MicroRNA expression profile and lipid metabolism characteristics in liver of rat undergoing high-fat diet

This study aimed to investigate the microRNA expression profile and the characteristics of lipid metabolism in the livers of rats undergoing a high-fat diet.Fifty male Sprague-Dawley(SD)rats were divided into a standard chow group(C group,N=10)and a high-fat diet group(H group,N=40).After 12 weeks,the rat body weight,body length,fat mass,and serum lipid concentration were measured.The expression profile of microRNAs and the gene and protein expression levels involved in lipid metabolism in rat liver were detected.Body fat and serum lipid concentrations were all significantly higher in the H group than those in the C group(p<0.05 or p<0.01).The expression of 10 microRNAs showed significant differences in the liver(p<0.05).In particular,the let-7 family expression levels significantly increased(p<0.05)in the H group compared with those in the C group.Compared with the C group,the high-fat diet resulted in low FAS,CPT1A,and ApoAI mRNA expression levels(p<0.05 or p<0.01)and high PPARαand FAT/CD36 mRNA expression levels in the H group rat liver(p<0.01).Meanwhile,the protein PPARα,FAS,CPT1A,FAT/CD36,and ApoAI expression levels were all significantly lower in the H group than those in the C group(p<0.05 or p<0.01).In conclusion,the high-fat diet increased the body fat and serum lipid levels and altered the 10 microRNA expression levels in the liver.The high-fat diet may affect hepatic carbohydrate metabolism and increase ectopic fat accumulation through let-7 family overexpression.The high-fat diet for 12 weeks decreased lipid metabolism level in the liver,thereby decreasing fatty acid synthesis,oxidation,and transport by down-regulating the PPARα,FAS,CPT1A,FAT/CD36,and ApoAI protein levels.

Chiral metabolism of propafenone in rat hepatic microsomes treated with two inducers

AIM: To study the influence of inducers of drug metabolism enzyme, beta-naphthoflavone (BNF) and dexamethasone (DEX), on the stereoselective metabolism of propafenone in the rat hepatic microsomes. METHODS: Phase I metabolism of propafenone was studied using the microsomes induced by BNF and DEX and the non-induced microsome was used as the control. The enzymatic kinetics parameters of propafenone enantiomers were calculated by regress analysis of Eadie-Hofstee Plots. Propafenone enantiomer concentrations were assayed by a chiral HPLC. RESULTS: The metabolite of propafenone, N-desalkylpropafenone, was found after incubation of propafenone with the rat hepatic microsomes induced by BNF and DEX. In these two groups, the stereoselectivity favoring R(-) isomer was observed in metabolism at low substrate concentrations of racemic propafenone, but lost the stereoselectivity at high substrate concentrations. However, in control group, no stereoselectivity was observed. The enzyme kinetic parameters were: (1) K(m). Control group: R(-) 83+/-6, S(+) 94+/-7; BNF group: R(-) 105+/-6, S(+)128+/-14; DEX group: R(-) 86+/-11, S(+) 118+/-16; (2)V(max). Control group: R(-) 0.75+/-0.16, S(+) 0.72+/-0.07; BNF group: R(-) 1.04+/-0.15, S(+)1.07+/-14; DEX group: R(-) 0.93+/-0.06, S(+) 1.04+/-0.09; (3)Cl(int). Control group: R(-) 8.9+/-1.1, S(+) 7.6+/-0.7; BNF group: R(-) 9.9+/-0.9, S(+)8.3+/-0.7; DEX group: R(-) 10.9+/-0.8, S(+) 8.9+/-0.9. The enantiomeric differences in K(m) and Cl(int) were both significant, but not in V(max), in BNF and DEX group. Whereas enantiomeric differences in three parameters were all insignificant in control group. Furthermore, K(m) and V(max) were both significantly less than those in BNF or DEX group. In the rat liver microsome induced by DEX, nimodipine (NDP) decreased the stereoselectivity in propafenone metabolism at low substrate concentration. The inhibition of NDP on the metabolism of propafenone was stereoselective with R(-)-isomer being impaired more than S(+)-isomer. The inhibition constant (Ki) of S(+)- and R(-)-propafenone, calculated from Dixon plots, was 15.4 and 8.6 mg x L(-1), respectively. CONCLUSION: CYP1A subfamily(induced by BNF) and CYP3A4 (induced by DEX) have pronounced contribution to propafenone N-desalkylation which exhibited stereoselectivity depending on substrate concentration. The molecular base for this phenomenon is the stereoselectivity in affinity of substrate to the enzyme activity centers instead of at the catalyzing sites.

Effect of vitamin A and E on carbohydrate and lipid metabolism in diet-induced obese wistar rats

In the present study, we investigated the biochemical alterations and gene expression of carbohydrate and lipid metabolism after oral administration of vitamin A and E for 2 months in diet-induced obese Wistar rats. Vitamin A and E administration reduced significantly the increase in body weight and food intake and normalized the alterations in lipid profiles in obese rats compared to normal rats. Moreover, both vitamins decreased the fat accumulation in liver tissues of obese rats. Finally, they up-regulated mRNA expression of Pyruvate Kinase (PK) and Glucose Transporter-2 (GLUT-2), and increased lipolysis and cholesterol metabolism through up-regulation of lipoprotein lipase (LPL), Sterol Responsible Element Binding Protein-1a (STREBP-1a) and STREBP-1c mRNA expression. In conclusion, vitamin A and E regulate gene expression of carbohydrate and lipid metabolism, and also ameliorate changes associated with obesity induced by high fat diet in Wistar rats.

<i>In Vitro</i>Competitive Metabolism Study of Olmesartan Medoxomil in Rat Liver S9 Fractions Using LC/MS

Olmesartan Medoxomil (OLM), Ramipril (RPL) & Fenofibric acid (FA) are used to treat hypertension and cardiovascular disease. These drugs undergo hydrolytic metabolism by the enzyme liver esterase and converts into their respective active metabolites Olmesartan (OL), Ramiprilat (RPT) and Fenofibric acid (FA) for OLM, RPL and FEN respectively. In this study the competitive metabolism of OLM, in presence of RPL and FEN was investigated in rat liver s9 fractions using a validated LC-MS method. Olmesartan Medoxomil was found to be highly reactive to the rat liver S9 fractions and formation of active metabolite Olmesartan is highest. The rate of formation of active metabolite Olmesartan reduced by 12.68% in the presence Ramipril and 6.56% in presence of Fenofibrate. A marked reduction of 18.96% was found in the formation of active metabolite Olmesartan from Olmesartan Medoxomil when all the three drugs are in combination.

Identification of the metabolite and cytochrome P450 isoforms involved in rat liver microsomal metabolism of TM208

To identify the metabolite and CYP450 isoforms involved in rat liver microsomal metabolism of TM208. The present study investigated the metabolism of TM208 and the effects of selective CYP450 inhibitors on the metabolism of TM208 in rat liver microsomes. Various specific inhibitors of CYP were used to identify the isoforms of CYP involved in the metabolism of TM208. The inhibitor of CYP2D and that of CYP2B had strong inhibitory effects on TM208 metabolism in a concentration-de- pendant manner, the inhibitor of CYP1A had a modest inhibitory effect, and the inhibitor of CYP3A seemed not to have an obvious inhibitory effect on TM208 metabolism. TM208 might mainly be metabolized by CYP2D and CYP2B in rat liver microsomes.

Metabolism of Mequindox in Isolated Rat Liver Cells

Mequindox （MEQ）, 3-methyl-2-quinoxalinacetyl-l,4-dioxide, is widely used in Chinese veterinary medicine as an antimicrobial agent and feed additive. Its toxicity has been reported to be closely related to its metabolism. To understand the pathways underlying MEQ＇s metabolism more clearly, we studied its metabolism in isolated rat liver cells by using liquid chromatography coupled with electrospray ionization hybrid linear trap quadrupole orbitrap （LC-LTQ-Orbitrap） mass spectrometry. The structures of MEQ metabolites and their product ions were readily and reliably characterized on the basis of accurate MS2 spectra and known structure of MEQ. Eleven metabolites were detected in isolated rat liver cells, two of which were detected for the first time in vitro. The major metabolic pathways reported previously for in vitro metabolism of MEQ in rat microsomes were confirmed in this study, including N O group reduction, carbonyl reduction, and methyl monohydroxylation. In addition, we fotmd that acetyl hydroxylation was an important pathway of MEQ metabolism. The results also demonstrate that cellular systems more closely simulate in vivo conditions than do other in vitro systems such as microsomes. Taken together, these data contribute to our understanding of the in vivo metabolism of MEQ.

Effect of Different Calcium Supplements on Bone Metabolism in Rats

Osteoporosis, characterized by loss of bone mass and microarchitectural deterioration of bone tissue, results in enhanced bone fragility and increases risk of fractureIll. In China, the incidence of primary osteoporosis is as high as 50%-70% in 60-69 years old females and approximately 30% in 60-69 years old males[21, which is closely related with the low intake of calcium. According to the nationwide nutrition and health survey in 2002 in China, the average daily calcium intake of Chinese residents is 391 mg, accounting for 41% of the recommended calcium intake.

Effect of Starvation-Refeeding Status on Cholesterol Metabolism in Rats Fed High-Cholesterol Diet

The present study investigated the effect of starvation-refeeding status on cholesterol metabolism in rats fed a high-cholesterol diet or a cholesterol-free diet. Twenty male and 20 female Donryu rats (age 5 weeks) were fed a cho-lesterol-free diet for 14 days. Then the male and female rats were each divided into two groups: feeding and starva-tion-refeeding groups. The feeding groups were fed the experimental diet for 3 days, and the starvation-refeeding groups fasted for 2 days followed by 3 days of feeding. Half of each of groups was fed a cholesterol-free diet and the other half was fed a high-cholesterol diet. Starvation-refeeding significantly increased the plasma free cholesterol and HDL-cholesterol concentrations in both the high-cholesterol-diet-fed rats and the cholesterol-free-diet-fed rats. In the female rats, plasma total cholesterol and cholesteryl ester concentrations were significantly higher in the high-cholesterol groups than in the cholesterol-free groups, whereas TG concentration and total cholesterol/TG ratio were not significantly different among all of the groups. Liver total cholesterol and cholesteryl ester were significantly higher in the high-cholesterol groups than in the cholesterol-free groups in both male and female rats. These results suggest that starvation-refeeding affected cholesterol metabolism at least in part. The reactivity of the cholesterol me-tabolism may be different between male and female rats.

Oxidative Metabolism of Estrone Modified by Genistein and Bisphenol A in Rat Liver Microsomes

Genistein, the main isoflavone from soy, and bisphenol A （BPA）, a food contaminant, are considered ubiquitous xenoestrogens. Here we investigated the influence of genistein and BPA on estrone （El） metabolism in rat liver microsomes. Both substances inhibited the 2-hydroxylation and 16a-hydroxylation of E1, but in different degrees, thereby reducing the 2-OH-E1/16a-OH-E1 ratio,

In situ absorption and metabolism of stilbene glycoside in rat

Stilbene glycoside(TSG)has been shown to have many beneficial properties.It is therefore essential to understand the absorption and metabolism of TSG in detail.We determined the recovery of TSG and its metabolites(TSG sulfate/glucuronides)in rat gastric contents,gastric mucosa,portal vein plasma,celiac arterial plasma,bile,and urine after administration of 15 mg of TSG in 0.5 mL physiological saline or incubation for 20 min in situ in the stomach of rats.Within 20 min,(64.0±9.8)% of the administered TSG disappeared from the stomach;later,TSG was recovered in both free and conjugated forms in plasma and bile,but not in urine.On the other hand,only free TSG was detected in the gastric contents and mucosa;it was also detected in the portal vein plasma as(48.1±3.5)% of the total TSG(all forms of TSG).However,the proportion of free TSG in the celiac arterial plasma and bile decreased to 4%-10%.In addition,the proportion of free TSG to total TSG in the liver microsome incubation mixture after TSG was incubated in liver microsome at 37 ℃ for 30 min was very low [(10.6 ± 2.6)%].These results indicate that TSG could be quickly absorbed from the rat stomach,conjugated in liver and excreted in bile.Such novel information would be helpful for the use of TSG as a beneficial natural product which may improve its proposed efficacy in preventing chronic diseases.

Changes of Selenium Metabolism Glutathione Peroxidase Activity and Lipid Peroxides Content in Severely Scalding-injured Rats

The changes of sclenium metabolism, glutathione peroxidase activity and lipid peroxidescontent in the tissues of rats suffering from 30% TBSA full thickness scalding were observed in thefirst 7 days after injury. It was found that selenium content in the rat tissues decreased remarkably af-ter injury, which in turn resulted in serious reduction of glutathione peroxidasc activity and significantincrease of lipid peroxides in the scrum, crythrocytcs and liver. However the muscular tissue showedno significant changes. These facts imply that after burn injury, the body is in a state of selenium deficiency, the lossof selenium might be responsible for the reduction of anti - peroxidation ability of glutathioneperoxidase, and conscqucntly there is an increase of lipid peroxides in the tissues. Only the musculartissue is insensitive to lipid peroxidation. It is believed that the reduction of anti-peroxidation abilityof glutathione peroxidasc after bum injury might be one of the main causes to intensify, the injury re-suiting from free radicals.

Anomeric specificity of D-glucose metabolism study in rat and human erythrocytes

The anomeric specificity of D-glucose metabolism in erythrocytes has been since 1985 the matter of extensive investigations reported in about ten publications. The present report aims at providing an integrated review of the major findings on this issue.

Nutritional Metabolism of Antarctic Krill Product Protein in Rats

This work conducted a four-week metabolism test on rats to study the digestion and absorption characteristics of five protein-based krill products prepared from Antarctic krill as raw material.It aimed to provide theoretical support for the effective use of Antarctic krill protein and the development of novel protein resources.The results showed that the weight gain and true digestibility of the rats fed with krill meat,surimi and ordinary krill powder were significantly higher(P<0.05)than those of the rats fed with traditional casein.Compared to casein,proteins from the five Antarctic krill products were found to significantly improve the net protein utilization(P<0.05),and reduce the total cholesterol and triglycerides in the serum of rats(P<0.05).In summary,the Antarctic krill protein-based products with high nutritional values can be used as a potential novel protein resource in the food industry.

Disposition and Metabolism of 1 -Nitropropane in Rats and Chimpanzees

The metabolic fate of 1 -nitropropane (1-NP) has not been previously reported. In this study male rats and chimpanzees were given single doses of 40 mg/kg ip and 5 mg/kg iv 1-[1-^(14)C]NP, respectively. The quantitative extent of urinary and fecal elimination was similar in both species. The rats excreted 16.5% ofthedosein urine and 1.7% in feces. For chimpanzees the respective values were 14.8 and 1.2%. Experiments with rats demonstrated that the major route of elimination was by exhalation. With a total elimination via the lungs of 72.6%, rats expired 10.3% of the dose as unchanged 1-NP. Five polar metabolites were isolated from the urine of chimpanzees. The two major metabolites were identified as 3-hydroxypropionic acid and 7V-methyl-N-2-(methylsulfinyl)ethylpropionic acid amide (NMPA). Both substances were also excreted in rat urine. The two identified metabolites indicate that 1-NP was degraded to propionic acid, part of which was modified to 3-hydroxypropionic acid or NMPA. A hypothetical pathway for the biochemical generation of NMPA is suggested.(c)1989 Academic Press, Inc.

Anti-hypertensive effects of rosiglitazone on renovascular hypertensive rats:role of oxidative stress and lipid metabolism

This study investigated the anti-hypertensive mechanismof rosiglitazone in renovascular hypertensive rats,and examined its relationship to oxidative stress and lipid metabolism. The renovascular hypertension was induced by stenosis of the left renal artery. Four groups of rats were selected： control,induced untreated,rosiglitazone（ 20 mg / kg） and captopril（ 10 mg / kg）. After 14 d of administration,compared with induced untreated group,rosiglitazone group reduced the renovascular hypertensive rats ＇ systolic blood pressure and diastolic blood pressure,and decreased total cholesterol（TCH）,triglyceride（TG）,angiotensin II（ Ang II） and angiotensin receptor（ AT1） levels（ P ＆lt; 0. 05）. Meanwhile,rosiglitazone remarkably decreased the levels of malondialdehyde（ MDA） and hydrogen peroxide（ H2O2） while improved the levels of supperoxide dismutase（ SOD） and reduced glutathione（ GSH）. These results suggested that rosiglitazone could effectively decreased the blood pressure in renovascular hypertensive rats,and this might be performed by regulating the activity of angiotensin and the lipid metabolismand improving the oxidative stress.

Effects of Lujingyiqishengxue Pill on Iron Metabolism in Rats with Iron Deficiency Anemia

[Objectives]This study was conducted to observe the effect of Lujingyiqishengxue Pill on iron metabolism in rats with iron deficiency anemia. [Methods] The iron-deficiency anemia rat model was established by feeding low-iron diet. Meanwhile, the rats were given oral gavage of ferrous succinate(0.036 g/kg, positive drug group) and Lujingyiqishengxue Pill(4.4, 2.2, 1.1 g/kg, high, middle and low dose groups), once daily for 42 consecutive days. The body weight of the rats was observed every week, and the peripheral blood[red blood cells(RBC), hemoglobin(HGB), and hematocrit(HCT)]and the iron contents in tissues(the liver, spleen, small intestine, kidney) of the rats were detected after modeling;and serum iron(SI), serum total iron binding capacity(TIBC), transferrin saturation(TSAT), serum ferritin(SF) and serum transferrin receptor 1(TFR1) and other iron metabolism indexes were determined. [Results] Compared with the model group, the high-dose Lujingyiqishengxue Pill significantly reversed the peripheral blood(HGB, HCT) and iron contents of various tissues(the liver, spleen, small intestine, kidney) in rats(P<0.01), and significantly increased SI, TSAT, SF(P<0.01), while the contents of TIBC and TFR1 were significantly decreased(P<0.01). [Conclusions] Lujingyiqishengxue Pill can significantly improve anemia and regulate iron metabolism in rats with iron-deficiency anemia, which provides a pharmacological reference for the clinical application of Lujingyiqishengxue Pill.

Influence of rotating magnetic field on cerebral infarction volume, cerebral edema and free radicals metabolism after cerebral ischemia/reperfusion injury in rats

BACKGROUND: It has shown that magnetic field can improve blood circulation, decrease blood viscosity, inhibit free radicals, affect Ca2+ flow in nerve cells, control inflammatory and immunological reaction, and accelerate nerve cell regeneration. In addition, protective effect of magnetic field, which acts as an iatrophysics, on ischemic brain tissues has been understood gradually. OBJECTIVE: To investigate the effects of rotating magnetic field (RMF) on volume of cerebral infarction, cerebral edema and metabolism of free radicals in rats after cerebral ischemia/reperfusion injury. DESIGN: Randomized controlled animal study. SETTING: Rehabilitation Center of disabled children, Liaoniang; Department of Rehabilitation, the Second Affiliated Hospital, China Medical University; Department of Rehabilitation Physiotherapy, the First Affiliated Hospital, China Medical University. MATERIALS: A total of 70 healthy Wistar rats aged 18-20 weeks of both genders were selected and randomly divided into 3 groups: sham operation group with 12 rats, control group with 20 rats and treatment group with 38 rats. The treatment group included 4 time points: immediate reperfusion with 6 ones, 6-hour reperfusion with 20 ones, 12-hour reperfusion with 6 ones and 18-hour reperfusion with 6 rats. Main instruments were detailed as follows: magnetic head of rotating magnetic device was 6 cm in diameter; magnetic induction intensity at the surface of magnetic head was 0.25 T in silence; the maximal magnetic induction intensity was 0.09 T at the phase of rotation; the average rotating speed was 2500 r per minute. METHODS: The experiment was carried out in the China Medical University in March 2003. Focal cerebral ischemic animal models were established with modified Longa’s method. Operation was the same in the sham operation, but the thread was inserted as 10 mm. Neurologic impairment was assessed with 5-rating method to screen out cases. Those survivals with grade 1 and grade 2 after ischemia for 2 hours and reperfusion for 24 hours were included in the control group and treatment group. Those in the sham operation group and control group were not treated with RMF. Magnetic head was directed towards the head of rats of the treatment group, and the magnetic head was about 7 mm from skin, treated for 15 minutes. The rats were decapitated to take out brains at 24 hours after reperfusion in each group. Water content of brain and volume of cerebral infarction were assessed with wet-dry weight method and TTC staining, respectively. Activity of superoxide dismutase (SOD), content of malondialdehyde (MDA) and change of brain histomorphology in brain tissue of ischemic side were analyzed. MAIN OUTCOME MEASURES: ① Volume of cerebral infarction and changes of water content in brain; ② measurements of SOD and MDA contents in brain tissue of rats in all groups. RESULTS: A total of 70 qualified animals were involved in the final analysis after rejecting the death and unqualified animal models. ① Water content of brain: Water content of brain in the treatment was less than that in the control group at any time point except the immediate time point, and cerebral edema was relieved [(2.48±0.22)%, (2.32±0.19)%, (2.23±0.36)%, (2.91±0.44)%, P < 0.05]. In addition, there were no significant differences among 6-hour, 12-hour and 18-hour reperfusion groups (P > 0.05). ② Volume of cerebral infarction: The absolute volume of cerebral infarction in the treatment group was smaller than that in the control group [(128.21±15.05), (171.22±40.50) mm3, t =2.438, P < 0.05], and the relative volume of cerebral infarction was smaller than that in the control group [(20.22±1.44)%, (25.17±3.85)%, t =2.95, P < 0.05]. ③ Contents of SOD and MDA in brain tissues: Compared with the control group, the SOD content in the brain tissue in the treatment group increased [(54.54±3.85), (69.52±5.88) kNU/g, t =5.568, P < 0.05], while the MDA content decreased [(0.85±0.06), (1.03±0.09) μmol/g, t =4.076, P < 0.05]. ④ General morphological observation: General morphology manifested that the edema was distinct in the right cerebral hemisphere in the control group, showing fat-like white, shallow anfractuosity, flat gyria, brittle tissue and easy to break up. The edema of right cerebral hemisphere was light and surface was hyperaemia in the treatment group. CONCLUSION: RMF may improve anti-oxidative ability of brain tissue of rats with acute focal cerebral ischemia/reperfusion injury and reduce volume of cerebral infarction and degrees of cerebral edema.

Rat Hepatocyte Glucose Metabolism Is Affected by Caloric Restriction but not by Litter Size Reduction

This work aimed at investigating whether the liver glucose metabolism could have a role in any change of glucose homeostasis that might exist in the reduced-litter rat at the age of 60 days. Additionally, post-weaning caloric restriction and its hepatic effects were explored, as this intervention is reported as a useful measure against obesity and its related disturbances. The animals were raised in litters of nine （control, CG） or three pups during lactation. These small-litter pups were FG （fed freely） or subjected to 30% RG （caloric restriction） after weaning until 60 days of age. The increased adiposity induced by lactational overfeeding was not reversed by caloric restriction. Hepatocyte glucose metabolism and glucose tolerance test were not affected by litter size, but caloric restriction increased liver basal glucose release, diminished gluconeogenesis and retarded the glycemic decay during the insulin tolerance test. Liver glucose metabolism of young adult rats was not affected by lactational overfeeding. Up to this age, however, moderate caloric restriction had a potent influence that might compromise whole-body glucose homeostasis and prompt to insulin resistance.

Long-Term Animal Feeding Trial of the Refined Konjac Meal Ⅰ. Effects of the Refined Konjac Meal on the Calcium and Phosphorus Metabolism and the Bone in Rat

Effects of refined konjac meal (RKM) on the calcium and phosphorus metabolism and bone parameters were observed in rats of both sexes fed with food containing 1% of RKM for 18 months. A comparable group of rats fed on basic diet only was used as a control.Results obtained indicate that all the measured parameters (serum calcium and phosphorus level, femur weight and its calcium and phesphorus content, and the osteometry of the tibia) showed no significant difference between the experimental and the control groups.Thus there is no adverse effect either on the calcium and phosphorus metabolism or on the bone after a long-term intake at a moderate dosage.

The bio-active components of the Mongolian medicine Horcha-6 and therapeutic mechanism in the rat migraine model

Background:The active components of Horcha-6 were identified using liquid chromatography with tandem mass spectrometry.Also,we investigated the potential mechanisms that explain why Horcha-6 may be effective in treating migraines through the use of network pharmacology and a rat migraine model.Methods:After identifying the active components of Horcha-6,the corresponding genes of the active components’target were obtained from the Universal Protein database,and a“compound-target-disease”network was constructed using Cytoscape 3.9.0 software.For the in vivo experiments,nitroglycerin was injected intraperitoneally into rats to create a migraine model.Pre-treatment with Horcha-6 was administered orally for 14 days,and rats were subjected to migraine-related behavior tests.RNA sequencing was performed to identify the gene expression regulated by Horcha-6 in the trigeminal nerve.Results:A total of 903 chemical components of Horcha-6 have been collected in the liquid chromatography with tandem mass spectrometry.We discovered 55 of the Horcha-6 bio-active components that were evaluated based on their Percent Human Oral Absorption(≥30%)and DL values(≥0.185)on the traditional Chinese medicine systems pharmacology database.The“compound-target-disease”network contained 163 intersection targets with the migraine state.Gene Ontology analysis indicated that these components significantly regulated the immune response,vascular function,oxidative stress,etc.When Kyoto Encyclopedia of Genes and Genomes enrichment analysis was performed,we observed that most of the target genes were significantly enriched in the inflammation and neuro-related signaling pathway,toll-like receptor signaling pathway,neuroactive ligand-receptor interaction,etc.These predictions were further demonstrated via in vivo animal model experiments.The RNA sequencing results showed that 41 genes were down-regulated(P<0.05)and 86 genes were up-regulated(P<0.05)in the Horcha-6 treated group compared with the untreated group.Those genes were mainly involved in neuromodulation,vascular function,and hormone metabolism.Conclusion:The 55 bio-active components in Horcha-6 regulate inflammation,hormone metabolism,and neurotransmitters and have potential as a therapy to treat migraines.