and activities ofCYP1A2, CYP3A4 and CYP2E1 in rat liver microsomes, compared with the control group. Particularly, the activities of CYP1A2 and CYP2E1 were enhanced significantly （P〈0.01）. TFE induced the increase in total CYP450 content and its main isoforms CYP1A2, CYP3A4 and CYP2E1 activities in rat liver microsomes.

Total flavonoids of hawthorn leaves promote motor function recovery via inhibition of apoptosis after spinal cord injury

Flavonoids have been reported to have therapeutic potential for spinal cord injury.Hawthorn leaves have abundant content and species of total flavonoids,and studies of the effects of the total flavonoids of hawthorn leaves on spinal cord injury have not been published in or outside China.Therefore,Sprague-Dawley rats were used to establish a spinal cord injury model by Allen's method.Rats were intraperitoneally injected with 0.2 m L of different concentrations of total flavonoids of hawthorn leaves(5,10,and 20 mg/kg)after spinal cord injury.Injections were administered once every 6 hours,three times a day,for 14 days.After treatment with various concentrations of total flavonoids of hawthorn leaves,the Basso,Beattie,and Bresnahan scores and histological staining indicated decreases in the lesion cavity and number of apoptotic cells of the injured spinal cord tissue;the morphological arrangement of the myelin sheath and nerve cells tended to be regular;and the Nissl bodies in neurons increased.The Basso,Beattie,and Bresnahan scores of treated spinal cord injury rats were increased.Western blot assays showed that the expression levels of pro-apoptotic Bax and cleaved caspase-3 were decreased,but the expression level of the anti-apoptotic Bcl-2 protein was increased.The improvement of the above physiological indicators showed a dose-dependent relationship with the concentration of total flavonoids of hawthorn leaves.The above findings confirm that total flavonoids of hawthorn leaves can reduce apoptosis and exert neuroprotective effects to promote the recovery of the motor function of rats with spinal cord injury.This study was approved by the Ethics Committee of the Guangxi Medical University of China(approval No.201810042)in October 2018.

The Optimization of Ultrasonic Extraction Technology of Total Flavonoids in Leaves of Mallotus apelta by Response Surface Analysis Methodology

[Objectives] Response surface analysis methodology was used to optimize the extraction technology of total flavonoids from Mallotus apelta leaves. [Methods] The total flavonoids were extracted using ultrasonic assisted ethanol extraction,and extraction rate of total flavonoids was taken as evaluation index. On the basis of single-factor experiment,Box-Behnken response surface methodology was employed for selecting the optimum extraction process. [Results]The optimum extraction conditions of total flavonoids were as follows: 75% of ethanol concentration,1∶ 25 of ratio of material to liquid,31 min of ultrasonic time. Under these conditions,the extraction rate of total flavonoids was 1. 115%.[Conclusions] The extraction process obtained by response surface methodology was stable,reasonable,accurate and reliable. It was a feasible method to extract the total flavonoids from M. apelta leaves.

Effects of Total Flavonoids ofHippophae RhamnoidesL.on Intracellular Free Calciumin Cultured Vascular Smooth Muscle Cells of Spontaneously Hypertensive Rats and Wistar-Kyoto Rats

To explore the effects of total flavonoids of Hippophae rhamnoides L. （TFH） quercetin （Que） and isorhamnetin （Isor） on the intracellular free calcium （[Ca^2＋]） in vascular smooth muscle cells （VSMC） of spontaneously hypertensive rats （SHR） and Wistar-Kyoto rats （WKY）. Metheds： Fluo 3-acetoxymethylester（Fluo-3/AM） was used to observe the effects of TFH （100mg/L） and its essential monomers, namely Que （10^-4mol/L） and Isor （10^-4mol/L） on changes of [Ca^2＋]1 in cultured SHR and WKY VSMC （abbr. to Ca-SHR ＆ Ca-WKY） following exposure to high K^＋, norepinephrine （NE） and angiotensin Ⅱ （AngⅡ）, and to compare with the effects of verapamil （Ver）. Results： （1） TFH, Que and Isor had inhibitory effects on resting Ca-SHR （P〈0.05）, but had no significant effects on Ca-WKY （P〉0.05）. （2） High K^＋ could increase Ca-SHR more significantly than Ca-WKY （P〈0.05）; TFH, Que and Isor could inhibit the elevation of [Ca^2＋]1 induced by high K^＋ -depolarization, with the effects similar to that of Ver, and the effect on Ca-SHR was more significant than that on Ca-WKY （P〈0.05）. （3） NE and Ang Ⅱ could increase Ca-SHR more significantly than Ca-WKY （P〈0.05）, TFH, Que and Isor had remarkably inhibitory effect on the elevation of Ca-SHR and Ca-WKY induced by NE or Ang Ⅱ. （4） In the absence of extracellular Ca^2＋ , TFH, Que and Isor also had certain inhibitory effect on Ca-SHR and Ca-WKY induced by NE, and the effect on the former was more significant than that on the latter（P〈0.05）. Ceaclusiea： TFH, Que and Isor might decrease the levels of [Ca^2＋], in VSMCs by blocking both voltage-dependent calcium channels （VDC） and receptoroperated calcium channels （ROC） in physiological or pathological state, which may be one of the important mechanisms of their hypotensive and protective effects on target organs in patients with hypertension.

Radix Ilicis Pubescentis total flavonoids combined with mobilization of bone marrow stem cells to protect against cerebral ischemia/reperfusion injury

Previous studies have shown that Radix Ilicis Pubescentis total flavonoids have a neuroprotective effect, but it remains unclear whether Radix Ilicis Pubescentis total flavonoids have a synergistic effect with the recombinant human granulocyte colony stimulating factor-mobilized bone marrow stem cell transplantation on cerebral ischemia/reperfusion injury. Rat ischemia models were administered 0.3, 0.15 and 0.075 g/kg Radix Ilicis Pubescentis total flavonoids from 3 days before modeling to 2 days after injury. Results showed that Radix Ilicis Pubescentis total flavonoids could reduce pathological injury in rats with cerebral ischemia/reperfusion injury. The number of Nissl bodies increased, Bax protein expression decreased, Bcl-2 protein expression increased and the number of CD34-positive cells increased. Therefore, Radix Ilicis Pubescentis total flavonoids can improve the bone marrow stem cell mobilization effect, enhance the anti-apoptotic ability of nerve cells, and have a neuroprotective effect on cerebral ischemia/reperfusion injury in rats.

Determination of Total Flavonoids in Leaves of Paliurus ramosissimus and Orthogonal Experiment Optimization Process

[Objectives] To screen the optimal extraction conditions of flavonoids from leaves of Paliurus ramosissimus( Lour.) Poir.,measure the content of flavonoids in leaves of P. ramosissimus. using the Microplate Reader,and provide experimental basis for the development of medicinal resources of P. ramosissimus. [Methods] The ultrasonic extraction method was adopted and orthogonal design was carried out to study the effects of ethanol concentration( A),ultrasonic time( B),the solid-to-liquid ratio( C) on the extraction of flavonoids from leaves of P. ramosissimus. The Microplate Reader was used to measure the content of flavonoids in leaves of P. ramosissimus. in different extraction conditions,P. ramosissimus. and the optimal extraction conditions were used to measure the content of flavonoids from leaves of P. ramosissimus.[Results]Effects of the ultrasonic extraction method on the extraction rate of flavonoids from leaves of P. ramosissimus: extraction times >ethanol concentration > Ultrasound time > solid-liquid ratio. The screened optimal extraction process was: ethanol concentration of 65%,the solid-to-liquid ratio of 1∶ 25,ultrasonic time of 2 h and extraction times Under this condition,the extraction rate of total flavonoids in leaves of P. ramosissimus. was 2. 56%. [Conclusions] This method is simple and feasibility,reliable,and suitable for rapid measurement with large samples.

Study on Ultrasonic Extraction of Total Flavonoids from Angelica keiskei Koidzumi

[Objective] This study aimed to optimize the ultrasonic extraction process of total flavonoids from Angelica keiskei Koidzumi. [Method] Using the yield of total flavonoids as an indicator, the ultrasonic extraction process of total flavonoids from A. keiskei was optimized by orthogonal experimental design. [ Result ] Among four factors in orthogonal experimental design, extraction time exhibited the most significant effect on the extraction result; ethanol concentration exhib- ited a significant effect on the extraction result; ultrasonic power and solid-to-liquid ratio had no significant effect. The optimal conditions for ultrasonic extraction of total flavonoids from A. keiskei were extraction time of 25 min, ethanol concentration of 80%, ultrasonic power of 60 W, solid-to-liquid ratio of 1：20 （g/ml）, under which the yield of total flavonoids reached 1.56%. [ Conclusion] This study provides the basis for further development and utilization of total flavonoids from A. keiskei.

Optimization of Extraction Process of Clerodendrum philippinum Schauer var. simplex Mlodenke Total Flavonoids(CPTF) by Central Composite Design-Response Surface Methodology

[Objectives] The research aimed to optimize extraction process of Clerodendrum philippinum Schauer var. simplex Mlodenke total flavonoids( CPTF),and provide reference for its development and utilization. [Methods] Based on single-factor test,ethanol concentration,extraction temperature and extraction time were taken as independent variables,and total flavonoids yield was taken as dependent variable. The test was conducted according to central composite design principle. Multivariate linear regression and binomial equation fitting of the result were conducted,and extraction process of CPTF was optimized by using response surface methodology. [Results]The optimal extraction process of CPTF was as below: ethanol concentration 54. 76%,extraction temperature 83. 92℃,extraction time 102. 64 min,solid-liquid ratio 1:20,extraction for twice. [Conclusions] The extraction process of CPTF by central composite design-response surface methodology was simple and feasible,with reliable prediction result,which was suitable for industrial production.

Inflammatory Mechanism of Total Flavonoids of Chrysanthemum and Medicated Serum on Castrated Dry Eye Animal and Cell Models

Objective To observe the effects of total flavonoids of chrysanthemum and medicated serum on the expression of related proteins in the lacrimal tissue and dry-eye cell models of male rabbits with dry eye caused by castration.Methods(1)150 male Japanese rabbits were randomly divided into five groups,with 30 rabbits in each group:normal control group(group A),sham group(group B),model group(group C),androgen control group(group D)and total flavonoids of chrysanthemum treatment group(group E).The androgen deficiency dry-eye model was established by bilateral castration in groups C,D and E.Normal saline was administered to groups A,B and C by gavage;androgen(testosterone propionate)was injected into muscle in group D;and group E was given total flavonoids of chrysanthemum by gavage.All white rabbits were tested the Schirmer I test(SIT)and tear break-up time(BUT).After euthanasia,tear gland tissue was harvested so that we could observe pathological changes in the expression of related inflammatory factors in the lacrimal gland tissue.The expression of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and transforming growth factor-β1(TGF-β1)was detected in the lacrimal gland tissue by immunohistochemistry.Reverse transcription PCR was used to quantitatively detect expression of TGF-β1 mRNA.(2)Male Wistar rat lacrimal epithelial cells were used to establish a model of eye stem cell apoptosis caused by androgen levels.The blank control group was set up without androgen culture,the control group with androgen culture,and the total flavonoids of chrysanthemum group without androgen.The MTT method was used to determine the optimal intervention dosage of drug-containing plasma.Western blot and QPCR were used to detect the expression of AR mRNA,NF-κB phosphorylated protein and TGF-β1 in lacrimal epithelial cells,and the androgen-like effect of total flavonoids of chrysanthemum was observed.Results(1)Immunohistochemistry showed that groups A,B,D and E had significantly lower expression of IL-1βand TNF-αthan group C(P<0.05);among these,group E had slightly higher expression than group D(P>0.05).RT-PCR results showed that the relative expression of TGF-β1 mRNA in groups A,B,D and E was significantly higher than in group C(P<0.05),and the relative expression of TGF-β1 mRNA in groups D and E was higher than that in groups A and B(P<0.05).(2)Using the MTT method,the final concentration of interfering cells was calculated to be 13.2%.The expression of AR protein,NF-κB and TGF-β1 in the chrysanthemum flavonoid plasma intervention and testosterone propionate intervention groups was enhanced,and there were significant differences relative to the blank group(P<0.01).The expression level of NF-κB in the total flavonoid containing plasma intervention group was lower than that in the testosterone propionate intervention group(P<0.01).Conclusions The total flavonoids of chrysanthemum can inhibit IL-1βand TNF-αexpression in the lacrimal gland tissue of castrated male rabbits with dry eye to increase synthesis of TGF-β1 mRNA and TGF-β1,thereby inhibiting the inflammatory response.The medicated plasma with total flavonoids of chrysanthemum promotes expression of AR mRNA,upregulating expression of NF-κB,further promoting upregulation of TGF-β1 protein expression in lacrimal epithelial cells,inhibiting inflammation by regulating related proteins,and ultimately alleviating the symptoms of dry eye.