ObjectiveTo investigate the effect of mesenchymal stem cells (MSCs) transfected with hepatic growth factor (HGF) gene on the survival volume of free grain fat grafts.Methods MSCs of male Wistar rats were transfected with Ad-GFP or Ad-HGF.The transfection infectivity of Ad-GFP to MSCs,the expression of HGF were measured using ELISA assay.150 rats were randomly divided into 5 groups:control group (group A),MSCs group (group B),Ad HGF group (group C),MSCs transplanted with Ad-GFP group (group D),and MSCs transplanted with Ad-HGF (MSCsHGF) group (group E).Then,the same volume of frain fat graft,mixed with DMEM LG andMSCs,Ad HGF,MSCs-GFP,MSCs HGF respectively,were transplanted to rats back,but control group were only mixed with DMEM-LG.Fat graft was obtained on days 3,5,7,14,28,and 60 after implantation.The volume of fat graft was measured by messcylinder,and the expression of HGF and CD34 in transplanted fat tissue were detected by immunohistochemistry.ResultsThe transfection infectivity of Ad-GFP to MSCs was 89.6 ％ at 100 MOI,the expression of HGF in MSCs culture medium reached to the level after being transfected with Ad-HGF for 48 h.Compared with other 4 groups,at days 3,5,7,and 14 post-transplantation,the expression of HGF in E group transplanted fat of group E had statistics significance (P＜0.05).The persentage of survival volume of fat graft in group E was significantly higher than that of other group ( P＜0.05) at days 28,and 60 post transplantation.ConclusionsMSCs transplanted with Ad-HGF could secrete HGF and increase the survival volume of fat grafts.

OBJECTIVE: To investigate 18β-sodium glycyrrhetinic acid impact on nasal mucosa epithelial cilia in rat models of allergic rhinitis (AR). METHOD: AR models were established by ovalbumin-induction. Wister rats were randomly divided into groups as normal group, model group, budesonide (0.2 mg/kg) group and sodium glycyrrhetinic acid (20 mg/kg and 40 mg/kg) group after the success of AR models. At 2 weeks and 4 weeks after treatment, the behavioral changes of rats were observed and recorded, and nasal septum mucosae were collected after 2 week and 4 week intervention, and the morphological changes of nasal mucosae were observed by electron microscope. RESULT: Model group developed typical AR symptoms, the total score in all animals was > 5. With budesonide and sodium glycyrrhetinic acid treatment, the AR symptoms were relieved, and the total scores were reduced significantly (P < 0.01). Compared with the model group: after 2 weeks' intervention, thick mucous secretions on the top of columnar epithelium cilia in rat nasal mucosa was significantly reduced, and cilia adhesion, lodging, shedding were relieved in budesonide group and sodium glycyrrhetinic acid group, the relieve in budesonide group was slightly better than that in sodium glycyrrhetinic acid group; after 4 week intervention, Cilia adhesion, lodging, shedding were completely vanished, and the cilia were ranged in regular direction in budesonide group and sodium glycyrrhetinic acid group. Cilia in sodium glycyrrhetinic acid (20 mg/kg) group was more orderly, smooth than that in budesonide group and sodium glycyrrhetinic acid group (40 mg/kg), and the condition of cilia in sodium glycyrrhetinic acid group (20 mg/kg) was similar to the normal group. CONCLUSION 18β-sodium glycyrrhetinic acid is effective to restrain the pathological changes of nasal mucosa cilia in rat models of AR.
Animals ; Budesonide ; pharmacology ; Cilia ; drug effects ; Disease Models, Animal ; Glycyrrhetinic Acid ; analogs & derivatives ; pharmacology ; Nasal Mucosa ; drug effects ; Ovalbumin ; Random Allocation ; Rats ; Rhinitis, Allergic ; drug therapy

OBJECTIVE: To observe 18β-glycyrrhetinic acid (GA) impact on ultrastructure of tight junctions (TJs) of nasal mucosa epithelial cells in rats models of allergic rhinitis (AR). METHOD: Ninety-six Wistar rats were randomly divided into control group, model group, loratadine group, and 18β-glycyrrhetinic acid group, and each group had 24 rats. Ovalbumin was used to establish a rat AR model. The behavioral changes and the tight junctions of nasal epithelial were observed and compared in different groups after 2,4,6 and 10 weeks intervention. RESULT: The length of TJs in allergic rhinitis model became shorter, electron-high-density plasma membrane became thicker, number of the integration loci reduced and gap of TJs widened or even ruptured. With the consistent effect of allergens,the changes of TJs in the model group aggravated gradually,and the changes of ultrastructure of TJs in 18β-glycyrrhetinic acid group was relieved apparently compared to model group and even were close to the control model with time. CONCLUSION 18β-glycyrrhetinic acid can recover the ultrastructure of the tight junctions of AR rat nasal epithelial cells.
Animals ; Cell Count ; Epithelial Cells ; ultrastructure ; Glycyrrhetinic Acid ; analogs & derivatives ; pharmacology ; Nasal Mucosa ; cytology ; Ovalbumin ; Rats ; Rats, Wistar ; Rhinitis, Allergic ; drug therapy ; Tight Junctions ; drug effects

Objective: To investigate the effect of methyleugenol on expression of MUC5AC in nasal mucosa of rats with allergic rhinitis (AR).Methods: Seventy-two Wistar rats were randomly divided into 6 groups: normal control group , AR group , loratadine group , low-dose methyleugenol group , middle-dose methyleugenol group and high-dose methyleugenol group with 12 rats in each group . AR was induced by intraperitoneal injection of ovalbumin in latter 5 groups .10 mg loratadine q .d was given to rats in loratadine group by gavage; and 10 mg/kg, 20 mg/kg and 40 mg/kg methyleugenol were given by gavege q .d to rats in low-, middle-and high-dose methyleugenol groups , respectively .Nasal mucosa samples were obtained from rats at 1, 2, 4 and 6 weeks after drug intervention .The expression of MUC5AC protein and mRNA in nasal mucosa was detected by immunohistochemistry and real-time fluorescence quota PCR ( RT-PCR ) , respectively .Results: Compared with AR , the percentage of cells staining positively for MUC 5AC protein and the relative quantity of MUC5AC mRNA in middle-and high-dose methyleugenol groups were significantly decreased after 2 and 4 weeks of drug intervention ( P<0.05 ) , but no such decrease was observed in low-dose methyleugenol group at all time points ( P >0.05 ) .The percentage of cells with positive expression of MUC 5 AC protein and mRNA in loratadine group were significantly decreased after 1 week of administration ( P <0.05 ) .The percentage of cells with positive MUC 5 AC protein in middle-dose methyleugenol group was higher than that in loratadine group ( P<0.05 ) after 6 week of drug intervention , but the difference was not seen in high-dose group ( P >0.05 ) . There was no significant difference in relative quantities of MUC 5 AC mRNA after 4 weeks of administration between high-and middle-dose methyeugenol groups and loratadine group ( P >0.05 ) . Conclusion:Methyleugenol can attenuate AR through inhibiting the expression of MUC5AC mRNA and protein in nasal mucosa of AR rats .

To investigate the effect of methyleugenol on expression of MUC5AC in nasal mucosa of rats with allergic rhinitis (AR).Seventy-two Wistar rats were randomly divided into 6 groups:normal control group, AR group, loratadine group, low-dose methyleugenol group, middle-dose methyleugenol group and high-dose methyleugenol group with 12 rats in each group. AR was induced by intraperitoneal injection of ovalbumin in latter 5 groups. 10 mg loratadine q.d was given to rats in loratadine group by gavage; and 10 mg/kg, 20 mg/kg and 40 mg/kg methyleugenol were given by gavege q.d to rats in low-, middle-and high-dose methyleugenol groups, respectively. Nasal mucosa samples were obtained from rats at 1, 2, 4 and 6 weeks after drug intervention. The expression of MUC5AC protein and mRNA in nasal mucosa was detected by immunohistochemistry and real-time fluorescence quota PCR (RT-PCR), respectively.Compared with AR, the percentage of cells staining positively for MUC5AC protein and the relative quantity of MUC5AC mRNA in middle-and high-dose methyleugenol groups were significantly decreased after 2 and 4 weeks of drug intervention (<0.05), but no such decrease was observed in low-dose methyleugenol group at all time points (>0.05). The percentage of cells with positive expression of MUC5AC protein and mRNA in loratadine group were significantly decreased after 1 week of administration (<0.05). The percentage of cells with positive MUC5AC protein in middle-dose methyleugenol group was higher than that in loratadine group (<0.05) after 6 week of drug intervention, but the difference was not seen in high-dose group (>0.05). There was no significant difference in relative quantities of MUC5AC mRNA after 4 weeks of administration between high-and middle-dose methyeugenol groups and loratadine group (>0.05).Methyleugenol can attenuate AR through inhibiting the expression of MUC5AC mRNA and protein in nasal mucosa of AR rats.
Animals ; Dose-Response Relationship, Drug ; Down-Regulation ; drug effects ; Eugenol ; analogs & derivatives ; pharmacology ; Loratadine ; Mucin 5AC ; drug effects ; physiology ; Nasal Mucosa ; chemistry ; Ovalbumin ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Rhinitis, Allergic ; chemically induced ; drug therapy ; physiopathology

Objective: To explore the effect of 18β-sodium glycyrrhetinic acid on thymic stromal lymphopoietin (TSLP) in nasal mucosa of allergic rhinitis (AR) rats. Methods: One hundred Wistar rats,half male and half female,were randomly divided into 5 groups by random number table method: control group, AR model group,budesonide group,18β-sodium glycyrrhetinic acid at dose of 20 mg/kg and 40 mg/kg groups, with 20 rats in each group. AR animal models were established by ovalbumin (OVA) sensitization in the other four experimental groups. After successful modeling, budesonide and 18β-sodium glycyrrhetinic acid were given in each group,and the detection time points were 2 weeks and 4 weeks. The distribution of TSLP in rat nasal mucosa was detected by immunohistochemistry,and the expression of TSLP in rat nasal mucosa was determined by Western blot at the protein level. The expression of TSLP-mRNA in rat nasal mucosa was detected and compared by real-time fluorescence quantitative PCR (RT-PCR) at mRNA level. The concentrations of IL-4 and OVA-sIgE in rat serum were measured and compared by ELISA. One-way analysis of variance and the least significant difference method were used for the comparison among groups, LSD t test was used for the comparison between the two groups,and the difference was statistically significant (P<0.05). Results: Immunohistochemistry confirmed existence of TSLP in rat nasal mucosa, especially in epithelial cells,endothelial cells and epithelial cilia. Western blot and RT-PCR suggested that the expression of TSLP and TSLP-mRNA in nasal mucosa of AR model group was significantly higher than that of control group (2 weeks TSLP: 1.795 9±0.131 4 vs 0.990 5±0.164 2, 4 weeks TSLP: 1.809 7±0.253 4 vs 0.870 3±0.124 4; 2 weeks TSLP-mRNA:4.582 9±0.697 7 vs 1.108 7±0.081 1, 4 weeks TSLP-mRNA:4.814 4±0.662 8 vs 1.001 0±0.155 3; all P<0.05). After 2 weeks and 4 weeks of drug intervention,the expression of TSLP and TSLP-mRNA was inhibited in nasal mucosa of budesonide group,18β-sodium sodium glycyrrhetinic acid at dose of 20 mg/kg and 40 mg/kg group,which was significantly different from that of AR model group (2 weeks TSLP: (0.897 8±0.081 8)/(1.072 1±0.113 6)/(1.396 6±0.133 9) vs 1.795 9±0.131 4; 4 weeks TSLP: (1.191 0±0.161 3)/(1.141 0±0.152 3)/(1.200 5±0.189 6) vs 1.809 7±0.253 4; 2 weeks TSLP-mRNA: (1.175 6±0.100 9)/(1.254 4±0.078 2)/(2.037 2±0.559 2) vs 4.582 9±0.697 7; 4 weeks TSLP-mRNA: (1.158 3±0.104 3)/(1.224 0±0.034 0)/(1.275 2±0.099 6) vs 4.814 4±0.662 8; all P<0.05), and not significantly different from control group. With the inhibition of TSLP, the concentrations of IL-4 and OVA-sIgE in rat serum were also decreased. Conclusion 18β-sodium glycyrrhetinic acid has obvious inhibitory effect on TSLP in nasal mucosa of AR rats, which can control Th2 type immune inflammatory reaction.