Your browser doesn't support javascript.
loading
Montrer: 20 | 50 | 100
Résultats 1 - 20 de 190
Filtre
1.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 47-55, 2023.
Article Dans Chinois | WPRIM | ID: wpr-996809

Résumé

ObjectiveTo investigate the mechanism of icariin in ameliorating efferocytosis dysfunction and inflammatory response of alveolar macrophages induced by cigarette smoke extract via the peroxisome proliferator-activated receptor gamma (PPARγ) signaling pathway. MethodThe untreated rat alveolar macrophages (NR8383) were taken as the blank group. The NR8383 cells treated with 10% cigarette smoke extract were divided into model, low-, medium-, and high-dose (10, 20, 40 μmol·L-1) icariin, PPARγ inhibitor, and PPARγ inhibitor + low-, medium-, and high-dose icariin groups. Alamar blue colorimetry was employed to examine the proliferation and toxicity of icariin on NR8383 cells. The efferocytosis rate of NR8383 cells was detected by flow cytometry. Enzyme-linked immunosorbent assay was employed to measure the levels of tumor necrosis factor-alpha (TNF-α), transforming growth factor-β1 (TGF-β1), and milk fat globule-epidermal growth factor 8 (MFG-E8). Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were employed to determine the protein and mRNA levels, respectively, of PPARγ, CD36, and RAS-related C3 botulinum toxin substrate 1 (Rac1). ResultThe efferocytosis dysfunction model of NR8383 was established with the cigarette smoke extract. Compared with the blank control group, the model group showed decreased efferocytosis rate (P<0.05), elevated TNF-α level (P<0.05), lowered TGF-β1 and MFG-E8 levels (P<0.01), and down-regulated mRNA and protein levels of PPARγ, CD36, and Rac1 (P<0.05, P<0.01). Compared with the model group, the treatment with icariin increased the efferocytosis rate (P<0.05, P<0.01), lowered the TNF-α level (P<0.01), elevated TGF-β1 and MFG-E8 levels (P<0.05), and up-regulated the protein and mRNA levels of PPARγ, CD36, and Rac1 (P<0.05, P<0.01). Compared with icariin alone, PPARγ inhibitor + icariin decreased the efferocytosis rate (P<0.05) and down-regulated the protein and mRNA levels of PPARγ (P<0.05, P<0.01). In addition, PPARγ inhibitor + low-dose icariin down-regulated the protein level of CD36 (P<0.01) and PPARγ inhibitor + low-/medium-dose icariin up-regulated the protein level of Rac1 (P<0.05). ConclusionIcariin ameliorates the cigarette smoke extract-induced efferocytosis dysfunction of alveolar macrophage by regulating the PPARγ signaling pathway and cytoskeletal structure rearrangement.

2.
Chinese Journal of Pharmacology and Toxicology ; (6): 519-520, 2023.
Article Dans Chinois | WPRIM | ID: wpr-992199

Résumé

OBJECTIVE To investigate the effect of icariin(ICA)on the ubiquitination modification of β-amy-loid precursor protein(APP)in Alzheimer's disease mice.METHODS In vitro,① HEK 293 cells stably overex-pressing human APP695(OE-hAPP)were treated with different concentrations of ICA(10-100 μmol·L-1)for 24 h and the cell viability was detected by MTT assay.②CHX(50 mg·L-1)was used to block protein synthesis and MG132(20 μmol·L-1)inhibits proteasome activity,then the level of APP in different time(0,0.5,1,2,3 and 4 h)and the ubiquitination were tested by Western blotting.③ E3 ubiquitin ligases HMG-CoA reductase degradation pro-tein 1(HRD1)protein expression in OE-hAPP was tested by Western blotting,as well as the level and ubiquitination of APP were tested under HRD1 silent condition by Co-IP and Western blotting.In vivo,① male APP/PS1 mice and wild type(WT)mice were randomly divided into 5 groups:WT,WT+ICA,APP/PS1,APP/PS1+ICA,and APP/PS1+donepezil(DPZ)groups.ICA(60 mg·kg-1·d-1)and DPZ(1 mg·kg-1·d-1)were treated for 3 months by gavage from 6 months of age,and WT mice were given equal volume of distilled water.②Morris water maze and Y-maze experiments were used to detect the alteration of spatial learning memory function.③ After then,the brain tissues were collected,total proteins were extracted,APP antibodies were subjected to Co-IP,and total ubiqui-tination(Ub),K48-linked polyubiquitination(UbK48)and K63-linked polyubiquitination of APP level,APP and HRD1 proteins were detected by Western blotting.RESULTS In vitro results showed that ICA significantly enhanced APP degradation(vs control,P<0.01),up-reg-ulated HRD1 expression(vs control,P<0.05;vs OE-hAPP,P<0.05),elevated the level Ub and UbK48 of APP,as well as increased APP degradation.Moreover,silenced HRD1 gene abolished abovementioned effects of ICA(vs control-siRNA,P<0.05;vs HRD1-siRNA,P<0.05).In vivo results showed that ICA improved the spa-tial learning and memory function APP/PS1 mice by Mor-ris water maze and Y-maze tests,increased HRD1 expres-sion(vs APP/PS1 + vehicle,P<0.05),enhanced APP ubiquitination and reduced APP protein level(vs APP/PS1 + vehicle,P<0.01).CONCLUSION ICA promotes the ubiquitination and proteasome-dependent degrada-tion of APP by up-regulating HRD1,thereby improving the spatial learning and memory function of Alzheimer disease mice.

3.
Chinese Journal of Pharmacology and Toxicology ; (6): 515-516, 2023.
Article Dans Chinois | WPRIM | ID: wpr-992196

Résumé

Multiple sclerosis(MS)is a systemic inflammatory illness of the central nervous system that involves demyelinating lesions in the myelin-rich white matter and pathology in the grey matter.Despite signifi-cant advancements in drug research for MS,the dis-ease's complex pathophysiology makes it difficult to treat the progressive forms of the disease.In this study,we identified a natural flavonoid compound icariin(ICA)as a potent effective agent for MS in ameliorating the deterioration of symptoms including the neurological defi-cit score and the body weight in a murine experimental autoimmune encephalomyelitis(EAE)model.These improvements were associated with decreased demyelin-ation in the corpus callosum and neuron loss in the hippo-campus and cortex confirmed by immunohistochemistry analysis.Meanwhile,it was observed that the activation of microglia in cerebral cortex and hippocampus were inhibited followed by the neuroinflammatory cytokines downregulation such as IL-1β,IL-6 and TNF-α after ICA treatment,which was probably attributable to the sup-pression of microglial NLRP3 inflammasome activation.Additionally,molecular docking also revealed the binding force of ICA to NLRP3 inflammasome protein complexes in vitro.Taken together,our findings have demonstrated that ICA,as pleiotropic agent,prevents EAE-induced MS by improving demyelination and neuron loss,which inter-feres with the neuroinflammation via microglial NLRP3 inflammasome activation.

4.
Chinese Journal of Pharmacology and Toxicology ; (6): 513-514, 2023.
Article Dans Chinois | WPRIM | ID: wpr-992194

Résumé

OBJECTIVE To investigate whether icari-in(ICA)plays a neuroprotective role by improving glyco-lytic function through activating Wnt/β-catenin signaling pathway.METHODS HT22 cells were treated with Aβ25-35 for 24 h to establish AD cell model,ICA was added in 2 h before Aβ25-35 and the DKK1(a specific inhibitor of the Wnt signaling pathway)was added in 0.5 h before ICA.Pharmacodynamic study:HT22 cells were divided into control group,ICA group(ICA 10 μmol·L-1),model group(Aβ25-3520 μmol·L-1),model + ICA group(Aβ25-3520 μmol·L-1 +ICA 2.5,5,10 μmol·L-1);Mechanism study:HT22 cells were divided into control group,model group,Aβ25-35+ICA 10 μmol·L-1 group,Aβ25-35+DKK1 group,Aβ25-35+DKK1+ ICA group.The cell viability was detected by MTT assay and the cell morphology was obtained by microscope,the lactate content was detected by lactate assay,the ATP content was measured with the chemiluminescence method,the expression levels of HK1,PKM1 and the pro-tein expression of molecules related to the Wnt/β-catenin signaling pathway(Wnt3a,GSK3β,pGSK3β Try216,pGSK3β Ser9,β-catenin,pβ-catenin Ser33/37 Thr41,Active β-catenin and nuclear β-catenin)was assayed by Western blotting.The nuclear translocation of β-catenin was observed by immunofluorescent staining.RESULTS Compared with the control group,the viability of cells in the model group was reduced,the morphology of cells was significantly damaged,the ATP content and lactate content were significantly decreased,and the glycolytic key enzymes:the protein levels of HK1,PKM1 and the protein levels of Wnt3a,pGSK3β Ser9,active β-catenin and nuclear β-catenin were significantly reduced,and the phosphorylation levels of β-catenin Ser33/37 Thr41 were significantly increased.Compared with the model group,the cell morphology was significantly improved and the viability was significantly increased,the ATP and lactate content were significantly increased,the expressions of HK1,PKM1 and Wnt3a,pGSK3β Ser9,active β-catenin and nuclear β-catenin protein were significantly upregulat-ed,and the phosphorylation levels of β-catenin Ser33/37 Thr41 were significantly reduced after ICA treatment.However,when the canonical Wnt signaling was inhibited by DKK1,the above effects of ICA on glycolysis were abolished.CONCLUSION ICA exerts neuroprotective effects on Aβ25-35-induced HT22 cell injury by enhancing the glycolysis function through the activation of the Wnt/β-catenin signaling pathway.

5.
Chinese Journal of Behavioral Medicine and Brain Science ; (12): 104-110, 2023.
Article Dans Chinois | WPRIM | ID: wpr-992063

Résumé

Objective:To investigate the icariin on cognitive function and astrocytic pyroptosis in hemorrhagic shock resuscitation model mice.Methods:Forty-eight SPF grade C57BL/6 mice (male) were randomly divided into four groups ( n=12 in each group): Sham operation control group (Group C), hemorrhagic shock and resuscitation group (Group H), hemorrhagic shock and resuscitation plus icariin group (Group HI) and hemorrhagic shock resuscitation plus icariin and SSK1 group (Group HIS, SSK1 was a phosphorylation agonist of mitogen-activated protein kinase p38(p38MAPK). The mice in Group H, HI and HIS were subjected to hemorrhagic shock and resuscitation model by bleeding and retransfusion via left femoral vein; the mice in Group HI and HIS were administered with icariin (10 mg/kg) intragastrically for 7 days; the mice in Group C and H were administered with the same amount of normal saline containing dimethyl sulfoxide(DMSO). The mice in Group HIS were administered with SSK1 (0.5 mg/kg) intraperitoneally, but the mice in Group C, H and HI were only administered with the same amount of normal saline containing DMSO.At 15 days after resuscitation, novel objective recognition test and fear conditioning test were used to assess cognitive dysfunction of mice.Microtubule-associated protein 2(MAP2), a specific marker protein of neurons reflecting astrocytic pyroptosis in the hippocampus of mice, were detected by immunofluorescence assay so as to assess neuronal injury and astrocytic pyroptosis.The levels of IL-1β, IL-18, the ratio of phosphorylated p38MAPK to total p38MAPK in the hippocampus were evaluated by Western blot.SPSS 21.0 software was used for data analysis, multiple samples among groups were compared by one-way ANOVA, and SNK- q test was used for further pairwise comparison. Results:The results of new object recognition test showed that the difference of new object recognition index among the four groups was statistically significant ( F=50.75, P<0.05). The new object recognition indexes in H group(22.7±6.9), HI group(40.1±7.0) and HIS group (22.5±7.5) were significantly lower than that in C group (58.5±11.2). The index in HI group was higher than that in H group, while the index in HIS group was lower than that in HI group (all P<0.05). The results of the fear conditioning test showed that there was a statistically significant difference in the percentage of freezing time among the four groups of mice ( F=60.54, P<0.05). And the percentage of freezing time in H group((21.8±5.0)%), HI group ((38.4±7.4) %)and HIS group((21.3±4.2)%)were lower than that in C group((49.1±7.0)%), which in HI group was higher than that in H group ( P<0.05)and which in HIS group was lower than that in HI group(all P<0.05). The results of immunofluorescence showed that there were significant decreases of MAP2 intensity ((35.3±9.3)%, (63.3±6.1)%, (28.7±10.3)%) but increases of pyroptotic astrocytes ((24.5±4.2)%, (9.3±1.5)%, (22.1±3.3)%) in the H, HI and HIS groups compared with those of C group ((106.7±19.7) %, (3.4±2.0)%). There was an increase of MAP2 intensity but a decrease of pyroptotic astrocytes in the HI group compared with those in H group, and there was a decrease of MAP2 intensity but an increase of pyroptotic astrocytes in the HIS group compared with those of HI group (all P<0.05). The Western blot results showed that there were significant increases of IL-1β, IL-18, the ratio of phosphorylated p38MAPK to total p38MAPK in the H, HI and HIS groups compared with C group, there were decreases of IL-1β, IL-18, the ratio of phosphorylated p38MAPK to total p38MAPK in the HI group compared with H group, and there were increases of IL-1β, IL-18, the ratio of phosphorylated p38MAPK to total p38MAPK in the HIS group compared with those in HI group (all P<0.05). Conclusion:Icariin alleviates hemorrhage shock and resuscitation-induced cognitive dysfunction and astrocytic pyroptosis in mice, and the mechanism may be associated with inhibition of phosphorylated p38MAPK.

6.
China Journal of Chinese Materia Medica ; (24): 6645-6652, 2023.
Article Dans Chinois | WPRIM | ID: wpr-1008862

Résumé

In light of the liver injury risk associated with the oral administration of Xianlin Gubao oral preparation, this study compared the differences in liver injury induced by two different extraction processes in rats and explored the correlation between hepatotoxicity and extraction process from the perspective of the differences in the content of the relevant components. Thirty male Sprague-Dawley(SD) rats were randomly divided into a normal group, tablet extract groups of different doses, and capsule extract groups of different doses, with 6 rats in each group. Each group received continuous oral administration for 4 weeks. The assessment of liver injury caused by different extracts was conducted by examining rat body weight, liver function blood biochemical indicators, liver coefficient, and liver pathological changes. In addition, a high-performance liquid chromatography(HPLC) method was established to simultaneously determine the content of icariin, baohuoside I, and bakuchiol in the extracts to compare the differences in the content of these three components under the two extraction processes. The results showed that both extracts caused liver injury in rats. Compared with the normal group, the tablet extract groups, at the studied dose, led to slow growth in body weight, a significant increase in triglyceride levels(P<0.05), a significant decrease in liver-to-brain ratio(P<0.05), and the appearance of hepatic steatosis. The capsule extract groups, at the studied dose, resulted in slow growth in body weight, a significant increase in aspartate aminotransferase levels(P<0.05), a significant decrease in body weight, liver weight, and liver-to-brain ratio(P<0.05), and the presence of hepatic steatosis and inflammatory cell infiltration. In comparison, the capsule extraction process had a higher risk of liver injury. Furthermore, based on the completion of the liquid chromatography method, the content of icariin and baohuoside Ⅰ in the capsule extract groups was 0.83 and 0.81 times that in the tablet extract groups, respectively, while the bakuchiol content in the capsule extract group was 29.80 times that in the tablet extract groups, suggesting that the higher risk of liver injury associated with the capsule extraction process may be due to its higher bakuchiol content. In summary, the differences in rat liver injury caused by the two extracts are closely related to the extraction process. This should be taken into consideration in the formulation production and clinical application.


Sujets)
Rats , Mâle , Animaux , Rat Sprague-Dawley , Foie/anatomopathologie , Lésions hépatiques dues aux substances/anatomopathologie , Stéatose hépatique , Comprimés , Poids , Extraits de plantes , Phénols
7.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 96-102, 2022.
Article Dans Chinois | WPRIM | ID: wpr-940697

Résumé

ObjectiveTo study the effect of icariin on the proliferative capacity of hepatocellular carcinoma cell line CLC5 and the underlying mechanism. MethodThe targets of icariin were screened out by network pharmacology, and the target network and protein-protein interaction (PPI) network were constructed to predict the possible targets and pathways of icariin. CCK-8 assay was employed to explore the effects of different concentrations (0, 6.25, 12.5, 25, 50 μmol·L-1) of icariin on the viability of CLC5 cells. Further, CLC5 cells were treated with 0, 25, 50 μmol·L-1 icariin, and the effect of icariin on CLC5 cell proliferation was examined by Edu-488 assay and clone formation assay (CFA). Western blot was employed to measure the expression levels of proteins in the protein kinase B (Akt)/glycogen synthase kinase 3β (GSK3β)/cell cycle-dependent kinase (CDK) pathway in the CLC5 cells exposed to different concentrations of icariin. ResultNetwork pharmacological analysis revealed that icariin may inhibit the hepatocellular carcinoma via cell cycle arrest and inhibition of tumor cell proliferation. Compared with the blank group, icariin decreased the viability of CLC5 cells in a time- and concentration-dependent manner (P<0.01) and reduced the positive rate of Edu-488 and the colonies in CFA (P<0.05, P<0.01). Moreover, icariin down-regulated the protein levels of p-Akt, p-GSK3β, CDK4, and CyclinD1 (P<0.05, P<0.01). ConclusionIcariin may block cell cycle to suppress the proliferation of CLC5 cells via inhibiting the Akt/GSK3β/CDK pathway.

8.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 90-97, 2022.
Article Dans Chinois | WPRIM | ID: wpr-943088

Résumé

ObjectiveTo observe the effect of icariin on the recombinant Ras homolog family member A (RhoA)/Rho-associated coiled-coil forming protein kinase (ROCK) signaling pathway in rats with Alzheimer's disease (AD), and to explore the mechanism of icariin in ameliorating the neuronal and dendritic damage. MethodThe β-amyloid 1-42 (Aβ1-42, 2.5 g·L-1) was used to induce AD in rats via lateral ventricle injection, and the rats were divided into a model group, a low-dose icariin group (0.03 g·kg-1), a middle-dose icariin group (0.06 g·kg-1), a high-dose icariin group (0.09 g·kg-1), and a control group. The control group and the model group were given an equal volume of normal saline at a dose of 10 mL·kg-1. The cognitive function of rats was assessed by the Morris water maze. The pathological morphology of the rat hippocampal CA1 area was observed by Nissl staining. Dendritic spine density and dendritic length in the CA1 region of the hippocampus were observed by Golgi-Cox staining. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, RhoA, ROCK1, and ROCK2 in the hippocampus. Western blot assay was used to detect the protein expression levels of TNF-α, IL-1β, IL-6, RhoA, ROCK1, and ROCK2 in the hippocampus. ResultAs compared with the control group, the escape latency of the rats in the model group was increased (P<0.01), while the number of crossing the platform and the dwelling time in the target quadrant were decreased (P<0.01). As compared with the model group, the escape latency of the rats in the middle and high-dose icariin groups was decreased (P<0.05, P<0.01), while the number of crossing the platform and the dwelling time in the target quadrant were increased (P<0.05, P<0.01). As compared with the control group, the number of neurons, dendritic spine density, and dendritic length in the hippocampal CA1 area of the rats in the model group were decreased (P<0.01). As compared with the model group, the number of neurons, dendritic spine density, and dendritic length in the hippocampus of the rats in the middle and high-dose icariin groups were increased (P<0.05, P<0.01). As compared with the control group, the mRNA and protein expression levels of TNF-α, IL-1β, IL-6, RhoA, ROCK1, and ROCK2 in the hippocampus of the rats in the model group were increased (P<0.01). As compared with the model group, the mRNA and protein expression levels of TNF-α, IL-1β, IL-6, RhoA, ROCK1, and ROCK2 in the hippocampus of the rats in the middle and high-dose icariin groups were decreased (P<0.05, P<0.01). ConclusionIcariin improves cognitive function and neuronal and dendritic damage in AD by inhibiting the RhoA/ROCK signaling pathway.

9.
Journal of China Pharmaceutical University ; (6): 215-221, 2022.
Article Dans Chinois | WPRIM | ID: wpr-923498

Résumé

@#To explore the effects and molecular mechanism of icariin on the vascular function of mice with type 1 diabetes induced by alloxan, type 1 diabetic mice model was established by intraperitoneal injection with 200 mg/kg alloxan.After oral administration with icariin (60, 120 mg/kg) daily for 2 weeks, blood glucose, body weight, food intake and water intake were detected.To evaluate the impact of icariin on the function of isolated vascular ring contraction and relaxation, thoracic aortas of mice were removed and the Ach-induced vascular ring relaxation, Phe-induced vascular ring contraction, SNP-induced vascular ring relaxation and KCl-induced vascular ring contraction response were detected.To further confirm the mechanism of icariin to improve vascular function, human umbilical vein endothelial cells (HUVECs) were induced by high glucose (HG) in vitro.Western blot was used to detect the effect of icariin on eNOS, p-eNOS, p38 MAPK and p-p38 MAPK expressions in HG-induced human umbilical vein endothelial cells (HUVECs).The results indicated that icariin significantly ameliorated the weight loss and dampened the increase in water intake of the diabetic mice.Meanwhile, icariin had a certain ameliorative effect on blood glucose and food intake without significant difference.The results of isolated thoracic aortas vascular rings contraction and vasodilation function indicated that icariin significantly improved Phe-induced vascular contraction and Ach‐induced vascular relaxation.Meanwhile, icariin had a certain ameliorative effect on KCl-induced vascular contraction response without significant difference.However, no significant change was observed on endothelium‐independent vascular rings relaxation response induced by SNP after treatment with icariin.Results of Western blot showed that icariin inhibited the expression of p-p38 MAPK and induced expression of p-eNOS in the high glucose-induced HUVECs cell model.Therefore, icariin may attenuate alloxan-induced type 1 diabetic mice vascular diastolic function by inhibiting expression of p-p38 MAPK and inducing expression of p-eNOS.

10.
China Journal of Chinese Materia Medica ; (24): 2170-2177, 2022.
Article Dans Chinois | WPRIM | ID: wpr-928157

Résumé

This study aims to explore the effect of icariin(ICA) on mitochondrial dynamics in a rat model of chronic renal failure(CRF) and to investigate the molecular mechanism of ICA against renal interstitial fibrosis(RIF). CRF was induced in male Sprague-Dawley(SD) rats with 5/6(ablation and infarction, A/I) surgery(right kidney ablation and 2/3 infarction of the left kidney). Four weeks after surgery, the model rats were randomized into the following groups: 5/6(A/I) group, 5/6(A/I)+low-dose ICA group, and 5/6(A/I)+high-dose ICA group. Another 12 rats that received sham operation were randomly classified into 2 groups: sham group and sham+ICAH group. Eight weeks after treatment, the expression of collagen-Ⅰ(Col-Ⅰ), collagen-Ⅲ(Col-Ⅲ), mitochondrial dynamics-related proteins(p-Drp1 S616, p-Drp1 S637, Mfn1, Mfn2), and mitochondrial function-related proteins(TFAM, ATP6) in the remnant kidney tissues was detected by Western blot. The expression of α-smooth muscle actin(α-SMA) was examined by immunohistochemical(IHC) staining. The NRK-52 E cells, a rat proximal renal tubular epithelial cell line, were cultured in vitro and treated with ICA of different concentration. Cell viability was detected by CCK-8 assay. In NRK-52 E cells stimulated with 20 ng·mL~(-1) TGF-β1 for 24 h, the effect of ICA on fibronectin(Fn), connective tissue growth factor(CTGF), p-Drp1 S616, p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 was detected by Western blot, and the ATP content and the mitochondrial morphology were determined. The 20 ng·mL~(-1) TGF-β1-stimulated NRK-52 E cells were treated with or without 5 μmol·L~(-1) ICA+10 μmol·L~(-1) mitochondrial fusion promoter M1(MFP-M1) for 24 h and the expression of fibrosis markers Fn and CTGF was detected by Western blot. Western blot result showed that the levels of Col-Ⅰ, Col-Ⅲ, and p-Drp1 S616 were increased and the levels of p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 were decreased in 5/6(A/I) group compared with those in the sham group. The levels of Col-Ⅰ, Col-Ⅲ, and p-Drp1 S616 were significantly lower and the levels of p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 were significantly higher in ICA groups than that in 5/6(A/I) group. IHC staining demonstrated that for the expression of α-SMA in the renal interstitium was higher in the 5/6(A/I) group than in the sham group and that the expression in the ICA groups was significantly lower than that in the 5/6(A/I) group. Furthermore, the improvement in the fibrosis, mitochondrial dynamics, and mitochondrial function were particularly prominent in rats receiving the high dose of ICA. The in vitro experiment revealed that ICA dose-dependently inhibited the increase of Fn, CTGF, and p-Drp1 S616, increased p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6, elevated ATP content, and improved mitochondrial morphology of NRK-52 E cells stimulated by TGF-β1. ICA combined with MFP-M1 further down-regulated the expression of Fn and CTGF in NRK-52 E cells stimulated by TGF-β1 compared with ICA alone. In conclusion, ICA attenuated RIF of CRF by improving mitochondrial dynamics.


Sujets)
Animaux , Femelle , Humains , Mâle , Rats , Adénosine triphosphate/pharmacologie , Fibrose , Flavonoïdes , Infarctus/anatomopathologie , Rein , Défaillance rénale chronique , Dynamique mitochondriale , Rat Sprague-Dawley , Insuffisance rénale chronique , Facteur de croissance transformant bêta-1/métabolisme
11.
Chinese Pharmacological Bulletin ; (12): 1087-1092, 2021.
Article Dans Chinois | WPRIM | ID: wpr-1014485

Résumé

Abstract; Aim To explore the effect of icariin (ISO) in mice. Methods C57BL/6 mice were ran- (ICA) on myocardial fibrosis induced by isoproterenol domly divided into control group, ISO group, low-dose (15 mg • kg"1), middle-dose (30 mg • kg"1) and high-dose (60 mg • kg"1) of ICA-treated group and Losartan-treated group ( 9 mg • kg"1 ). The control group was subcutaneously injected with normal saline, and the other groups were subcutaneously injected with ISO (5 mg • kg"1, qd) continuously 14 days to established the myocardial fibrosis model. The ICA-treated groups and Losartan-treated group were simultaneously intragastrically administered of ICA or Losartan, respectively. And the other groups received the same a- mount of double distilled water. The left ventricular e- jection fraction (LVEF) and the left ventricular fraction shortening rate ( LVFS) were evaluated by the small animal ultrasound. The heart mass index (HMI) was calculated. The left ventricular collagen deposition was detected by Masson staining. The protein expressions of a-SMA, MMP-2, MMP-9 and TIMP-1 in the left ventricular tissue were detected by Western blot. Results ICA (30, 60 mg • kg"1) and Losartan could inhibit the decreased LVEF and LVFS, the increased HMI and left ventricular collagen deposition, the up- regulated a-SMA and MMP-9 protein expression, the down-regulated MMP-2 and TIMP-1 protein expression in the left ventricular tissues induced by ISO. Conclusions ICA can improve myocardial fibrosis induced by ISO in mice, and the underlying mechanism may be related to the regulation of the protein expression of a- SMA and MMPs/TIMP-1.

12.
China Journal of Chinese Materia Medica ; (24): 5825-5831, 2021.
Article Dans Chinois | WPRIM | ID: wpr-921702

Résumé

This study evaluated the effects of epimedium polysaccharide(EPS) on the solubility of icariin and baohuoside Ⅰ so as to preliminary explore its solubilization function and the underlying mechanism. The solubility of these two insoluble flavonoids in water and polysaccharide solutions was compared by high performance liquid chromatography, and the mechanism was investigated by diffe-rential scanning calorimetry(DSC) and critical micelle concentration determination. The results indicated that their solubilization in crude EPS solutions was concentration-dependent. The solubility of icariin and baohuoside Ⅰ in 20 mg·mL~(-1) EPS-1-1 was 9.05 times and 5.76 times that in water, respectively; while their solubility in 20 mg·mL~(-1) EPS-2-1 was 10.55 and 8.39 times that in water, respectively. The change of the DSC thermograms suggested the formation of new complexes from icariin and baohuoside Ⅰ with polysaccharides. The critical micelle concentrations proved the micellar properties of both EPS-1-1 and EPS-2-1. In short, EPS can significantly increase the solubility of icariin and baohuoside Ⅰ, the mechanism of which may be related to the formation of micellar complexes between EPS and insoluble flavonoids.


Sujets)
Epimedium , Flavonoïdes , Polyosides , Solubilité
13.
China Pharmacy ; (12): 225-230, 2021.
Article Dans Chinois | WPRIM | ID: wpr-862648

Résumé

OBJECTIVE:To establish the method for content determination of 6 components in Fuzheng guben granules ,such as 2,3,5,4′-tetrahydroxystilbene glucoside ,baicalin,icariin,scutellarin,baicalein and wogonin. METHODS :HPLC method was adopted. The determination was performed on Dikma Diamonsil C 18 column with mobile phase consisted of acetonitrile- 0.1% phosphoric acid aqueous solution (gradient elution )at the flow rate of 1.0 mL/min. The detection wavelengths were set at 275 nm (0-8 min),320 nm(8-9 min)and 275 nm(9-33 min). The column temperature was set at 25 ℃,and sample size was 10 μL. With baicalin as reference material ,the relative corr ection factors (fk/s) of other five components were calculated by multi-point correction method and slope correction method ;the retention time difference method was used to locate the chromatographic peaks ; the calculation values obtained by above 2 QAMS were compared with measured values of external standard method. RESULTS : The linear range of 2,3,5,4′-tetrahydroxystilbene glucoside ,baicalin,icariin,scutellarin,baicalein and wogonin were 0.053-2.12, 0.163-6.52,0.059-2.36,0.021 6-0.864,0.03-1.2,0.021-0.84 μg(r>0.999),respectively. RSDs of precision ,stability(12 h)and reproducibility tests were all lower than 3%. Average recoveries were 98.72%-99.82%(RSDs were 0.89%-1.24%,n=9). Using baicalin as reference material ,fk/s of multi-point correction method for 2,3,5,4′-tetrahydroxystilbene glucoside ,icariin,scutellarin, baicalein and wogonin were 1.172,0.528,1.479,1.820 and 2.534,respectively;fk/s of slope correction method were 1.234, 0.550,1.559,1.939,2.664. RSDs of 6 components in 10 batches of Fuzheng guben granules by 3 methods were 0.29%-2.77% (n=10),respectively. Pearson correlation coefficient was not lower than 0.999 9(P<0.001)in measured values between QAMS and external standard method. CONCLUSIONS :QAMS method is established successfully for simultaneous determination of 6 components in Fuzheng guben granules.

14.
Chinese Journal of Tissue Engineering Research ; (53): 2243-2249, 2021.
Article Dans Chinois | WPRIM | ID: wpr-848023

Résumé

BACKGROUND: Icariin is the main effective component of Epimedium, which functions to tonify the kidney, and strengthen tendons and bones. In recent years, a large number of studies have found that icariin plays a significant role in the treatment of osteoarthritis. OBJECTIVE: To review the research progress in the molecular mechanism of icariin in the treatment of osteoarthritis. METHODS: The first author used “Icariin, Osteoarthritis, Cartilage, Subchondral bone, Synovial membrane, synovium, Inflammation" as search words in English and Chinese to search PubMed, CNKI, WanFang, and VIP databases. According to the inclusion criteria and exclusion criteria, 42 articles were included for final analysis. RESULTS AND CONCLUSION: Icariin can promote the cartilage differentiation of bone marrow mesenchymal stem cells and enhance the proliferation of cartilage cells and osteoblasts, to inhibit the degradation of cartilage extracellular matrix, reduce the activity of osteoclasts and alleviate synovial inflammation caused by inflammatory factors. It is an effective treatment for osteoporosis. However, the optimal effective dose and concentration safety of icariin still need a large number of experimental studies. Currently, most of the experiments are still in animal and tissue cell experiments. Numerous clinical studies are needed to continue to explore its specific mechanism in order to provide evidence-based medical evidence for icariin in the treatment of osteoarthritis.

15.
China Journal of Chinese Materia Medica ; (24): 4497-4503, 2021.
Article Dans Chinois | WPRIM | ID: wpr-888151

Résumé

This study aimed to observe the inhibitory effect of icariin against oxidative stress-induced calcification in aortic vascular smooth muscle cells(VSMCs) and elucidate the molecular mechanism of icariin in inhibiting endoplasmic reticulum stress(ERS)-mediated atherosclerotic calcification, so as to provide new ideas for exploring the anti-atherosclerotic mechanism of Epimedii Folium. The VSMCs in rat thoracic aorta were subjected to adherent culture and then treated with the complete calcification DMEM containing high glucose and hydrogen peroxide(H_2O_2) for three weeks. The resulting calcified VSMCs were divided into different treatment groups. Icariin was added one week after calcification induction for protecting the VSMCs, whose viability was then detected using cell counting kit-8(CCK-8). Alizarin red-S staining was conducted to observe the calcification degree. The activity of alkaline phosphatase(ALP) in VSMCs was measured using the disodium phenyl phosphate substrate and the calcium content was measured by arsenazo Ⅲ method. The mRNA expression levels of ossification-related factors including osteocalcin(OC), osteopontin(OPN), Runt-related transcription factor 2(Runx2), and type Ⅰ collagen(Col Ⅰa) were detected by real-time PCR. Western blot was carried out to determine the protein expression levels of α-smooth muscle actin(α-SMA), Runx2, activating transcription factor 4(ATF4), and eukaryotic translation initiation factor(eIF)-2α. The results showed that H_2O_2 significantly induced the calcification of VSMCs, increased the ALP activity and calcium content in VSMCs, promoted OC, OPN, Runx2, and Col Ⅰa mRNA expression and Runx2 protein expression, and reduced α-SMA protein expression. The ATF4 protein expression and eIF2α phosphorylation were also elevated significantly. Icariin reversed the calcification of VSMCs induced by H_2O_2, inhibited ALP activity and calcium content in VSMCs, down-regulated the mRNA expression levels of OC, OPN, Runx2 and Col Ⅰa and Runx2 protein expression, and relatively up-regulated the expression of α-SMA. The expression of ATF4 and phosphorylation of eIF2α also declined significantly. All these have demonstrated that icariin inhibited VSMCs calcification by down-regulating the ossification-related factors and lowering ALP activity and calcium content in VSMCs. Besides, the down-regulation of Runx2 expression and the inhibition of ATF4 and eIF2α-mediated cellular calcification pathway in ERS might also be involved in such calcification-suppressing process.


Sujets)
Animaux , Rats , Cellules cultivées , Flavonoïdes/pharmacologie , Muscles lisses vasculaires/métabolisme , Myocytes du muscle lisse , Stress oxydatif
16.
China Pharmacy ; (12): 812-818, 2021.
Article Dans Chinois | WPRIM | ID: wpr-875813

Résumé

OBJECTIVE:To study the improvement effects of ica riin(ICA)on cognitive function in schizophrenia model rats and its mechanism. METHODS :SD rats were divided into blank control group ,model group ,ICA low-dose ,medium-dose and high-dose groups (15,30,60 mg/kg). Except for blank control group ,other groups were given N-methyl-D-aspartate receptor antagonist MK- 801(0.2 mg/kg)intraperitoneally to induce schizophrenia rats models ,once a day ,for consecutive 14 days. After modeling,ICA groups were intragastrically administered with the corresponding drugs ,while blank control group and model group were intragastrically administered with the same volume of water ,once a day ,for consecutive 7 days. The behavioral com changes of rats were detected by Morris water maze test ,open field test , forced swimming test and Y maze test pathological changes of hippocampus were observed by Nissl staining;the levels of cholinergic indexes [acetylcholine (Ach),choline acetyltransferase (ChAT) and acetylcholinesterase (AchE)] in cerebral tissues were detected by ELISA. The expression of BDNF ,ERK and CREB mRNA in cerebral tissue were detected by RT-PCR ;expression or phosphorylation level of BDNF ,ERK,CREB protein ,apoptosis related proteins (Bcl-2,Bax and Caspase- 3)were detected by Western blot. RESULTS :Compared with blank control group ,escape latency ,distance at T 1-T3, cumulative immobility time and the expression of Caspase- 3 protein in cerebral tissues were significantly increased in model group (P<0.05);the times of crossing platform ,alternation rate ,the number of Nissl staining positive neurons in hippocampus tissues , the levels of Ach and ChAT in cerebral tissues ,Bcl-2/Bax ratio ,mRNA and protein expression of BDNF ,mRNA expression of ERK and CREB ,the phosphorylation of ERK 1/2 and CREB were significantly decreased (P<0.05).Compared with model group , escape latency ,distance at T 1-T3,cumulative immobility time ,the number of Nissl staining positive neurons ,AchE level in cerebral tissues and relative expression of Caspase- 3 protein were significantly decreased in ICA high-dose group (P<0.05);the times of crossing platform ,alternation rate ,levels of Ach and ChAT in cerebral tissues ,Bcl-2/Bax ratio ,mRNA and protein expression of BDNF ,mRNA expression of ERK and CREB ,the phosphorylation of ERK 1/2 and CREB were increased significantly(P<0.05). Above indexes in ICA low-dose and medium-dose groups were partially improved significantly than model group(P<0.05). CONCLUSIONS :ICA can improve cognitive function in schizophrenia model rats.Its mechanism may be related to regulating cholinergic system ,inhibiting neuronal apoptosis ,and promoting the expression of BDNF/ERK/CREB signaling pathway.

17.
Chinese Journal of Tissue Engineering Research ; (53): 1988-1992, 2020.
Article Dans Chinois | WPRIM | ID: wpr-847674

Résumé

BACKGROUND: Iron accumulation or iron overload is closely related to the proliferation and apoptosis of bone marrow mesenchymal stem cells, which may play an important role in the pathological process of cardiovascular diseases. OBJECTIVE: To investigate the regulatory effect of icariin on ferroptosis of bone marrow mesenchymal stem cells and its differentiation into cardiomyocytes. METHODS: Taking bone marrow mesenchymal stem cells as the research object, the blank group was cultured for 14 days. The model group was treated with 0.4 μmol/L Erastin for 14 days to establish ferroptosis model. The treatment group was treated with 80 nmol/L icariin and 0.4 μmol/L Erastin for 14 days. CCK-8 method was used to detect the proliferation of cells in each group, and the levels of malondialdehyde and superoxide dismutase were detected by related kits. Western blot assay was used to detect the expression of TFR1, FPN1, Bcl-2 and Bax protein. RT-PCR was used to detect the mRNA expression of GATA4 and cTnT. RESULTS AND CONCLUSION: (1) Compared with the model group, the cell proliferation ability of the treatment group was significantly improved (P < 0.05). (2) Compared with the model group, the level of malondialdehyde in the lysate of bone marrow mesenchymal stem cells in the treatment group was significantly lower (P < 0.05), while the level of superoxide dismutase was significantly increased (P < 0.05). (3) Compared with the model group, TFR1 protein expression was decreased (P < 0.05), FPN1 protein expression was increased (P < 0.05), and Bcl-2/Bax protein expression ratio was increased significantly (P < 0.05) in the treatment group. (4) Compared with the model group, the expression of GATA4 and cTnT mRNA was significantly increased in the treatment group (P < 0.05). (5) The results showed that icariin could inhibit the ferroptosis of bone marrow mesenchymal stem cells and improve the ability of bone marrow mesenchymal stem cells to differentiate into cardiomyocyte-like cells.

18.
Chinese Journal of Tissue Engineering Research ; (53): 4218-4223, 2020.
Article Dans Chinois | WPRIM | ID: wpr-847356

Résumé

BACKGROUND: Nonunion is a common clinical complication in orthopedics, which seriously impacts the physical and mental health and quality of life of patients. In recent years, a large number of studies have found that icariin plays a significant role in promoting fracture healing and treating bone defects. Bone nonunion and fracture healing coexist, and the research on the mechanism of fracture healing actually focuses on the treatment of bone nonunion. OBJECTIVE: To review the research progress in the molecular mechanism of icariin in the treatment of bone nonunion. METHODS: The first author used “icariin, bone nonunion, bone marrow mesenchymal stem cells, periosteal cell, osteoblasts, osteoclast” as key words in English and Chinese to search PubMed, CNKI, WanFang and VIP databases. A total of 542 articles were retrieved and screened manually according to the selection criteria and exclusion criteria. Finally, 44 articles were included for result analysis. RESULTS AND CONCLUSION: Icariin can effectively promote fracture healing and treat bone nonunion by promoting the proliferation and differentiation of bone marrow mesenchymal stem cells and periosteal cells, promoting the proliferation and maturation of osteoblasts and inhibiting the osteoclast effect of osteoclasts. However, most of the experiments are still in the basic experimental research, and there is still a need for a large number of clinical studies as well as studies on related proteins and genes, to provide a new idea for the clinical use of Chinese herbs in the treatment of bone nonunion.

19.
Chinese Journal of Tissue Engineering Research ; (53): 3974-3980, 2020.
Article Dans Chinois | WPRIM | ID: wpr-847318

Résumé

BACKGROUND: Bone tissue engineering has provided a novel ideal for treating bone defects in clinic. This study is the first to combine traditional Chinese medicine with the nanostructures of tissue-engineered scaffolds in order to explore and construct a new bone tissue substitute material for the treatment of bone defects. OBJECTIVE: To investigate the osteogenic activity of icariin (ICA)/hydroxyapatite (HA)/poly(lactic-co-glycolic acid) (PLGA) composite scaffolds. METHODS: A HA/PLGA composite scaffold was prepared by physical blending of HA and PLGA, and was then soaked in ICA solution of different concentrations to obtain the HA/ICA/PLGA scaffold. Rabbit bone marrow mesenchymal stem cells were used to evaluate the cell adhesion, proliferation, osteogenesis and cytotoxicity of the composite scaffold. The cell adhesion, proliferation and cytotoxicity were detected by MTT method. The activities of alkaline phosphatase and osteocalcin were detected by ELISA. The expression levels of osteogenic genes and proteins were detected by fluorescence quantitative PCR and western blot assay, respectively. RESULTS AND CONCLUSION: Adding appropriate amount of HA into PLGA could improve the mechanical strength of the scaffold, and 10% HA had the best effect with tensile strength of (1.67±0.37) MPa, and compression modulus of (4.17±1.62) MPa, and nanostructure would be formed on the surface of the scaffold. The nanostructure could promote the adhesion of bone marrow mesenchymal stem cells on the surface of the scaffold. ICA did not affect the proliferation of bone marrow mesenchymal stem cells on the composite scaffold. However, the HA/PLGA composite scaffold soaked in 1.00 µmol/L ICA aqueous solution had the optimal osteogenic differentiation function, and the expression levels of alkaline phosphatase, osteocalcin, osteogenic related genes and proteins (Runx-2 and COL I) were increased. The ICA/HA/PLGA scaffold had no cytotoxicity. These results suggest that HA (10%)/ICA (1.00 µmol/L)/PLGA scaffold has good mechanical properties, osteogenesis and biocompatibility, which has the potential to be a favorable scaffold for bone tissue engineering.

20.
Chinese Traditional and Herbal Drugs ; (24): 190-196, 2020.
Article Dans Chinois | WPRIM | ID: wpr-846707

Résumé

Objective: To compare the flavonoids contained in different parts of different botanical origins of Epimedii Folium, and provide a basis for the quality evaluation of Epimedii Folium and the reasonable selection of medicinal parts. Methods: The aerial parts of 13 batches of Epimedii Folium were collected and divided into three parts: leaf, petiole and stem. The HPLC fingerprint and content of five flavonoids, including epimedin A, epimedin B, epimedin C, icariin and baohuoside I, were analyzed. Then the analysis of variance and the similarity evaluation software of traditional Chinese medicine chromatographic fingerprint were used. Combined with cluster analysis (HCA), the content differences of flavonoids in leaf, petiole and stem of Epimedii Folium were evaluated. Results: The fingerprints showed that the chemical constituents in Epimedii Folium leaves were richer than stems and petioles, and the chemical constituents in petioles and stems were basically the same. The content of five components in leaves was significantly higher than that of petiole and stem, even up to 10 times. Cluster analysis also showed that the leaves were obviously distinct from the petiole and stem. Conclusion: The quality differences of Epimedii Folium leaves, petioles and stems were clarified, and this study can provide the scientific evidence for the selection of medicinal parts and quality control of Epimedii Folium.

SÉLECTION CITATIONS
Détails de la recherche