Anemonin

Stability of protoanemonin in plant extracts from Helleborus niger L. and Pulsatilla vulgaris Mill.[Pubmed:32497949]

J Pharm Biomed Anal. 2020 Sep 5;188:113370.

The concentration levels and stability of protoAnemonin, a characteristic constituent of Ranunculaceae species with antimicrobial and fungicidal properties, were studied for the first time in plant extracts prepared from Helleborus niger L. and Pulsatilla vulgaris Mill. using fermentative production processes. ProtoAnemonin levels quantified by HPLC-DAD analysis were 0.0345 and 0.0662 mg/g in two freshly prepared Helleborus (whole, flowering plant) extracts and 0.3875 mg/g and 0.4193 mg/g in Pulsatilla (flowers) extracts. ProtoAnemonin proved to be rather instable in aqueous-fermented extracts stored at 15 degrees C in the dark, and its concentration decreased rapidly over 12 months of storage independently of the plant species. The decrease was most pronounced when initial concentrations were high (decrease by about 70%). In contrast, low protoAnemonin levels remained stable in solution for more than 12 months. Anemonin, the dimer of protoAnemonin, was detected in increasing concentrations only in Pulsatilla samples, but its concentration only accounted for less than 50% of the theoretically expected amount. With respect to fermented extracts, both physical processes such as self-polymerization and adsorption/binding to other extract constituents as well as biodegradation were concluded to be responsible for protoAnemonin decline. As opposed to plant extracts, both protoAnemonin and Anemonin levels decreased in 0.22 mum-filtered samples stored in vials. This may be explained by a reduced release from plant material in combination with physicochemically induced degradation. Reduction was most pronounced upon light exposure and elevated temperatures, clearly indicating that photochemical degradation is involved. Contents of protoAnemonin in a set of extract batches were 0.0896 +/- 0.0125 mg/g and 0.0618 +/- 0.0180 mg/g in Helleborus and Pulsatilla extracts, and Anemonin levels were 0.0230 +/- 0.0076 mg/g and 0.0482 +/- 0.0282 mg/g, respectively. Due to its antibiotic effects, but also its reactivity, protoAnemonin is a therapeutically and toxicologically relevant constituent, and its concentration should therefore be carefully monitored.

Anemonin Attenuates RANKL-Induced Osteoclastogenesis and Ameliorates LPS-Induced Inflammatory Bone Loss in Mice via Modulation of NFATc1.[Pubmed:32116686]

Front Pharmacol. 2020 Feb 10;10:1696.

Osteoporosis is a metabolic bone disease characterized by insufficient osteoblastic function and/or excessive osteoclastic activity. One promising strategy for treating osteoporosis is inhibiting excessive osteoclast resorbing activity. Previous studies have revealed that Anemonin (ANE), isolated from various types of Chinese natural herbs, has anti-inflammatory and anti-oxidative properties. However, whether ANE regulates osteoclastogenesis is unknown. This study aimed to investigate the potential effect of ANE on osteoclastogenesis and inflammatory bone loss in mice. In in vitro studies, ANE suppressed RANKL-stimulated osteoclast differentiation and function by downregulating the expression of osteoclast master transcriptor NFATc1, as well as its upstream transcriptor c-Fos, by decreasing NF-kappaB and ERK1/2 signaling. Interestingly, ANE did not change the phosphorylation and degradation of IkappaB-alpha and activation of JNK and p38 MAPKs. However, ANE repressed the phosphorylation of MSK-1 which is the downstream target of ERK1/2 and p38 MAPK and can phosphorylate NF-kappaB p65 subunit. These results implicated that ANE might suppress NF-kappaB activity via modulation of ERK1/2 mediated NF-kappaB phosphorylation. In addition, ANE directly suppressed NFATc1 transcription by inhibiting Blimp-1 expression, and the subsequent enhancement of the expression of NFATc1 negative regulators, Bcl-6 and IRF-8. Moreover, in vivo studies were conducted using an LPS-induced inflammatory bone loss mice model. Micro-CT and histology analysis showed that ANE treatment significantly improved trabecular bone parameters and bone destruction. These data indicate that ANE can attenuate RANKL-induced osteoclastogenesis and ameliorate LPS-induced inflammatory bone loss in mice through modulation of NFATc1 via ERK1/2-mediated NF-kappaB phosphorylation and Blimp1 signal pathways. ANE may provide new treatment options for osteoclast-related diseases.

Regulation of iRhom-2/Tumor Necrosis Factor-alpha Converting Enzyme Pathway and Oxidative Stress Protects the Renal Injury with Anemonin in Streptozotocin-Induced Diabetic Nephropathy Neonatal Rat Model.[Pubmed:31344701]

Pharmacology. 2019;104(5-6):258-266.

Diabetic nephropathy (DN) is a chronic complication of diabetes, and thus the present investigation evaluates the nephroprotective effect of Anemonin against streptozotocin (STZ)-induced DN rats. Diabetes was induced by intraperitoneal administration of STZ (50 mg/kg) on day 2 and 3 postnatal, and rats were kept as such for the duration of 12 weeks. Thereafter, rats were treated with Anemonin 75 and 150 mg/kg per oral for the period 4 week which means between the period of 12-16 weeks. Effect of Anemonin was estimated by determining the blood glucose, markers of nephropathy, and mediators of inflammation in the serum and activity of tumor necrosis factor-alpha (TNF-alpha)converting enzyme (TACE) in the kidney tissue of DN rats. Moreover, reverse transcriptase polymerase chain reaction and western blot assay were determined in the kidney tissue homogenate of DN rats. Histopathology study was done by Periodic acid-Schiff's and masson staining for the pathological changes and apoptosis of podocytes in the kidney tissue of DN rats. Moreover, production of reactive oxygen species (ROS) was estimated in the kidney tissue by 2',7'-dichlorofluorescein staining. Data of study reveal that Anemonin significantly reduces (p < 0.01) the blood glucose and markers of renal injury in the serum and urine of DN rats. There was a reduction in the level of cytokines in the serum, and production of ROS and activity of TACE were reduced in the kidney tissue of the Anemonin-treated group than in the DN group. Expression of iRhom-2, TACE, TNF-alpha, and inducible nitric oxide synthase protein and histopathology of kidney tissue were attenuated in the Anemonin-treated group in DN rats. In conclusion, data of study reveal that treatment with Anemonin ameliorates progression of renal injury by regulating TACE/iRhom-2 signaling pathway.

Tunisian Clematis flammula Essential Oil Enhances Wound Healing: GC-MS Analysis, Biochemical and Histological Assessment.[Pubmed:30404969]

J Oleo Sci. 2018;67(11):1483-1499.

The aerial part of Clematis flammula (Ranunculaceae) has been traditionally used in the treatment of skin diseases including mycotic infection in the Tunisian traditional medicine. The study was undertaken to extract and determine the essential oil chemical composition of Clematis flammula aerial parts and to assess the potential of Anemonin in wound healing on mechanically wounded wistar rats. The essential oil was obtained by hydrodistillation and analyzed by GC-MS. Anemonin was isolated and then incorporated as active in a cream for which the cytotoxicity was evaluated by methyl thiazolyl tetrazolium (MTT)-based colorimetric assay. Then, its potential in wound healing on mechanically wounded wistar rats was assessed. The GC-MS analysis showed that the major compound was protoAnemonin (86.74%) which spontaneously dimerised in part to form the Anemonin. The wound healing activity of Anemonin cream exhibited a non toxic potential of Anemonin at a concentration of 25 microg/mL with a cell migration efficiency that reaches more than 80% after 48 hours of treatment. Wound healing efficiency was evaluated by monitoring morphological and skin histological analyses. Comparable wound surface reduction of the group treated by Anemonin cream (p >/= 0.05) when compared to the reference treated group. The skin histological analysis showed the completely wound closure. Antioxidant activity was assessed by the malondialdehyde (MDA) rates and antioxidant enzymes (glutathione peroxidase (GPx) and catalase) determination. The results provided strong support for the effective wound healing activity of Anemonin cream, making it a promising candidate as a therapeutic agent in tissue repairing processes.

The anemonin content of four different Ranunculus species.[Pubmed:30393208]

Pak J Pharm Sci. 2018 Sep;31(5(Supplementary)):2027-2032.

The Ranunculus species are poorly known as medicinal plants. They have potential toxicity given by the ranunculin and its enzymatic degradation compounds: protoAnemonin and Anemonin. This paper aims to evaluate the Anemonin content of four species: R. bulbosus, R. ficaria, R. sardous and R. sceleratus. The evaluation was performed by TLC and HPLC. There were evaluated two types of extracts hydroalcoholic (HA) and glycerol-ethanol (GE). The most concentrated extract in Anemonin was found to be the R. sardous aerial part HA extract: 2.66 mg/ml. The lowest Anemonin content is in R. sceleratus: 0.13-0.19 mg/ml. In R. bulbosus aerial part the Anemonin content is less than the used HPLC method detection limits (7.68 mg/ml). In all cases the GE extracts are less concentrated in Anemonin, being more safely for human administration.

Antifungal, molluscicidal and larvicidal assessment of anemonin and Clematis flammula L. extracts against mollusc Galba truncatula, intermediate host of Fasciola hepatica in Tunisia.[Pubmed:29111192]

Asian Pac J Trop Med. 2017 Oct;10(10):967-973.

OBJECTIVE: To investigate the potential of Anemonin and Clematis flammula (C. flammula) extracts against infective organisms. METHODS: The molluscicidal activities of Anemonin and C. flammula extracts against Galba truncatula Mull. (Lymnaeidae) and Fasciola hepatica larval stages contaminating this snail in Tunisia were assessed by testing six groups of snails in 250 mL of extracts and aqueous dechlorinated solutions with different concentrations (ranging from 2.5 to 20.0 mg/L) for 48 h. Besides, the antifungal potential of C. flammula leaves and flowers was evaluated by using the diffusion agar and broth dilution methods against four fungal strains: Aspergillus niger, Pythium catenulatum, Rhizoctonia solani and Fusarium phyllophilum. RESULTS: As a result, hexane and ethyl acetate flower extracts exhibited significant molluscicidal activities with LC50 median lethal concentrations values of 11.87 and 11.65 mg/L, respectively while LC50 value of Anemonin was 9.64 mg/L after 48 h exposure. The flower extracts showed a larvicidal effect with a deterioration rate exceeding 35.39% where flower ethyl acetate residue gave a deterioration rate of cercariae close to 97%. Moreover, C. flammula extracts were not noxious to the associated fauna survival. All extracts inhibited the growth of P. catenulatum, the leaves and flowers methanolic extracts had the more important fungicide action with minimum inhibitory concentrations of 1.56 and 3.12 mg/mL together with minimum fungistatic concentrations of 3.12 and 6.25 mg/mL respectively. Only flower extracts were active against Rhizoctonia solani with minimum inhibitory concentrations varying between 0.70 and 1.56 mg/mL and 6.25 mg/mL of minimum fungistatic concentration. Phytochemical tests showed that the antifungal activity may be attributed to the presence of the flavonoids/saponins in the methanolic extracts and the molluscicide effects could be due to the richness of hexane and ethyl acetate extracts on sterols and triterpenoids. CONCLUSIONS: This study emphasizes the important molluscicidal and antiparasitic effects of flower ethyl acetate extracts and Anemonin compound as well as the considerable antifungal activities of methanolic extracts. These results improve the therapeutic virtues of C. flammula aerial part extracts.

Anemonin attenuates osteoarthritis progression through inhibiting the activation of IL-1beta/NF-kappaB pathway.[Pubmed:28643466]

J Cell Mol Med. 2017 Dec;21(12):3231-3243.

The osteoarthritis (OA) progression is now considered to be related to inflammation. Anemonin (ANE) is a small natural molecule extracted from various kinds of Chinese traditional herbs and has been shown to inhibiting inflammation response. In this study, we examined whether ANE could attenuate the progression of OA via suppression of IL-1beta/NF-kappaB pathway activation. Destabilization of the medial meniscus (DMM) was performed in 10-week-old male C57BL/6J mice. ANE was then intra-articularly injected into joint capsule for 8 and 12 weeks. Human articular chondrocytes and cartilage explants challenged with interleukin-1beta (IL-1beta) were treated with ANE. We found that ANE delayed articular cartilage degeneration in vitro and in vivo. In particular, proteoglycan loss and chondrocyte hypertrophy were significantly decreased in ANE -treated mice compared with vehicle-treated mice. ANE decreased the expressions of matrix metalloproteinase-13 (MMP13), A disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5), collagen X (Col X) while increasing Aggrecan level in murine with DMM surgery. ANE treatment also attenuated proteoglycan loss in human cartilage explants treated with IL-1beta ex vivo. ANE is a potent protective molecule for OA; it delays OA progression by suppressing ECM loss and chondrocyte hypertrophy partially by suppressing IL-1beta/NF-kappaB pathway activation.

Anemonin improves intestinal barrier restoration and influences TGF-beta1 and EGFR signaling pathways in LPS-challenged piglets.[Pubmed:27189428]

Innate Immun. 2016 Jul;22(5):344-52.

The present study was aimed at investigating whether dietary Anemonin could alleviate LPS-induced intestinal injury and improve intestinal barrier restoration in a piglet model. Eighteen 35-d-old pigs were randomly assigned to three treatment groups (control, LPS and LPS+Anemonin). The control and LPS groups were fed a basal diet, and the LPS + Anemonin group received the basal diet + 100 mg Anemonin/kg diet. After 21 d of feeding, the LPS- and Anemonin-treated piglets received i.p. administration of LPS; the control group received saline. At 4 h post-injection, jejunum samples were collected. The results showed that supplemental Anemonin increased villus height and transepithelial electrical resistance, and decreased crypt depth and paracellular flux of dextran (4 kDa) compared with the LPS group. Moreover, Anemonin increased tight junction claudin-1, occludin and ZO-1 expression in the jejunal mucosa, compared with LPS group. Anemonin also decreased TNF-alpha, IL-6, IL-8 and IL-1beta mRNA expression. Supplementation with Anemonin also increased TGF-beta1 mRNA and protein expression, Smad4 and Smad7 mRNA expressions, and epidermal growth factor and epidermal growth factor receptor (EGFR) mRNA expression in the jejunal mucosa. These findings suggest that dietary Anemonin attenuates LPS-induced intestinal injury by improving mucosa restoration, alleviating intestinal inflammation and influencing TGF-beta1 canonical Smads and EGFR signaling pathways.

Skin permeation profile and anti-inflammatory effect of anemonin extracted from weilingxian.[Pubmed:27183707]

Pharmazie. 2016 Mar;71(3):134-8.

The aim of this study was to evaluate the skin permeability of Anemonin, which was extracted from the Chinese herb weilingxian, and its potency of relieving the inflammation caused by rheumatoid arthritis (RA). To optimize the formulation, the solubility of Anemonin in water and selected concentration of ethanol-water vehicles was determined. The effect of ethanol on the permeation of Anemonin through human skin was then studied. Additionally, the influences of hydroxypropyl methylcellulose E50 (HPMC) and Carbomer 934 in different concentrations on the permeation of drug were investigated. Finally, the anti-inflammatory effect of the optimized formulation was assessed by murine model of xylene-induced ear edema. The results showed that the solubility and transdermal permeation of Anemonin in ethanol-water vehicles linearly depended on the ethanol concentration. The combination of 30% ethanol and 3% Azone had a synergistic enhancement effect and was therefore selected for gel preparation. The 0.14% Anemonin gel prepared with 1% HPMC exhibited the highest transdermal flux. The xylene-induced ear edema inhibitory rate of the optimized formulation was 48.85%. The results indicated that transdermal administration of Anemonin is a potential modality for combating inflammation caused by RA.

Antioxidant C-glycosylflavones of Drymaria cordata (Linn.) Willd.[Pubmed:26642770]

Arch Pharm Res. 2016 Jan;39(1):43-50.

A new C-glycosylflavone, drymaritin E (6-C-(3-keto-beta-digitoxopyranosyl)-4'-O-(beta-D-glucopyranosyl)-7-methoxyl-5,4' -dihydroxylflavone) 1 was isolated from the oily upper phase (SU) of the MeOH extract from aerial parts of Drymaria cordata together with two known compounds (cassiaoccidentalin A 2 and Anemonin 3) and an inseparable mixture of two known C-glycosylflavones 5,4'-dihydroxy-7-methoxyflavone-6-C-(2''-O-alpha-L-rhamnopyranosyl)-beta-D-glucop yranoside 4a and 5,7,3',4'-tetrahydroxyflavone-6-C-(2''-O-alpha-L-rhamnopyranosyl)-beta-D-glucopyr anoside 4b. The alkaline hydrolysis of 3 led to a new hemisynthetic derivative, sodium anemonate (sodium 2-((1'E) 2'-sodium-carboxylate-vinyl)-5-oxo-cyclohex-1-ene carboxylate) 3a. The chemical structures were determined by spectroscopic methods ((1)H NMR, (13)C NMR, (1)H-(1)H COSY, HMBC, HSQC, and NOESY) and mass spectrometry (ESI-MS). C-glycosylflavones had significant free radical-scavenging activities on the radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). However, SU and compounds 3 and 3a exhibited no activity. In particular, compound 1 exhibited a concentration-dependent radical scavenging activity on DPPH with EC50 of 31.43 microg/mL.

Anemonin alleviates nerve injury after cerebral ischemia and reperfusion (i/r) in rats by improving antioxidant activities and inhibiting apoptosis pathway.[Pubmed:24443273]

J Mol Neurosci. 2014 Jun;53(2):271-9.

In the present study, we aimed at evaluating the potential neuroprotective effect and the underlying mechanism of Anemonin against cerebral ischemia and reperfusion (I/R) injury. Anemonin was administered to rats by the intraperitoneally (i.p.) route once daily for 7 days before middle cerebral artery occlusion (MCAO). Focal cerebral ischemia was induced by 90 min of MCAO followed by 24 h of reperfusion. After that, animals were sacrificed by decapitation, brain was removed, and various biochemical estimations, neurological status, and assessment of cerebral infarct size were carried out. MCAO followed by 24 h of reperfusion caused a significant increase in infarct size, neurological deficit score, malondialdehyde (MDA) content, reactive oxygen species (ROS) level, and DNA fragmentation, as well as a decrease in the activities of superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), glutathione peroxidase (GPx), and Na(+), K(+)-ATPase in the brain. Furthermore, elevated Bax expression, increased caspase-3 cleavage, and decreased Bcl-2 expression were observed in nontreated rats in response to focal cerebral I/R injury. However, pretreatment with Anemonin significantly reversed these levels of biochemical parameters, reduced cerebral infarct size, and improved the neurologic score in cerebral ischemic animals. Additionally, a wide distribution of Anemonin in plasma and brain tissues and the brain-to-plasma partition coefficient (Ri) ratio of 0.7 at 90 min indicated that this compound could penetrate the blood-brain barrier (BBB). These results showed that pretreatment with Anemonin provided a significant protection against cerebral I/R injury in rats by, at least in part, its antioxidant action and consequent inhibition of apoptosis.

The genus Clematis (Ranunculaceae): chemical and pharmacological perspectives.[Pubmed:22728167]

J Ethnopharmacol. 2012 Aug 30;143(1):116-50.

ETHNOPHARMACOLOGICAL RELEVANCE: Twenty six species of the genus Clematis (Ranunculaceae) have been traditionally used in various systems of medicine for the treatment of ailments such as nervous disorders, syphilis, gout, malaria, dysentry, rheumatism, asthma, and as analgesic, anti-inflammatory, diuretic, antitumour, antibacterial and anticancer. AIM OF THE REVIEW: To emphasize on ethnopharmacology, chemical constituents, pharmacology, toxicology and clinical studies of various species of the genus Clematis. MATERIALS AND METHODS: The available information on Clematis species was collected through electronic search of major scientific databases. RESULTS: A survey of literature revealed that triterpene saponins, alkaloids, flavonoids, lignans, steroids, coumarins, macrocyclic compounds, phenolic glycosides, Anemonin and volatile oils constitute major classes of chemical constituents in the genus Clematis. Preliminary analgesic, anticancer, anti-inflammatory, diuretic, antiarthritis, hepatoprotective, hypotensive and HIV-1 protease inhibitor activity studies have been carried out on crude extracts of 26 traditionally used and medicinally promising species of Clematis genus. CONCLUSIONS: The species of the genus Clematis emerged as good source of traditional medicine for the treatment of various ailments. Although few experimental studies validated their traditional claims, but employed uncharacterized crude extracts. Such Clematis species need to be explored properly following bioactivity-directed fractionation with a view to isolate bioactive constituents, and to evaluate their possible mode of actions. These species hold great potential for detailed clinical studies so that these could be exploited as potential drugs. The review will help researchers to select medicinally potential species of Clematis genus for future research.

Interleukin-6 and Cyclooxygenase-2 downregulation by fatty-acid fractions of Ranunculus constantinopolitanus.[Pubmed:19917107]

BMC Complement Altern Med. 2009 Nov 16;9:44.

BACKGROUND: Medicinal plants represent alternative means for the treatment of several chronic diseases, including inflammation. The genus Ranunculus, a representative of the Ranunculaceae family, has been reported to possess anti-inflammatory, analgesic, antiviral, antibacterial, antiparasitic and antifungal activities, possibly due to the presence of Anemonin and other. Different studies have shown the occurrence of unusual fatty acids (FAs) in Ranunculaceae; however, their therapeutic role has not been investigated. The purpose of this study is to characterize potential anti-inflammatory bioactivities in Ranunculus constantinopolitanus D'Urv., traditionally used in Eastern Mediterranean folk medicine. METHODS: The aerial part of R. constantinopolitanus was subjected to methanol (MeOH) extraction and solvent fractionation. The bioactive fraction (I.2) was further fractionated using column chromatography, and the biologically active subfraction (Y2+3) was identified using infrared (IR) spectroscopy, nuclear magnetic resonance (NMR) and gas chromatography-mass spectrometry (GC-MS). The effects of I.2 and Y2+3 on cell viability were studied in mouse mammary epithelial SCp2 cells using trypan blue exclusion method. To study the anti-inflammatory activities of I.2 and Y2+3, their ability to reduce interleukin (IL)-6 levels was assessed in endotoxin (ET)-stimulated SCp2 cells using enzyme-linked immunosorbent assay (ELISA). In addition, the ability of Y2+3 to reduce cyclooxygenase (COX)-2 expression was studied in IL-1-treated mouse intestinal epithelial Mode-K cells via western blotting. Data were analyzed by one-way analysis of variance (ANOVA), Student-Newman-Keuls (SNK), Tukey HSD, two-sample t-test and Dunnett t-tests for multiple comparisons. RESULTS: The chloroform fraction (I.2) derived from crude MeOH extract of the plant, in addition to Y2+3, a FA mix isolated from this fraction and containing palmitic acid, C18:2 and C18:1 isomers and stearic acid (1:5:8:1 ratio), reduced ET-induced IL-6 levels in SCp2 cells without affecting cell viability or morphology. When compared to fish oil, conjugated linoleic acid (CLA) and to individual FAs as palmitic, linoleic, oleic and stearic acid or to a mix of these FAs (1:5:8:1 ratio), Y2+3 exhibited higher potency in reducing ET-induced IL-6 levels within a shorter period of time. Y2+3 also reduced COX-2 expression in IL-1-treated Mode-K cells. CONCLUSION: Our studies demonstrate the existence of potential anti-inflammatory bioactivities in R. constantinopolitanus and attribute them to a FA mix in this plant.

Chinese herbal medicinal ingredients inhibit secretion of IL-6, IL-8, E-selectin and TXB2 in LPS-induced rat intestinal microvascular endothelial cells.[Pubmed:19874221]

Immunopharmacol Immunotoxicol. 2009;31(4):550-5.

The aim of the research was to investigate the anti-inflammatory mechanism of Pulsatillae Decoction (PD), the levels of interleukin (IL)-6, IL-8, E-selectin, and thromboxane B(2) (TXB(2)) secreted by cultured rat intestinal microvascular endothelial cells (RIMECs) were determined after treatment with its active ingredients, namely anemoside B4, Anemonin, berberine, jatrorrhizine, palmatine, aesculin, and esculetin. RIMECs were challenged with 1 microg/mL lipopolysaccharide (LPS) for 3 h, and then treated with each of the seven ingredients at three concentrations (1, 5 and 10 microg/mL) for 24 h. The results revealed that Anemonin, aesculin and esculetin inhibited the production of IL-6, aesculin and esculetin inhibited the secretion of IL-8, anemoside B4, berberine and jatrorrhizine downregulated E-selectin expression, Anemonin, berberine, jatrorrhizine and palmatine decreased the content of TXB(2). All these changes were significant. Taken together, the data suggest that all seven active ingredients of PD can effectively reduce inflammatory response, thus relieving intestinal dysfunction via multiple pathways.