Objective To investigate the intervention effect of Xiao'er Zhixiao Pingchuan Granules(XEZXPCG)on ovalbumin(OVA)-induced airway remodeling in asthmatic mice and its potential mechanism by regulating macrophage migration inhibitory factor(MIF)and inhibiting the phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)signaling pathway.Methods A total of 96 SPF-grade male BALB/c mice were randomly divided into blank,model,XEZXPCG low/medium/high-dose groups(2.05,4.10,and 8.20 g/kg),adeno-associated virus(AAV)NC shRNA,AAV MIF shRNA(MIF gene silencing),and LY294002(PI3K/Akt inhibitor,1 mg/kg)groups(12 mice in each group).Asthma models were established through OVA sensitization and challenge.Airway resistance and the proportions of inflammatory cells(eosinophils and macrophages)in bronchoalveolar lavage fluid(BALF)were detected.Serum inflammatory factor(OVA-IgE,interleukin[IL]-1β,IL-6,tumor necrosis factor-alpha,and interferon-gamma)levels and BALF were quantified.Hematoxylin and eosin,Masson,and periodic acid-Schiff staining were used to evaluate airway wall thickness(Wat/Phm),smooth muscle area(Wam/Phm),collagen deposition,and goblet cell metaplasia.Western blotting,immunofluorescence,and real-time fluorescence-qPCR were used to detect MIF protein and mRNA expressions,as well as activation markers of the PI3K/Akt pathway and cell cycle-related proteins(including cyclin-dependant kinase 6[CDK6],Cyclin D1,Cyclin D3,and p21),in lung tissues.Results Compared to the model group,a XEZXPCG medium or high-dose significantly reduced airway resistance(P<0.05),improved the imbalance of eosinophil and macrophage proportions in BALF,and decreased inflammatory factor levels in serum and BALF(P<0.05).XEZXPCG medium or high-dose alleviated airway epithelial damage,goblet cell hyperplasia,and collagen fiber deposition,and reduced the Wat/Phm and Wam/Phm(P<0.05),with effects comparable to those of the AAV MIF shRNA and LY294002 groups.XEZXPCG medium and high-inhibited MIF protein/mRNA expression(P<0.05),downregulated Akt phosphorylation(P<0.05),upregulated p21 protein expression,and downregulated Cyclin D1,Cyclin D3,and CDK6 expressions(P<0.05).Conclusion XEZXPCG alleviates airway inflammation and improves airway remodeling in OVA-induced asthmatic mice by inhibiting MIF expression,downregulating the PI3K/Akt signaling pathway,and regulating cell cycle progression.XEZXPCG enhances airway remodeling through MIF-mediated PI3K/Akt pathway regulation.

Objective To investigate the intervention effect of Xiao'er Zhixiao Pingchuan Granules(XEZXPCG)on ovalbumin(OVA)-induced airway remodeling in asthmatic mice and its potential mechanism by regulating macrophage migration inhibitory factor(MIF)and inhibiting the phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)signaling pathway.Methods A total of 96 SPF-grade male BALB/c mice were randomly divided into blank,model,XEZXPCG low/medium/high-dose groups(2.05,4.10,and 8.20 g/kg),adeno-associated virus(AAV)NC shRNA,AAV MIF shRNA(MIF gene silencing),and LY294002(PI3K/Akt inhibitor,1 mg/kg)groups(12 mice in each group).Asthma models were established through OVA sensitization and challenge.Airway resistance and the proportions of inflammatory cells(eosinophils and macrophages)in bronchoalveolar lavage fluid(BALF)were detected.Serum inflammatory factor(OVA-IgE,interleukin[IL]-1β,IL-6,tumor necrosis factor-alpha,and interferon-gamma)levels and BALF were quantified.Hematoxylin and eosin,Masson,and periodic acid-Schiff staining were used to evaluate airway wall thickness(Wat/Phm),smooth muscle area(Wam/Phm),collagen deposition,and goblet cell metaplasia.Western blotting,immunofluorescence,and real-time fluorescence-qPCR were used to detect MIF protein and mRNA expressions,as well as activation markers of the PI3K/Akt pathway and cell cycle-related proteins(including cyclin-dependant kinase 6[CDK6],Cyclin D1,Cyclin D3,and p21),in lung tissues.Results Compared to the model group,a XEZXPCG medium or high-dose significantly reduced airway resistance(P<0.05),improved the imbalance of eosinophil and macrophage proportions in BALF,and decreased inflammatory factor levels in serum and BALF(P<0.05).XEZXPCG medium or high-dose alleviated airway epithelial damage,goblet cell hyperplasia,and collagen fiber deposition,and reduced the Wat/Phm and Wam/Phm(P<0.05),with effects comparable to those of the AAV MIF shRNA and LY294002 groups.XEZXPCG medium and high-inhibited MIF protein/mRNA expression(P<0.05),downregulated Akt phosphorylation(P<0.05),upregulated p21 protein expression,and downregulated Cyclin D1,Cyclin D3,and CDK6 expressions(P<0.05).Conclusion XEZXPCG alleviates airway inflammation and improves airway remodeling in OVA-induced asthmatic mice by inhibiting MIF expression,downregulating the PI3K/Akt signaling pathway,and regulating cell cycle progression.XEZXPCG enhances airway remodeling through MIF-mediated PI3K/Akt pathway regulation.

Humans ; Artificial Intelligence ; Algorithms ; Brain ; Head

Objective To explore the potential mechanism underlying the treatment of pediatric asthma using Xiaoer Zhixiao Pingchuan Granules through network pharmacology analysis and animal experimental validation.Methods Active components and their associated targets in Xiaoer Zhixiao Pingchuan Granules were identified through screening and retrieval of TCMSP,BATMAN-TCM,and UniProt databases.Disease-related targets for pediatric asthma were selected from GeneCards,DisGeNET,and OMIM databases.The target protein-protein interaction(PPI)relationship between the intersecting targets of the two was obtained through the STRING database,and import it into Cytoscape 3.8.0 software to construct a PPI network.GO and KEGG enrichment analyses were conducted using the Metascape platform to identify potential pathways.An asthmatic mouse model was induced by ovalbumin,and different concentrations of Xiaoer Zhixiao Pingchuan Granules were administered as interventions.Histopathological changes were evaluated using HE staining and PAS staining,and the network pharmacology findings were validated through Western blot analysis.Results A total of 154 active ingredients targeting 283 pediatric asthma-related genes were identified in Xiaoer Zhixiao Pingchuan Granules.KEGG enrichment analyses revealed significant enrichment of signaling pathways such as the PI3K-Akt signaling pathway,TNF signaling pathway,and MAPK signaling pathway among intersection targets.Thirteen key targets were identified through topological analysis of ingredients-targets-pathways network.Animal experiments demonstrated that Xiaoer Zhixiao Pingchuan Granules significantly alleviated ovalbumin-induced airway inflammation and goblet cell hyperplasia,while downregulating the expression of key proteins in the PI3K-Akt signaling pathway(P<0.05).Conclusion The therapeutic efficacy of Xiaoer Zhixiao Pingchuan Granules in pediatric asthma involves a multi-pathway and multi-target mechanism,with the PI3K-Akt signaling pathway emerging as a potential key molecular target.

Objective:To study the influence of rutin in the morphology of renal tissue of the diabetic mice induced by streptozotocin(STZ), and to clarify the effect of rutin on the kidney tissue damage.Methods:Twelve mice of the total 70 Kunming mice were used as normal group, the other were used to estabish type 1 diabetes mouse models by intraperitoneally injected with STZ (62.5 mg·kg-1), once daily for 5 d.The successfully established model mice were randomly divided into model group,low dose (50 mg·kg-1)of rutin group, high dose (100 mg·kg-1) of rutin group and irbesartan group (45 mg·kg-1).The mice in model group and normal group were given the carboxy methyl cellulose(CMC) and the other mice were given drugs by intragastric administration once daily for 8 weeks accordingly.The weight and blood glucose of the each mouse were determined.Full automatic biochemical analyzer was used to detect the levels of serum creatinine (Cre) and blood urine nitrogen (BUN) of the mice , and the kidney index was calculated.The morphology of renal tissue was observed by HE staining, Masson staining and electron microscope.Results:After injection of STZ,the model success rate was up to 98%.Compared with normal group, there was no significant difference in the weight of the mice in other groups before administration(P>0.05).After administration of rutin, the weights of the mice in model group, low dose of rutin group, high dose of rutin group and irbesartan group were significantly decreased (P<0.05).Compared with model group, the levels of blood glucose of the mice in low and high doses of rutin groups were significantly decreased(P<0.05);the levels of Cre and BUN were significantly reduced (P<0.05);the kidney index of the mice in high dose of rutin group was significantly decreased (P<0.05).Compared with normal group,the kidney tissue of the mice in model group was seriously damaged;glomerular was weaked, the kidney tissue fibrosis was serious, glomerular basement membrane was diffusely thickened and foot process was coalesced or overgrow.Compared with model group,the degree of injury of the mice in low and high doses of rutin groups were significantly improved, especially in high dose of rutin group.Conclusion:Rutin can improve the renal function of diabetic mice induced by STZ and reduce the degree of renal tissue damage in the diabetic mice

Objective To investigate the expression of parvalbumin (PV)in the cognitive im-pairment mice induced by sevoflurane anesthesia,and to explore whether enriched environment could reverse it.Methods One hundred and forty-four Six-day-old C57BL/6 male mice at postnatal day 6 were randomly divided into the following four groups (n =36):control+standard environment group (group CS),control+enriched environment group(group CE),sevoflurane anesthesia+standard en-vironment group(group SS)and sevoflurane anesthesia+enriched environment group(group SE).An-imals were exposed to 3% sevoflurane plus 30% O 2 or 30% O 2 2 h daily for 3 days from postnatal day 6 (P6)to P8.The exposed pups were randomly allocated to an enriched environment for 2 h daily between P8 and P90 or to a standard environment.Western blotting were used for determining PV ex-pression in the prefrontal cortex and hippocampus at P9,P14,P42,P60 and P90.Cognitive functions were assessed by performing the open field (P41 ),and fear conditioning tests (P42-43 and P89-90, respectively).Results In the open field test,there was no significant difference in the total travel dis-tance and the time spent in the center of the arena among groups.In the contextual fear condition test, compared with group CS,the group SS only exhibited a reduced freezing response in the contextual test at P43,but not at P90.In the cued fear conditioning test,no difference was observed in the freez-ing time among the four groups.The PV expression in the prefrontal cortex and hippocampus of group SS were significantly lower than that of group CS at P9 and P14 (P <0.001),while recovered at P60 and P90.The PV expression in the prefrontal cortex and hippocampus of group SE were significantly higher than that of group SS at P42 (P <0.05).Linear regression analysis demonstrated a positive correlation between the PV expression in the prefrontal cortex and the freezing time induced by envi-ronment (r =0.670 7,P =0.000 1),and there was a positive correlation between the PV expression in hippocampus and the freezing time induced by environment (r = 0.509 6,P = 0.001 9 ). Conclusion The cognitive impairment induced by sevoflurane anesthesia may be associated with the reduction of PV in the prefrontal cortex and hippocampus.Placement of the sevoflurane-exposed mice in an enriched environment prevented the development of these abnormalities.

AIM:To investigate the effect of signal transducer and activator of transcription 1 ( STAT 1 ) on proliferation and interferon-β(IFN-β) sensitivity of human non-small-cell lung cancer H1299 cells.METHODS:STAT1 or EGFP gene was transfected into H1299 cells by the lentiviral vectors system.The cell number was counted under a mi-croscope and cell proliferation was tested by MTT assay.In addition, the cells transfected with STAT1 and EGFP were trea-ted with IFN-βand cell viability was measured by MTT assay.The protein levels of p-STAT1, ICAM-1 and PCNA were de-tected by Western blot.RESULTS: Over-expression of STAT1 inhibited H1299 cell proliferation (P<0.05).H1299 cells transfected with STAT1 gene had a higher sensitivity to IFN-βthan the control cells transfected with EGFP ( P <0.05).Overexpression of STAT1 increased the protein level of p-STAT1, and reduced IACM-1 expression in H1299 cells. Moreover, STAT1 enhanced STAT1 phosphorylation and downregulated the expression of PCNA in H1299 cells treated with IFN-β.CONCLUSION:STAT1 inhibits the proliferation and enhances the IFN-βsensitivity of non-small-cell lung cancer H1299 cells.