ABSTRACT
Objective To investigate hydrogen peroxide (H2O2) and transforming growth factor-β2 (TGF-β2) induced fibronectin (FN), collagen 1 (COL1), nuclear factor (NF)-κB P65 proteins and interlukin (IL)-1βgene expression in human trabecular meshwork cells (HTMCs), and the interventional mechanism of resveratrol (RSV). Methods (1) HTMCs with 70 to 80%confluency were divided into 5 groups. The experimental groups were treated with serum-free medium and with H2O2 at concentrations of 150, 300, 450 and 800μmol/L. The control group was treated with 0μmol/L H2O2. The protein levels of FN, COL1, NF-κB P65 and NF-κB P65 phosphorylation (P-NF-κB P65) were measured by Western blot assay. The expression of IL-1βgene was measured by qPCR. (2) HTMCs were divided into 3 groups. The control group was treated withserum-free medium and without H2O2 and RSV. The H2O2 group was treated with 300μmol/L H2O2. The H2O2+RSV group was treated with 300μmol/L H2O2 and 25μmol/L resveratrol (RSV). The expressions of proteins and genes mentioned above were detected in three groups. NF-κB P65 nuclear translocation was assessed by immunofluorescence technique. (3) HTMCs were divided into 3 groups. The control group was treated with serum-free medium and without TGF-β2 and RSV. The TGF-β2 group was treated with 5μg/L TGF-β2. The TGF-β2+RSV group was treated with 5μg/L TGF-β2 and 25μmol/L RSV. The expressions of proteins and genes mentioned above were detected in three groups. Results (1) Compared with control group, the protein levels of FN and P-NF-κB P65 were significantly increased in 150, 300, 450 and 800μmol/L groups,the expression levels of COL1 protein and IL-1β gene were significantly increased in 300, 450 and 800 μmol/L groups (P <0.05). There were no statistical significances between other indicators. (2) The expression levels of FN, COL1, P-NF-κB P65 proteins and IL-1βgene were significantly higher in H2O2 group than those in control group, and which were significantly lower in H2O2+RSV group than those in H2O2 group. Compared with control group, only the expression of IL-1βgene was decreased in H2O2+RSV group (P < 0.05). NF-κB P65 was only expressed in cytoplasm in control group, while it was expressed in both cytoplasm and nucleus in H2O2 group. Compared with H2O2 group, NF-κB P65 was mainly expressed in cytoplasm. (3) Compared with control group, the expressions of FN, COL1, P-NF-κB P65 proteins and IL-1β gene were significantly increased in TGF-β2 group (P < 0.05). Compared with TGF-β2 group, the indicators mentioned above were significantly decreased in TGF-β2+RSV group (P<0.05). Conclusion H2O2 and TGF-β2 can upregulate the expression of FN, COL1, P-NF-κB P65 proteins and IL-1βgene in HTMCs, which may be involved in the development and progression of glaucoma. RSV can inhibit the influence of H2O2 and TGF-β2 in HTMCs and exert a protective effect on glaucoma.
ABSTRACT
The efferent and afferent projections of the hyperstriatum ventrale, pars caudal nucleus (HVc) in carduelis spinus were investigated using anterograde and retrograde HRP tracing techniques. After injecting HRP into the HVc, anterogradely labelled fibers and terminals were observed in the robust nucleus of the archistriatum (RA), tractus archistriatum dorsalis (DA) and the area X of lobus parolfactorius. The dense retrogradely labelled cell bodies were found in medial magnocellular nucleus of the anterior neostriatum (mMAN), nucleus interfacialis (NIf) of midneostriatum, telencephalic auditory area--field L of the neostriatum, and locus ceruleus (LoC) of pons. All these projections were ipsilateral. Based on the present and previous studies, the authors got the following understanding: 1. The HVc projects to the high vocal control centre--RA. This proved that the HVc is also a high vocal control nucleus. 2. The HVc receives afferent projections from the telencephalic auditory area--field L. It indicates that there are direct connection between the two high centres of audition and vocal control nuclei in telencephalon. 3. This study demonstrated that the HVc receives afferent projection from the LoC for the first time. According to the fact, we presume that HVc may be involved in the neural regulations of vegetative function and emotional reactions. 4. The HVc also receives afferent projections from the mMAN and NIf of neostriatum, and from the Uva of the thalamus. On the other hand the HVc projects to area X. Since the mMAN, NIf, Uva and area X are vocal learning and memory centres, therefore the HVc may be also involve in vocal learning and memory functions.