ABSTRACT
In this study, we investigated the effect of latanoprost on the expression of TGF- ß1 and Wnt / ß - Catenin signal pathway in the choroid of form-deprivation myopia model rats. Forty rats were randomly divided into two groups: the control group and the FDM model group. Each group had 20 rats. The FDM model group was established by feeding latanoprost daily for 28 days. 15 rats in each group were used to measure the length of the ocular axis and the level of TGF-ß1 in choroidal tissue; the remaining 5 rats in each group were used for choroidal fibroblast culture. After modeling, the rats were killed, the length of the ocular axis was measured with a vernier caliper, and the level of TGF - ß1 protein and mRNA in the choroidal tissue of each group were measured with RT-PCR method. Results showed that compared with the control group, there was a significant difference in the axial length of the FDM model group (P< 0.05). There was a significant difference in the expression of TGF- ß1 protein and mRNA between the two groups (P<0.05). The cultured cells were identified as choroidal fibroblasts by immunocytochemistry. There was no significant difference (P>0.05) in the comparison of GSK3 ß protein in choroidal fibroblasts of rats in each group. TGF-ß 1 and APC protein in FDM group were significantly lower than those in the control group (P<0.05), while dcl3, p21-gsk3 ß and ß - Catenin proteins were significantly higher (P<0.05), and there was no significant difference (P>0.05) in the ratio of various indexes protein in FDM + ddk1 group and the comparison of TGF - ß1 and APC protein in FDM + ddk1 group and FDM group The expression of dcl3, p21-gsk3 ß and ß - Catenin decreased significantly (P<0.05). There was no significant difference in the expression of GSK3 ß mRNA in the choroidal fibroblasts of each group (P>0.05). The expression of TGF - ß 1 and APC mRNA in FDM group was significantly lower than that in the control group (P<0.05), while the expression of dcl3, p21-gsk3 ß and ß-catenin mRNA in FDM + ddk1 group was significantly higher than that in the control group (P<0.05) >In FDM + ddk1 group, TGF-ß 1 and APC mRNA were significantly lower than those in FDM group (P<0.05), while dcl3, p21-gsk3 ß and ß-Catenin mRNA were significantly higher (P<0.05).
Subject(s)
Choroid/drug effects , Latanoprost/pharmacology , Myopia/drug therapy , Transforming Growth Factor beta1/metabolism , Wnt Signaling Pathway/drug effects , beta Catenin/metabolism , Animals , Cells, Cultured , Choroid/metabolism , Female , Fibroblasts/drug effects , Fibroblasts/metabolism , Male , Myopia/metabolism , RNA, Messenger/metabolism , Rats , Rats, Sprague-DawleyABSTRACT
OBJECTIVE:To establish a method for the simultaneous determination of loganic acid and isoscoparin in Sanwei longdanhua tablets. METHODS:HPLC method was adopted. The determination was performed on Inertsil ODS-3 column with mobile phase consisted of methanol-0.2% phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 240 nm,and column temperature was 30 ℃. The sample size was 10 μL. RESULTS:The linear range were 0.040 08-4.008 0 μg(r＝0.999 9)for loganic acid and 0.021 96-2.196 0 μg(r＝0.999 9)for isoscoparin. The quantitative limits were 0.160 32 and 0.087 8 ng/mL,and detection limits were 0.080 16 and 0.043 92 ng/mL. RSDs of precision,stability and reproducibility tests were all lower than 2%. The recoveries were 103.07%-104.26%(RSD＝0.52%,n＝6) and 95.57%-99.61%(RSD＝1.55%,n＝6). CONCLUSIONS:The method is simple,accurate and suitable for simultaneous determination of loganic acid and isoscoparin in Sanwei longdanhua tablets.