Anethole Research Articles

Trans-anethole ameliorates LPS-induced inflammation via suppression of TLR4/NF-κB pathway in IEC-6 cells

This study was undertaken to investigate the protective role of trans-anethole (TA) in lipopolysaccharide (LPS)-induced rat intestinal epithelial cells (IEC-6) injury and the potential mechanisms. The cells were pretreated with TA (0 and 1 mM) for 24 h, prior to stimulation by LPS (1 mg/mL) for 24 h. Compared with the control group (CON), LPS stimulus resulted in decreased cell viability, intestinal barrier injury, increased cell apoptosis and cell cycle arrest at the G2/M phase. These effects triggered by LPS were reversed by TA. In order to reveal the main genes and pathways involved among the groups, transcriptome analysis was performed to identify the differential expression genes (DEGs) among the treatment groups. There were a total of 493 DEGs (275 upregulated and 218 downregulated) that were identified between the LPS and CON group. Meanwhile, a total of 361 DEGs (103 regulated and 258 downregulated) were identified in the LPS+TA group compared with the LPS group. The results showed that the DEGs were mostly enriched in immune related pathways, such as tumor necrosis factor (TNF) signaling pathway, cytokine-cytokine receptor interaction, complement and coagulation cascades, interleukin-17 (IL-17) signaling pathway, NF-kappa B (NF-κB) signaling pathway, antigen processing and presentation, and NOD-like receptor signaling pathway. Based on the results of RNA-sequencing, further investigation of the signaling pathway involved revealed that TA could inhibit the activation of toll like receptor 4 (TLR4)/NF-κB signaling pathway and NLR family pyrin domain containing 3 (NLRP3) inflammasome in LPS-induced IEC-6 cells. In conclusion, this finding demonstrated a functional role of TA in intestinal epithelial cells injury and indicated that TA may be a potential strategy for treatment of inflammatory intestinal diseases.

Anticonvulsant Activity of trans-Anethole in Mice.

Epilepsy is a chronic neurological disorder affecting 1-2% of world population, and one-third of patients are refractory to pharmacological treatment. This fact has stimulated research for new antiepileptic drugs and natural products have been an important source. trans-Anethole (TAN) is a phenylpropanoid, component of some essential oils, extracted from plants, and its effects have been little studied. Therefore, this study is aimed at investigating the TAN effect in classic seizure models and evaluate the electroencephalographic (EEG) profile of animals treated with this substance. For this, Swiss male mice (Mus musculus) were used, and the lethal dose was evaluated and subsequently submitted to the test maximal electroshock (MES), the pentylenetetrazole- (PTZ) induced seizure test, and the EEG profile. Initially, the LD50 for TAN was estimated in 1000 mg/kg (i.p.) dose and there was no sign of acute toxicity or death. In the MES test, TAN 300, i.p. (12.00 ± 2.9 s) and 400 mg/kg, i.p. (9.00 ± 4.4 s) doses was able to decrease tonic seizures duration induced by electric discharge (0.5 mA, 150 pulses/s, for 0.5 s). In the PTZ test (75 mg/kg, i.p.), TAN 400 mg/kg, i.p. increased the latency to myoclonic jerks (80.0 (56.0–134.0)), the latency totonic-clonic seizures (900.0 (861.0–900.0) and decrease seizure duration (0.0 (0.0–10.0)). No deaths were found in this groups compared to vehicle. EEG analysis showed an amplitude decrease of waves (ratio of baseline) in TAN 300 (1.82 ± 0.23) and 400 mg/kg (1.06 ± 0.16) groups. In this way, TAN at 400 mg/kg was able to inhibit and/or attenuate seizures by increasing the time for the onset of spasms and convulsions, as reducing the duration of seizures. The EEG profile corroborate with this results showing a reduction in the amplitude of waves compared to the PTZ group. Thus, TAN showed an anticonvulsant effect in all experimental models performed, behavioral and electroencephalographic.

Trans-anethole ameliorates lipopolysaccharide-induced acute liver inflammation in broilers via inhibiting NF-κB signaling pathway

The aim of this study was to investigate the protective effect of trans-anethole (TA) on lipopolysaccharide-induced acute liver inflammation model of chickens by determining the levels of inflammatory mediators in serum and liver, relative mRNA expression and protein expression of inflammation-related genes in NF-κB signaling pathway. A total of 160 one-day-old male chickens (Arbor Acres) were assigned into 4 treatments with 8 replicates of 5 birds each. On d 20, the control group was intraperitoneally injected with sterile saline and the other groups were injected with lipopolysaccharide (LPS; 5 mg/kg body weight). There were no significant differences in average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) among groups. However, compared with the control group, the LPS group significantly increased (P < 0.01) the serum levels of interleukin-6 (IL-6), interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α), alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and decreased (P < 0.01) the interleukin-10 (IL-10) level. TA attenuated (P < 0.01) these increases in IL-1β, TNF-α, ALT, and AST levels and improved (P < 0.01) the IL-10 level. In liver, the groups fed with TA had lower (P < 0.01) concentrations of IL-6 and TNF-α as well as higher (P < 0.05) concentration of IL-10. Furthermore, TA downregulated (P < 0.05) the mRNA expression levels of nuclear factor kappa B p65 (NF-κB p65) and TNF-α, also upregulated (P < 0.05) IL-10 and inhibitor of NF-κB alpha (IκBα) upon LPS challenge. In protein level, supplementation of 600 mg/kg of TA downregulated (P < 0.05) and upregulated (P < 0.05) the protein expression of NF-κB p65 and IκBα, respectively. The present findings suggest that TA could alleviate the acute liver inflammation induced by LPS via blocking the activation of NF-κB and the 600 mg/kg of TA plays more fruitful role in protecting broilers against LPS stimulus.

Transanethole protects against Global Cerebral Ischemia through its Anti-inflammatory and Anti apoptotic activity

Aim and Objective: In this research study, transanethole at a dose of 250 and 500mg/kg p.o was investigated for its potency against Bilateral Common Carotid Artery occlusion (BCCAO) for 1 hr accompanied by 22 hrs reperfusion. Method: Healthy Albino Wistar rats (200–250gm) were divided randomly into 4 groups (n=9). Group I was considered as sham control, received 2% tween 80p.o, group II was considered as ischemic- reperfusion (disease control) and received 2% tween 80p.o and group III and IV were considered as prophylactic treatment groups and received transanethole at doses of 250mg/kg, p.o and 500mg/kg, p.o. respectively. After pretreatment with transanethole for14 days, rats were subjected to bilateral common carotid artery occlusion (BCCAO) for 1 hour accompanied by 22 hr reperfusion (I/R). After 22 hrs of reperfusion, motor coordination, behavioral assessment, infarct area, brain water content, excitatory mediators, inflammatory and apoptotic markers were determined. Results: Transanethole improved the motor coordination, lowered the brain water content, infarction volume and attenuated the levels of excitatory mediators. Western blotting analysis was performed to identify the proinflammatory mediators (TNF α, p38 MAPK), anti-inflammatory cytokine IL 10 and apoptotic marker proteins (Caspase 3, Bcl-2 and Bax) in brain tissue. Prophylactic treatment with trans anethole significantly down regulated the expressions of TNF-α, caspase 3, Bax, dephosphorylated p38 MAPK and upregulated the expressions of Bcl-2, IL 10 in a dose dependent manner in comparison with disease control group. Conclusion: An anti-inflammatory and anti- apoptotic activity of transanethole protected from cerebral ischemia reperfusion injury.

Trans-anethole Ameliorates Intestinal Injury Through Activation of Nrf2 Signaling Pathway in Subclinical Necrotic Enteritis-Induced Broilers.

This study was conducted to investigate the alleviative effects of trans-anethole (TA) on intestinal oxidative stress by enhancing the activities of intestinal antioxidant enzymes and activating the Nrf2 signaling pathway in subclinical necrotic enteritis (NE) infected broilers. A total of 192 1-day-old male Arbor Acres broilers were randomly allocated into three treatment groups: (1) control (CON); (2) subclinical NE challenge (NE); (3) NE challenge + 600 mg/kg TA (NE+TA600). Subclinical NE was induced by oral administration of live coccidiosis vaccine containing 2 × 104 oocysts at 10 days of age and 2 ml of Clostridium perfringens type A solution (3 × 108 CFU/ml) daily from days 14 to 19. The results showed that NE infection led to a severe decline (p < 0.05) in the final body weight (BW) and average daily gain (ADG), but an increase (p < 0.05) in feed/gain (F/G) of broilers at day 10–21 and day 1–21 compared with the control group. TA administration improved (p < 0.05) the growth performance of NE birds. The intestinal villus height (VH) and villus height/crypt depth (VH/CD) were reduced (p < 0.05) by NE challenge as compared with those of the control group, which was elevated by TA administration. Subclinical NE infection decreased (p < 0.05) serum activities of total superoxide dismutase (T-SOD), total antioxidant capacity (T-AOC), and jejunal and ileal glutathione peroxidase (GSH-PX), and T-SOD activity as well as T-AOC in the jejunum, while TA interventions positively elevated that (p < 0.05). Administration of TA protected the intestine against oxidative stress through up-regulation of intestinal nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway as compared with the NE group (p < 0.05). In addition, dietary inclusion of TA elevated (p < 0.05) mRNA abundance of c-mesenchymal-epithelial transition factor (c-Met), jejunal epidermal growth factor receptor (EGFR), and transforming growth factor-beta 1 (TGF-β1) in the jejunum and ileum of birds after subclinical NE challenge. In conclusion, 600 mg/kg of TA may be a promising tool to prevent and control subclinical NE by increasing intestinal antioxidant status in broilers.

Effects of dietary supplementation of trans-anethole on the intestinal antioxidant status, immune function, and liver lipid metabolism in broilers

This study was aimed to investigate the effects of trans-anethole (TA) on intestinal antioxidant status, immune function and liver lipid metabolism in broilers. A total of 256 1-day-old Arbour Acres broilers were allocated to 4 treatment groups with 8 birds per replicate. TA was added to diets at a concentration of 0 (control), 400, 600, and 800 mg/kg for 42 days. Dietary supplementation of TA increased the average daily feed intake (ADFI) of birds throughout the entire period. TA supplemented at 400 mg/kg of diet contributed to the lowest jejunal and ileal malondialdehyde (MDA) concentration on d 42, and the highest ileal total superoxide dismutase (T-SOD) activity on d 21 and d 42. TA supplementation at the concentration of 400 and 600 mg/kg of diet increased mRNA expression of ileal interleukin (IL)-4 on d 21, and intestinal secretory immunoglobulin A (sIgA) concentration on d 42. Supplementation of TA at the concentration up to 800 mg/kg of diet had adverse effects on intestinal antioxidant status, immune function and liver lipid metabolism compared with control group. Differently, TA supplemented at 400 mg/kg led to lower mRNA expression of liver ACC1, FASN, sterol regulatory element-binding protein 1c (SREBP-1), and higher CPT1 on d 42. In conclusion, TA inclusion improved intestinal antioxidant status and immune function, and enhanced liver lipid metabolism of broilers. HIGHLIGHTS Supplementation of 400 mg/kg of TA increased intestinal antioxidant status and immune function, and promoted liver lipid metabolism of broilers compared with non-supplemented group.

Preliminary evaluation of selected inflammatory cytokine gene expression in lymphocytes isolated from whole human blood infected with trans-anethole-treated Staphylococcus aureus Newman strain.

In our previous study based on a whole-blood model of sepsis infected with trans-anethole (TA)-treated Staphylococcus aureus, we have found that innate immune response was more effective in comparison to non-treated cells. Due to the previous observation, in the current preliminary study, a primary adaptive immune response was analysed. This study was conducted to evaluate the expression of selected cytokine (IL1B, IL2, IL6, IL10, TNF, TGFB1, IFNG) and Toll-like receptor (TLR2) genes in lymphocytes isolated from whole human blood infected with S. aureus Newman strain treated with TA. The lymphocytes were isolated by density gradient centrifugation from blood samples infected with S. aureus, as well as from non-infected samples. Gene expression was measured using quantitative real-time PCR. The lymphocytes isolated from the blood infected with TA-treated staphylococcal cells demonstrated significantly greater IL10, IL1B, IL6, TNF and TLR2 expression. Hence, it is possible that the previously observed changes in the surface structure of TA-treated S. aureus Newman strain may significantly increase the relative expression of IL10, IL1B, IL6, TNF and TLR2 genes in lymphocytes; however, further studies are needed.

Trans-Anethole Alleviates Subclinical Necro-Haemorrhagic Enteritis-Induced Intestinal Barrier Dysfunction and Intestinal Inflammation in Broilers.

This study investigated the alleviative potential of trans-anethole (TA) on the impaired intestinal barrier and intestinal inflammation and its regulatory effects on gut microbiota in broilers with subclinical necro-hemorrhagic enteritis (NE) challenge. Subclinical NE challenge led to a severe decline in the 21-day body weight (BW) and average daily gain (ADG), but an increase in feed conversion ratio (FCR) and intestinal lesion score of birds compared with controls (P < 0.05). Compared with the subclinical NE group, the TA administration group exhibited lower (P < 0.05) intestinal lesion score and crypt depth (CD), serum diamine oxidase activity, and D-lactate concentration, but higher (P < 0.05) intestinal tight junction protein expressions, villus height (VH), VH/CD, and numbers of proliferating cell nuclear antigen (PCNA)-positive cells. The administration of TA also inhibited (P < 0.05) the expression of intestinal pro-inflammatory cytokines including interleukin (IL)-1β, IL-8, interferon-gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) but increased (P < 0.05) jejunal IL-10 and secretory immunoglobulin A (sIgA) concentration. TA inclusion also led to a remarkable reduction of intestinal NF-kappa-B inhibitor alpha (IκBα) degradation and nuclear factor kappa beta (NF-κB) translocation. Moreover, TA modulated the cecal microbiota abundance and diversity of NE birds, as confirmed by reducing the phylum Firmicutes and genera Ruminococcaceae_UCG-014, Eubacterium_coprostanoligenes_group, and Ruminococcaceae_NK4A214_group when supplemented at 600 mg/kg and reducing genera Butyricicoccus, Oscillibacter, and Flavonifractor when supplemented at 400 mg/kg (P < 0.05). Supplementation of TA in broiler diets could alleviate subclinical NE infection by restoring intestinal barrier integrity, inhibiting NF-κB signaling pathway, and modulating gut microbiota. A 600-mg/kg dose of TA is the optimum concentration for ameliorating subclinical NE in broilers.

Foeniculum vulgare Miller, a New Chemotype from Montenegro.

Previous studies relating to prolonged and fractionated distillation procedures highlighted essential oils’ (EOs) chemical composition to be significantly dependent on the extraction duration and harvesting time. As a continuation, a hydrodistillation procedure was applied to ripe fruit material of fennel, Foeniculum vulgare Miller (Apiaceae), collected from three localities in Montenegro (Podgorica, Nikšić, and Kotor) to furnish a total of 12 EOs. Liquid and vapor phases of the samples were analyzed by Gas Chromatography/Mass Spectrometry and Headspace-Gas Chromatography/Mass Spectrometry techniques, and 18 compounds have been identified. Although both quantitative and qualitative differences between the samples were notable, the phenylpropanoids anethole (ANE) and estragole and the monoterpenoids α-terpineol (TER) and fenchone (FEN) could be singled out as the most abundant constituents. The EOs from Podgorica belong to the most common ANE-rich chemotype, while the predominance of the monoterpenoid fraction is characteristic of the samples from Nikšić and Kotor. The latter is particularly rich in TER (up to 56.5%), with significant amounts of FEN and ANE. This chemical profile could represent a new chemotype of fennel EO. Vapor phases contained mainly monoterpenoids, with increased amounts of FEN and TER, while the number of phenylpropanoids was significantly decreased.

Influence of Trans-anethole on the nutrient digestibility and intestinal barrier function in broilers

This experiment was undertaken to investigate the effects of dietary trans-anethole (TA) at 5 levels (0, 200, 400, 600, and 800 mg/kg of diet) on the growth performance, apparent nutrient digestibility and intestinal barrier function in broilers. Three hundred twenty 1-day-old Arbor Acres broilers were randomly divided into the 5 dietary treatments with 8 replicates each for 42 d. Dietary TA supplementation increased (P < 0.05) average daily feed intake (ADFI), but had no effects (P > 0.05) on average daily gain (ADG), feed/gain (F/G), and body weight (BW) of broilers throughout the entire experimental period. The apparent metabolizable energy (AME) and nitrogen-corrected apparent metabolizable energy (AMEn), the apparent total tract digestibility of dry matter (DM), crude protein (CP), organic matter (OM), and gross energy (GE) showed a quadratic increase (P < 0.05) with the increasing TA concentration in the diet. The apparent ileal digestibility of Lys, Met, Leu, Thr, Ala, Tyr, and Pro were higher (P < 0.05) in birds fed TA diets compared with control group. Dietary supplementation of 400 mg/kg of TA increased (P < 0.05) mRNA levels of jejunal and ileal Na+/glucose co-transporter (SGLT1) on d 21 and d 42, oligopeptide transporter 1 (PepT1) on d 42, and ileal mRNA expressions of occludin (OCLN), claudin-1 (CLDN-1), and mucin 2 (MUC2), villus height (VH), crypt depth (CD), and VH:CD on d 21, as well as jejunal zonula-occludens-1 (ZO-1) and ileal mucin 2 on d 42. Linear or quadratic responses of the jejunal CD and villus VH:CD ratio occurred (P < 0.01) with increasing dietary TA concentration on d 42. The inclusion of 400 mg/kg TA decreased (P < 0.05) cecal Escherichia coli population on d 21 and d 42, but increased (P < 0.05) Bifidobacterium population on d 21 and ileal Bifidobacterium on d 42. In conclusion, 400 mg/kg of TA is the optimum concentration for increasing nutrient utilization and intestinal barrier function of broilers.

Effects of trans-anethole supplementation on serum lipid metabolism parameters, carcass characteristics, meat quality, fatty acid, and amino acid profiles of breast muscle in broiler chickens

This study investigated the effects of trans-anethole (TA) supplementation on the carcass characteristics, meat quality, fatty acid, and amino acid profiles of breast muscle in broilers. A total of 40 one-day-old male broiler chicks (Arbor Acres) were randomly allocated to 5 treatments, respectively, fed a corn-soybean basal diet supplemented with 0 (control), 200, 400, 600, and 800 mg TA/kg diet for 42 d. 600 mg/kg of TA supplementation decreased (P < 0.05) serum triglycerides (TG) on d 21 and d 42, and high density lipoprotein cholesterol (HDL-C) concentration on d 21, but increased (P < 0.01) serum HDL-C concentration on d 42. Dietary supplementation of TA increased (P < 0.01) the half chamber rate (HCR) and eviscerated rate (ER) of broilers. The drip loss (storing 24 and 48 h) and cooking loss of breast muscle in 600 mg/kg TA groups were lower (P < 0.05) than those in control group. The concentration of palmitoleic acid, daturic acid, oleic acid, linoleic acid, α-Linolenic acid, eicostrienoic acid, and pentosapentanoic acid (EPA), MUFA, and PUFA in the breast muscle were higher (P < 0.05) in the 600 mg/kg of TA group compared with other groups. Dietary inclusion of 600 mg/kg of TA also increased (P < 0.05) the concentration of Met, Thr, Asp, Ser, and Glu in breast muscle, tended to increase (P = 0.069) the Lys concentration. In conclusion, results indicated that TA inclusion improved the lipid metabolism, meat quality, fatty acid composition, and amino acid profile of breast muscle in broilers.

Catalytic Stereoselective Conversion of Biomass-Derived 4′-Methoxypropiophenone to Trans-Anethole with a Bifunctional and Recyclable Hf-Based Polymeric Nanocatalyst

Anethole (AN) is widely used as an odor cleaner in daily necessities, and can also be applied in the fields of food additives, drug synthesis, natural preservatives, and polymeric materials’ preparation. Considering environmental and economic benefits, the use of biomass raw materials with non-precious metal catalysts to prepare high-value fine chemicals is a very promising route. Here, we developed an acid-base bifunctional polymeric material (PhP-Hf (1:1.5)) composed of hafnium and phenylphosphonate in a molar ratio of 1:1.5 for catalytic conversion of biomass-derived 4′-methoxypropiophenone (4-MOPP) to AN via cascade Meerwein–Pondorf–Verley (MPV) reduction and dehydration reactions in a single pot. Compared with the traditional catalytic systems that use high-pressure hydrogen as a hydrogen donor, alcohol can be used as a safer and more convenient hydrogen source and solvent. Among the tested alcohols, 2-pentanol was found to be the best candidate in terms of pronounced selectivity. A high AN yield of 98.1% at 99.8% 4-MOPP conversion (TOF: 8.5 h−1) could be achieved over PhP-Hf (1:1.5) at 220 °C for 2 h. Further exploration of the reaction mechanism revealed that the acid and base sites of PhP-Hf (1:1.5) catalyst synergistically promote the MPV reduction step, while the Brønsted acid species significantly contribute to the subsequent dehydration step. In addition, the PhP-Hf polymeric nanocatalyst can be recycled at least five times, showing great potential in the catalytic conversion of biomass.

Network pharmacology-based evaluation of natural compounds with paclitaxel for the treatment of metastatic breast cancer

Metastatic breast cancer is a prevalent life-threatening disease. Paclitaxel (PTX) is widely used in metastatic breast cancer therapy, but the side effects limit its chemotherapeutic application. Multidrug strategies have recently been used to maximize potency and decrease the toxicity of a particular drug by reducing its dosage. Therefore, we have evaluated the combined anti-cancerous effect of PTX with tested natural compounds (andrographolide (AND), silibinin (SIL), mimosine (MIM) and trans-anethole (TA)) using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, trypan blue dye exclusion assay, proliferating cell nuclear antigen (PCNA) staining, network pharmacology, molecular docking, molecular dynamics (MD) and in vivo chick chorioallantoic membrane (CAM) angiogenesis assay. We observed a reduction in the IC50 value of PTX with tested natural compounds. Further, the network pharmacology-based analysis of compound-disease-target (C-D-T) network showed that PTX, AND, SIL, MIM and TA targeted 55, 61, 56, 31 and 18 proteins of metastatic breast cancer, respectively. Molecular docking results indicated that AND and SIL inhibited the C-D-T network's core target kinase insert domain receptor (KDR) protein more effectively than others. While MD showed that the binding of AND with KDR was stronger and more stable than others. In trypan blue dye exclusion assay and PCNA staining, AND and SIL along with PTX were found to be more effective than PTX alone. CAM assay results suggested that AND, SIL and TA increase the anti-angiogenic potential of PTX. Thus, natural compounds can be used to improve the anti-cancer potential of PTX.

Anethole Attenuates Enterotoxigenic Escherichia coli-Induced Intestinal Barrier Disruption and Intestinal Inflammation via Modification of TLR Signaling and Intestinal Microbiota.

This study aimed to investigate the effects of dietary anethole supplementation on the growth performance, intestinal barrier function, inflammatory response, and intestinal microbiota of piglets challenged with enterotoxigenic Escherichia coli K88. Thirty-six weaned piglets (24 ± 1 days old) were randomly allocated into four treatment groups: (1) sham challenge (CON); (2) Escherichia coli K88 challenge (ETEC); (3) Escherichia coli K88 challenge + antibiotics (ATB); and (4) Escherichia coli K88 challenge + anethole (AN). On day 12, the piglets in the ETEC, ATB, and AN group were challenged with 10 mL E. coli K88 (5 × 109 CFU/mL), whereas the piglets in the CON group were orally injected with 10 mL nutrient broth. On day 19, all the piglets were euthanized for sample collection. The results showed that the feed conversion ratio (FCR) was increased in the Escherichia coli K88-challenged piglets, which was reversed by the administration of antibiotics or anethole (P < 0.05). The duodenum and jejunum of the piglets in ETEC group exhibited greater villous atrophy and intestinal morphology disruption than those of the piglets in CON, ATB, and AN groups (P < 0.05). Administration of anethole protected intestinal barrier function and upregulated mucosal layer (mRNA expression of mucin-1 in the jejunum) and tight junction proteins (protein abundance of ZO-1 and Claudin-1 in the ileum) of the piglets challenged with Escherichia coli K88 (P < 0.05). In addition, administration of antibiotics or anethole numerically reduced the plasma concentrations of IL-1β and TNF-α (P < 0.1) and decreased the mRNA expression of TLR5, TLR9, MyD88, IL-1β, TNF-α, IL-6, and IL-10 in the jejunum of the piglets after challenge with Escherichia coli K88 (P < 0.05). Dietary anethole supplementation enriched the abundance of beneficial flora in the intestines of the piglets. In summary, anethole can improve the growth performance of weaned piglets infected by ETEC through attenuating intestinal barrier disruption and intestinal inflammation.

Trans-Anethole attenuated renal injury and reduced expressions of angiotensin II receptor (AT1R) and TGF-β in streptozotocin-induced diabetic rats

Fibrosis is a pathological process in diabetic nephropathy that causes renal failure and dysfunction. Given the known anti-diabetic effects of trans-Anethole (TA), we aimed to investigate its renoprotective and anti-fibrotic effect alone and in combination with losartan in diabetic nephropathy. Male Wistar rats received a single intraperitoneal injection of 65 mg/kg streptozotocin (STZ) for diabetes induction. Diabetic rats were treated orally with saline, TA (80 mg/kg), losartan (Los; 10 mg/kg), or the combination of TA and losartan (TA-Los) daily for five weeks. Renal function was monitored during the study, and renal fibrosis, oxidative stress markers, apoptotic cells, and the expression and localization of AT1R, TGF-β1, and Col-IV were detected in the kidney. Results showed that TA alone and in combination with losartan was able to decrease blood glucose, urea, and creatinine levels and improve kidney function parameters. TA, Los, and TA-Los significantly reduced tubule vascular degeneration, glomerular and tubulointerstitial sclerosis, oxidative stress, and apoptotic cells. Immunohistochemistry analyses showed that TA, losartan, and TA-losartan combination downregulated the AT1R, Col IV, and TGF-β1 expression and distribution in diabetic rat kidneys. Results suggest that TA is able to suppress diabetic nephropathy in rats effectively, probably by decreasing blood glucose levels and downregulating AT1R and TGF-β1 expression.

The hepatoprotective effects of fennel seeds extract and trans-Anethole in streptozotocin-induced liver injury in rats.

Hypoglycemic, anti‐inflammatory, and antioxidant activities of fennel have been recorded in numerous investigations. The study aimed to evaluate the protective effects of fennel or its active component trans‐Anethole (TA) on streptozotocin‐induced liver injury in rats. Rats were injected with a single dose of STZ (65 mg/kg) and treated with fennel (200 and 400 mg/kg), TA (80 mg/kg), or metformin (300 mg/kg) for 35 days. Serum lipid profile and liver enzyme activity (aminotransferases), oxidative stress markers, and the degree of fibrosis in the liver tissue were assessed. Both fennel and TA decreased blood glucose levels, reduced liver enzyme activity, food, and water intake, and intensity of weight loss, reduced serum triglycerides (TG), total cholesterol (TC), low‐density lipoprotein cholesterol (LDL‐c), and increased high‐density lipoprotein cholesterol (HDL‐c). Additionally, fennel and TA significantly reduced MDA concentration while increased CAT activity and thiol content and reduced the degree of injury and fibrosis in the liver of diabetic rats. Our results suggest that fennel seed extract and its active compound TA are able to protect the liver against diabetes‐induced hepatic injury in rats, probably via hypoglycemic and antioxidant effects.

Modulation of Lipid Metabolism by Trans-Anethole in Hepatocytes.

Non-alcoholic fatty liver disease is caused by excessive lipid accumulation in hepatocytes. Although trans-anethole (TAO) affects hypoglycemia and has anti-immune activity and anti-obesity effects, its role in non-alcoholic fatty liver disease remains unknown. This study aimed to evaluate the effects of TAO on cellular senescence, lipid metabolism, and reinforcement of microenvironments in HepG2 cells. To analyze the lipid metabolic activity of TAO, PCR analysis, flow-cytometry, and Oil Red O staining were performed, and mitochondrial membrane potential (MMP) and cellular senescence kits were used for assessing the suppression of cellular senescence. At 2000 μg/mL TAO, the cellular viability was approximately 99%, and cell senescence decreased dose-dependently. In the results for MMP, activity increased with concentration. The levels of lipolytic genes, CPT2, ACADS, and HSL, strongly increased over 3 days and the levels of lipogenic genes, ACC1 and GPAT, were downregulated on the first day at 1000 μg/mL TAO. Consequently, it was found that TAO affects the suppression of cellular senescence, activation of lipid metabolism, and reinforcement of the microenvironment in HepG2 cells, and can be added as a useful component to functional foods to prevent fatty liver disease and cellular senescence, as well as increase the immunoactivity of the liver.

Determination of Trans-Anethole in Essential Oil, Methanolic Extract and Commercial Formulations of Foeniculum vulgare Mill Using a Green RP-HPTLC-Densitometry Method

Due to the lack of ecofriendly/green reversed-phase high-performance thin-layer chromatography (RP-HPTLC) methods for trans-anethole (TAL) and its simplicity over routine analytical techniques, there was a necessity to establish a suitable HPTLC methodology for the quantitative analysis of TAL. Therefore, the first objective of this research was to develop an accurate, rapid and green RP-HPTLC densitometry methodology for the quantitative analysis of TAL in essential oil, traditional and ultrasound-assisted extracts of Foeniculum vulgare Mill and commercial formulations. The second objective was to compare the traditional method of extraction of TAL with its ultrasound-assisted method of extraction. The chromatogram of TAL from essential oil and traditional and ultrasound-assisted extracts of fennel and commercial formulations was verified by recoding its single spectra at Rf = 0.31 ± 0.01 in comparison to standard TAL. The proposed analytical methodology has been found to be superior in terms of linearity, accuracy and precision compared to most of the reported analytical methods for TAL analysis. The amount of TAL in the essential oil of fennel was recorded as 8.82 mg per g of oil. The content of TAL in traditional extracts of fennel, formulation 1 (dietary supplement 1) and formulation 2 (dietary supplement 2), was recorded as 6.44, 4.88 and 4.48 mg per g, respectively. The amount of TAL in ultrasound-assisted extracts of fennel, formulation 1 and formulation 2, was recorded as 8.34, 6.46 and 5.81 mg per g, respectively. The ultrasound method of extraction of TAL was found to be better than the traditional method of extraction. The results of validation studies and phytochemical analysis showed that the proposed methodology could be efficiently utilized for the quantification of TAL in the wide range of products having TAL as a component.

Anti-inflammatory Effect of Low-Dose Anethole and Ibuprofen Combination Is Accompanied by Partial Prevention of Hepatic Metabolic Changes in Arthritic Rats.

Anethole (AN) is a natural compound that has attracted great scientific interest because of its numerous biological activities, including anti-inflammatory effects. However, these effects were obtained with high doses of AN, which may be one limitation of its therapeutic use. This study evaluated the effects of a low-dose AN and ibuprofen (IB) combination on inflammatory parameters in Freund's complete adjuvant-induced arthritis (AIA) and arthritis-induced hepatic metabolic changes. Holtzman rats were used and divided into groups: normal, AIA (control), arthritics treated with IB, arthritics treated with AN, and arthritics treated with AN + IB. The volume of the paws, the appearance of secondary lesions, and the number of synovial leukocytes were evaluated. Gluconeogenesis and ureagenesis from alanine were determined in the rat liver in isolated perfusion. The AN + IB (62.5 + 8.75mg/kg) treatment exerted an inhibitory effect on inflammatory parameters and partially prevented hepatic metabolic changes that was similar to the effect of high-dose IB (35mg/kg) and AN (250mg/kg) treatment. This effect of the treatments on hepatic metabolism can be, partly at least, explained by the preservation of both the alanine aminotransferase (ALT) activity and the cytosolic NADH/NAD+ redox potential in the liver. Taken together, the data obtained provided evidence that the AN + IB combination at lower doses than AN and IB treatment alone had beneficial inhibitory potential for the treatment of AIA and attenuated metabolic changes in the liver. Graphical Abstract.

Insecticidal Efficacy and Repellency of Trans-Anethole Against Four Stored-Product Insect Pests

In this study, it was investigated that repellency and insecticidal efficacy of trans-anethole of botanical origin on major stored product on pests species, namely Sitophilus granarius (L.) (Coleoptera: Curculionidae), Sitophilus oryzae (L.) (Coleoptera: Curculionidae), Tribolium confusum Jacquelin du Val (Coleoptera, Tenebrionidae) and Tribolium castaneum (Herbst) (Coleoptera, Tenebrionidae). Repellency effect was evaluated by choice test in petri dishes, while the mortality effect was examined by exposure to wheat treated at different concentrations, such as 1, 2, 3, 4, and 8 μL trans anethole. In efficacy tests, after 72 hours exposure, the highest adult mortality was found on T. castaneum with a 60% mortality, while other test species showed no significant mortality. On the other hand, repellency tests revealed varying degree of repellency depending on the application dose of trans-anethole. It was determined that S. granarius belongs to repellent class III, while all of the other species fall under repellent class IV. F1 progeny decreased as trans-anethole concentration increased, and in this context 8 μL of trans-anethole is proved to be the optimal concentration causing maximum decrease in progeny production. Among the insect species tested, T. confusum was found to be the most sensitive to trans-anethole with 100% decrease in F1 progeny production. Our results indicate that trans-anethole can be used as a potential repellent for the control of major stored grain pests. Additionally, trans-anethole, by its contact efficacy, might be considered as a grain protectant against S. granarius, S. oryzae, T. confusum and T. castaneum.