ABSTRACT
Objective To observe the effects of Qijingmingmu decoction granule on the expression of mitogen-activated protein linase (MAPK) signal pathway in fibroblasts of conjunctivochalasis (CCH) under the stimulation of tumor necrosis factor (TNF-α) and to explore the pathogenesis and effective treatment method of CCH.Methods CCH conjunctival fibroblasts were cultured in vitro and divided into CCH group,CCH + TNF-α group and CCH + TNF-α + Qijingmingmu decoction group.CCK-8 assay was used to determine the effective concentration of Qijingmingmu decoction granule,and 10-2 mg · L-1 TNF-α was added in the cultured fibroblasts of the latter two groups,followed by interference with Qijingmingmu decoction granule for 48h.The expression of MAPK signal pathway related protein and mRNA were detected by ELISA,Western Blot and RT-PCR.Then the results were statistically analyzed.Results CCK-8 assay showed the effective concentration of Qijingmingmu decoction granule was 1.61g · L-1.And there were significant differences in the total A values (A450) of extracellular regulated protein kinase (ERK),c-jun N-terminal kinase (JNK),p38 mitogen activated protein kinase (p38 MAPK) and their phosphorylation levels among the three groups (all P ＜ 0.05).Moreover,TNF-α could significantly up-regulate the expression of ERK1/2,JNK1/2,p-JNK1/2,p38 MAPK and p-p38 MAPK in CCH fibroblasts (all P ＜0.05),while Qijingmingmu decoction down-regulated their expressions in CCK fibroblasts after TNF-α stimulation (all P ＜ 0.05).Furthermore,the total differences in p-ERK1/2,p38 MAPK and p-p38 MAPK protein were significant in the three groups (all P＜0.05);and TNF-α could significantly up-regulate the expression of p-ERK1/2,p-JNK1/2,p38 MAPK and p-p38 MAPK protein in CCH fibroblasts (all P ＜0.05),while Qijingmingmu decoction down-regulated p-ERK1/2 and p38 MAPK expressions in CCK fibroblasts after TNF-α stimulation (both P ＜ 0.05).In addition,the total differences in ERK1/2 and p38 MAPK mRNA were significant in the three groups (both P ＜ 0.05);and TNF-α could significantly up-regulate the expression of ERK1/2 and p38 MAPK mRNA in CCH fibroblasts (both P ＜ 0.05),while Qijingmingmu decoction down-regulated p38 MAPK mRNA expressions in CCK fibroblasts after TNF-α stimulation (P ＜ 0.05).Conclusion The inflammatory factor TNF-α can up-regulate the expressions of MAPK signal pathway related protein and mRNA in CCH fibroblasts,resulting in the occurrence and development of CCH,and meanwhile Qijingmingmu decoction granule can downregulate the expression of MAPK signal pathway to play the therapeutic role in CCH to some extent.
ABSTRACT
·AIM:To compare the similarities and differences of central corneal thickness (CCT) measured by different devices, so as to provide a safe, effective, simple and accurate method for measuring corneal thickness. ·METHODS: Totally 95 eyes of 95 cataract patients enrolled continuously for preoperative examination in ophthalmology department of Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from September 2016 to December 2016. Handheld automatic PachPen ultrasonic pachymetry (USP), anterior segment optical coherence tomography (AS-OCT) and specular microscopy were respectively used to measure the central corneal thickness,and intraocular pressure was measured at the same time. The similarities and differences of these three different devices to measure the central corneal thickness and their correlations with intraocular pressure were observed. Then the data were analyzed statistically. · RESULTS: The CCT value measured by PachPen ultrasonic pachymetry was 544. 43 ± 36. 61μ m, the CCT value measured by AS-OCT was 527.09 ± 35.54μ m, and the CCT value measured by specular microscopy was 533.20 ± 30. 17μ m. There was significant difference between these three groups(F=6.272,P=0.002),and the CCT value measured by PachPen ultrasonic pachymetry was significantly higher than the other two groups. The correlation coefficients between the CCT values of these three groups and intraocular pressure were 0.290, 0.277 and 0.204 (P<0.05) respectively, of which the correlation between the CCT measured by PachPen ultrasonic pachymetry and the intraocular pressure was the highest. There was statistically significant correlation between the CCT values measured by these three measurements (P<0.001). ·CONCLUSION: The CCT value measured by PachPen ultrasonic pachymetry is the highest. The second CCT value is measured by specular microscopy, and the smallest CCT value is measured by AS-OCT. Obvious correlation and good consistency were found in the CCT values measured by three types of devices, and the CCT values were all correlated with intraocular pressure. Therefore,we should take more attention in clinic.