1.Effects of 13-hexyl-berberine hydroehloride and 13-hexyl-palmatine hydroehloride on the activation of nuclear factor-kappa B and phosphorylation of p38 mitogen-activated protein kinase (MAPK) in a human keratinooyte cell line, Hat?aT stimulated by tumor n
Hui JIANG ; Xinyu LI ; Hongqin TU ; Yongfang WANG ; Shasha SONG ; Lanfang XU
Chinese Journal of Dermatology 2009;42(5):339-342
Objective To investigate the effects of 13-hexyl-berbefine hydroehlofide (HB-13) and 13-hexyl-paimatine hydrochloride (HP-13) on the activation of nuclear factor-kappa B (NF-kB) and phosphorylation of p38 mitogen-activated protein kinase (MAPK) in a human keratinocyte cell line, HaCaT stimulated by tumor necrosis factor alpha (TNF-α). Methods HaCaT cells were cultured in the presence of various concentrations (0.39, 0.78, 1.56 μg/mL) of HB-13 or HP-13 for 120 minutes followed by the stimulation with recombinant human TNF-α for 120 minutes (in phosphorylatEd-IkB-α test) or 15 minutes (in phosphorylated-p38 test). Then, HaCaT cells were disrupted, total protein was extracted, and the expressions of phosphorylated I B-α and phosphorylated p38 were detected with Western blot. HaCaT cells receiving neither pretreatment nor stimulation served as blank control, untreated HaCaT cells stimulated by rhTNF-α as stimulator control, and HaCaT cells pretreated with turmeric root tuber and stimulated by rhTNF-α as positive control. Results From 0.39 to 1.56 μg/mL, both HB-13 and HP-13 significantly inhibited the expression of p-IkB-α in HaCaT cells stimulated by rhTNF-α, and a nonsignificant dose-dependent trend was observed for their inhibitory effect, with the ICo value being 0.441 μg/mL for I-IB-13 (r = -0.990, n = 3, P > 0.05) and 0.832 μg/mL for HP-13 (r = -0.992, n = 3, P > 0.05). In contrast, neither 1-113-13 nor HP-13 within the experiment concentration range had a significant effect on the expression of p-p38 in HaCaT cells stimulated by rhTNF-α (P > 0.05). Conclusions Within the experimental concentration range, both HB-13 and HP-13 can inhibit the activation of NF-kB in HaCaT cells induced by TNF-α signal, but neither of them suppress the phosphorylation of p38MAPK induced by TNF-α signal in HaCaT cells.
2.Effects of triptolide on interferon-γ signaling in a human keratinocyte cell line HaCaT
Hongqin TU ; Xinyu LI ; Heng GU ; Hui JIANG ; Lanfang XU ; Yongfang WANG ; Shasha SONG
Chinese Journal of Dermatology 2009;42(3):167-170
Objective To investigate the effects of triptolide on the expression of a series of proteins associated with interferon-γ (IFN-γ)signaling in HaCaT keratinocytes.Methods After pretreatment with difrerent dosages of triptolide(10-10-10-7 mol/L),HaCaT cells were stimulated by recombinant human IFN-γ(rhIFN-γ,500 U/mL)for various periods followed by the collection of cells.Then,total protein was extracted from these cells and subjected to Western blotting for the detection of expression of interferon-γ receptor α(IFN-γRα),phosphorylated Janus kinase 2(pJAK2)and suppressor of cytokine signaling (SOCS1).Results Triptolide at the concentrations of 10-8 mol/L and 10-7 mol/L significantly inhibited the IFN-γRα expression upregulated by rhIFN-γ(both P<0.05).The expression of pJAK2 induced bv rhIFN-γ was also suppressed by triptolide at the concentrations of 10-9 moI/L and 10-8 mol/L(both P<0.05).The inhibition of triptolide on IFN-γRα and pJAK2 expression was dose-dependent and the 50%inhibitory concentrations(IC50 value)were 1.37×10-8 mol/L and 2.83×10-9 mol/L,respectively.On the contrary,triptolide upregulated the expression of SOCS1 stimulated by rhIFN-γ at the concentrations of 10-10,10-9 and 10-8 mol/L(P<0.05,0.05,0.01,respectively)with the 50%effective dosage(ED50 value)at 3.32 × 10-11 mol/L.Conclusions By inhibiting the expression of IFN-γRα as well as phosphorylation of JAK2 and upregulating the expression of SOCS1,triptolide inhibits the phosphorylation of STAT-1,resulting in the inhibition of genetic transcription of multiple inflammatory factors induced by IFN-γ signaling in HaCaT keratinocytes,and the inhibition probably contributes to the efficacy of triptolide in the treatment of IFN-γ-dependent inflammatory skin disorders,such as psoriasis.
3.Changes in refractive power and ocular biometrics before and after the onset of myopia in children: the Anyang Childhood Eye Study
Menghai SHI ; Ying HUANG ; He LI ; Yongfang TU ; Lei LI ; Mengtian KANG ; Shifei WEI ; Yunyun SUN ; Lei YIN ; Ningli WANG ; Shiming LI
Chinese Journal of Experimental Ophthalmology 2024;42(5):453-461
Objective:To analyze the trends in refractive error and ocular biological parameters in elementary school students over 5 years, and to investigate the patterns of change before and after myopia onset.Methods:A cohort study was adopted.A total of 1 986 first-grade students from the Anyang Childhood Eye Study were enrolled in this cohort study and their right eye data were taken for analysis, including 1 126 boys and 860 girls.Every year, cycloplegic autorefraction was performed with 1% cyclopentolate eyedrops to obtain the spherical equivalent (SE).The axial length (AL), anterior chamber depth, lens thickness, mean corneal curvature (Km) and other parameters were obtained by ocular biometry.The lens refractive power (LP) was calculated using the Bennett formula.The subjects were assigned to persistent myopia group, non-myopia group and new onset myopia group.According to the age of myopia onset, the new onset myopia group was subdivided into the 8-, 9-, 10-, 11- and 12-year-old myopia groups to compare the differences in refractive error and ocular bioparameters among groups at different time points of follow-up.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Beijing Tongren Hospital, Capital Medical University (No.TRECKY2018-030).Written informed consent form was obtained from the guardians of each subject.Results:All children had a gradual SE drift toward myopia and a gradual increase in the AL with age, and there were significant differences in SE and AL between adjacent follow-up ages within the three groups (all at P<0.05).The earlier the onset of myopia, the higher the myopia SE and the longer the AL of the eye at the same follow-up age, the differences in SE between adjacent groups were statistically significant (all at P<0.05), and the differences in AL between adjacent groups at the follow-up age of 8 to 12 years were statistically significant (all at P<0.05).In the nonmyopia group, SE drifted toward emmetropia at a slow and steady rate of (-0.23±0.27)D/year, and AL also increased slowly and steadily at (0.18±0.13)mm/year.In the new onset myopia group, the changes in SE in the third, second, and first years before myopia onset were (-0.32±0.25), (-0.45±0.33), and (-0.98±0.44)D, and the increases in AL were (0.25±0.12), (0.32±0.15), and (0.48±0.19)mm, respectively.Both SE and AL change rates began to accelerate before myopia onset and slowed down after myopia onset, with statistically significant differences in the overall comparison of SE and AL change rates at different time intervals before and after myopia onset (all at P<0.001).The AL at myopia onset in boys was (24.11±0.70)mm, which was longer than (23.60±0.66)mm in girls ( t=159.71, P<0.01).LP decreased with age in all groups, with a faster rate before the age of 9 years and a slower rate after the age of 9 years.The mean decrease rate in LP was (-0.48±0.19), (-0.44±0.20), (-0.49±0.16), (-0.51±0.18), and (-0.48±0.19)D/year in the persistent myopia group and 8~11-year-old myopia group, respectively, which were significantly faster than -0.42±0.17 D/year in 12-year-old myopia group and (0.37±0.15)D/year in nonmyopia group (all at P<0.05).There was no statistically significant difference in Km among groups at different follow-up ages (all at P>0.05). Conclusions:The AL begins to grow at an accelerated rate 3 years before myopia onset, and the increase rate of the AL slows down after the onset of myopia, but it is still significantly faster than that of non-myopic children.In this process, the decrease in LP plays a compensatory role; there is no significant change in corneal curvature.The AL of males at the onset of myopia is longer than that of females at the same age.AL is an important indicator for the prevention and control of myopia.It is important to consider gender differences and to pay more attention to the growth rate when assessing AL.