ABSTRACT
Objective To construct a recombinant adenovirus vector expressing mouse SPINK5 gene,and observe its curative effect on the skin lesions in atopic dermatitis mice model. Methods By recombining DNA technology,the sequence of mouse SPINK5 gene was cloned into adeno?virus shuttle plasmid. Then it was transformed into HEK 293 cells with the adenoviral backbone plasmid to obtain the recombinant adenovirus. A mouse model of atopic dermatitis was established by system and local sensitization of Balb/c mice with ovalbumin . The effect of recombinant adeno?virus on the lesions of atopic dermatitis mice model was observed. Results The SPINK5 over?expressing adenovirus vector and atopic dermatitis mice model were successfully constructed. After 2 weeks of adenovirus?mediated SPINK5 gene intracutaneous injection,the redness and edema of lesions of AD model mice were obvious relieved. The pathological detection indicated that epidermal thickness and prickle cell layer ,inflammatory cell infiltration significant decreased accompanied with the model blank control. Conclusion The adenovirus?mediated SPINK5 gene had signifi?cant therapeutic effect to the atopic dermatitis mice model ,which provided a laboratory basis of application of SPINK5 gene product to therapy atopic dermatitis.
ABSTRACT
Objective To investigate the influence of the eye acupuncture therapy in the expressions of brain derived neurophic factor(BDNF)and tyrosine kinase receptor B(TrkB)in penumbra brain tissue of ischemic area in the rats with acute cerebral ischemia reperfusion inj ury,and to explore the related mechanism of protective effect on brain of eye acupunture.Methods 62 SD rats were randomly divided into blank control group (n=11),sham operation group (n=11)and model copy group (n=40).The rats in copy model group were used to establish models with suture method,and the successful model rats (n=32)were randomly divided into model group(n=16)and eye acupuncture group(n=16).The rats in eye acupuncture group were treated with eye acupuncture intervention;the acupoints were chosen according to human acupoint selection method, the liver, the upper-jiao, the down-jiao district,and renal were selected for acupuncture intervention. After reperfusion for 2 h,the acupuncture was performed once every 8 h,lasted for 10 times;30 min after the last intervention,the rats were sacrificed;the regional ischemia penumbra around the parts of the brain tissue was obtained and the expressions of BDNF and TrkB mRNA and protein in brain tissue of the rats were detected with RT-PCR and Western blotting method. Results Compared with blank control group,the expression levels of BDNF and TrkB mRNA and protein in brain tissue of the rats in sham operation group had no statistical significance(P>0.05);compared with sham operation group,the expression levels of BDNF and TrkB mRNA and protein in brain tissue of the rats in model group were significantly up-regulated(P<0.01);compared with model group,the expression levels of BDNF and TrkB mRNA and protein in brain tissue of the rats in eye acupuncture group were significantly up-regulated(P<0.05 or P<0.01).Conclusion The expressions of BDNF and TrkB are increased after cerebral ischemia and reperfusion injury.Eye acupuncture can protect the brain of MCAO/R rats,which may be related to the up-regulation of the expressions of BDNF and TrkB in the corresponding penumbra.
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Objective To observe the effects of Grifola frondosa polysaccharide on T cell subsets and Th1/Th2 subsets of spleen deficiency mice, and investigate its immunoregulation mechanism. Methods Forty-eight KM mice were divided into 6 groups:normal group, spleen deficiency group, positive group (lentinan), three dosage groups of Grifola frondosa polysaccharide, 8 mice in each group. The mice were injected corresponding drug intraperitoneally for 10 days. Then mice in each group were detected CD3+, CD4+ and CD8+T cell by flow cytometry. IFN-γ and IL-4 levels were detected by ELISA. Results The high dose of Grifola frondosa polysaccharide could significantly increase the CD4+T lymphocytes percentage in peripheral blood (P<0.05). Medium and high dose of Grifola frondosa polysaccharide could significantly increase CD4+/CD8+T cell ratio (P<0.01) and CD3+T lymphocyte percentage (P <0.01) of spleen deficiency mice. Medium and high dose of grifola frondosa polysaccharide could significantly increase IFN-γ level in serum of spleen deficiency mice (P<0.01), and high dose of Grifola frondosa polysaccharide could decrease IL-4 level in serum of spleen deficiency mice (P<0.01). Conclusion Grifola frondosa polysaccharide can improve the decreased CD4+/CD8+T cell ratio which caused by spleen deficiency, and promote the transformation of Th1 to Th2, thus treating spleen deficiency syndrome.
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Objective To explore the features of immune responsed activated by polysaccharides from herbal medicines and provide basis for immunomodulation of polysaccharides. Methods The mice were immuned by polysaccharides of Angelica, Atractylodes and Giant typhonium rhizome respectively, then the corresponeding antibodies and cross-reactive antibodies in serum were detected by ELISA. Results Three kinds of polysaccharides from herbal medicines could all activated the mice to produce not only the corresponding antibodies, but also the cross-reactive antibodies, the ranges of non-specific antibodies activated by distinct polysaccharide were different. Conclusions Polysaccharides from herbal medicines may be a kind of non-specific and broad-spectrum immunomodulators.