ABSTRACT
Objective:To investigate the effect of Xidi Liangxue recipe on the proliferation and apoptosis of HaCaT cells through the long non-coding RNA (lncRNA) nuclear-enriched abundant transcript 1 (NEAT1) /microRNA (miR) -485-5p/signal transducer and activator of transcription 3 (STAT3) regulatory network. Methods:HaCaT cells were induced by interleukin-17 (IL-17), and the mRNA and protein expression of lncRNA NEAT1, miR-485-5p and STAT3 was detected in IL-17-induced HaCaT cells and normal human epidermal keratinocytes (NHEK) by quantitative PCR (qPCR) and Western blot analysis, respectively. The location of lncRNA NEAT1 and miR-485-5p in IL-17-induced HaCaT cells was observed by fluorescence in situ hybridization (FISH), and the targeted regulatory relationship among lncRNA NEAT1, miR-485-5p and STAT3 was verified by double-luciferase reporter gene assay. Chinese herbs were decocted according to the Xidi Liangxue recipe, SD rats were divided into two groups to be gavaged with the above decoctions (medicated group) or physiological saline (control group) for 5 days, and then serum samples were collected from the above two groups of rats separately. The IL-17-induced HaCaT cells were divided into 4 groups: control group treated with the control sera, lncRNA-NEAT1 overexpression group transfected with lncRNA-NEAT1 overexpression vectors and treated with the control sera, Xidi Liangxue recipe group treated with the medicated sera, and Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group transfected with lncRNA-NEAT1 overexpression vectors and treated with the medicated sera. qPCR, Western blot analysis, flow cytometry, and cell counting kit (CCK8) assay were performed to determine the mRNA and protein expression of lncRNA NEAT1, miR-485-5p and STAT3, and to evaluate cell proliferation and apoptosis. The two independent samples t-test was used for comparisons between two groups, one-way analysis of variance for comparisons among multiple groups, and least significant difference (LSD) t-test for multiple comparisons. Results:The IL-17-induced HaCaT cell group showed significantly increased relative expression levels of lncRNA NEAT1 and STAT3 mRNA (1.84 ± 0.21, 2.20 ± 0.24, respectively) and significantly increased protein expression of STAT3 and p-STAT3 (1.27 ± 0.13, 2.43 ± 0.16, respectively), but significantly decreased expression level of miR-485-5p (0.32 ± 0.04) compared with the NHEK group (lncRNA NEAT1 and STAT3 mRNA: 1.00 ± 0.11, 1.00 ± 0.11, respectively, both P < 0.05; STAT3 and p-STAT3 protein: 1.00 ± 0.11, 1.00 ± 0.10, t = 2.54, 3.02, respectively, both P < 0.05; miR-485-5p: 1.00 ± 0.12, t = 2.94, P = 0.015). FISH demonstrated that miR-485-5p and lncRNA NEAT1 were co-located in the cytoplasm of HaCaT cells. The double-luciferase reporter gene assay showed that the relative activity of luciferase was significantly lower in the miR-485-5p group than in the negative control group (both P < 0.05) after the transfection with wild-type lncRNA NEAT1 or STAT3 recombinant plasmids, while there were no significant differences between the miR-485-5p group and negative control group after the transfection with mutant lncRNA NEAT1 or STAT3 recombinant plasmids (both P > 0.05). Compared with the control group, the lncRNA-NEAT1 overexpression group showed significantly increased expression of lncRNA NEAT1 and STAT3 (including STAT3 mRNA, STAT3 protein, and p-STAT3 protein) in HaCaT cells (all P < 0.05), but significantly decreased miR-485-5p expression ( P < 0.05) ; the Xidi Liangxue recipe group showed significantly decreased expression of lncRNA NEAT1 and STAT3 (all P < 0.05), but significantly increased miR-485-5p expression compared with the control group ( P < 0.05) ; significantly decreased expression of lncRNA NEAT1 and STAT3, but significantly increased miR-485-5p expression was observed in the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group compared with the lncRNA-NEAT1 overexpression group (all P < 0.05). After 24-, 48-, and 72-hour intervention, CCK8 assay showed that the proliferative activity of HaCaT cells was significantly higher in the lncRNA-NEAT1 overexpression group than in the control group (all P < 0.05), as well as in the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group than in the Xidi Liangxue recipe group (all P < 0.05), and the cellular proliferative activity was significantly lower in the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group and Xidi Liangxue recipe group than in the control group (all P < 0.05). The apoptosis rate was significantly lower in the lncRNA-NEAT1 overexpression group (5.84% ± 0.28%) than in the control group (14.75% ± 0.83%, LSD- t = 3.48, P = 0.002), but significantly higher in the Xidi Liangxue recipe group (35.72% ± 3.62%) than in the control group (LSD- t = 5.34, P = 0.001) ; the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group showed significantly increased apoptosis rate (27.64% ± 2.82%) compared with the lncRNA-NEAT1 overexpression group (LSD- t = 9.06, P < 0.001) . Conclusion:The Xidi Liangxue recipe could inhibit the proliferation of IL-17-induced HaCaT cells and promote their apoptosis, which may be related to the intervention in the lncRNA NEAT1/miR-485-5p/STAT3 regulatory network.
ABSTRACT
Objective To investigate the clinical efficacy and safety of 308-nm excimer laser combined with tazarotene gel for the treatment of plaque psoriasis.Methods Seventy-two patients with plaque psoriasis were randomly and equally divided into three groups according to a random number table:tazarotene group topically applying tazarotene gel once per night,308-nm excimer laser group treated with 308-nm excimer laser,combination group treated with both tazarotene gel and 308-nm excimer laser.Clinical efficacy was evaluated according to psoriasis area and severity index (PASI) score and response rate,and safety according to adverse reactions at week 2,4 and 8 after starting treatment.Results PASI score was significantly lower in the combination group at week 4 and 8 (4.75 ± 0.44 and 2.35 ± 0.37 respectively) than in the 308-nm excimer laser group (6.75 ± 0.57 and 4.67 ± 0.36 respectively,both P ＜ 0.05) and tazarotene group (8.75 ± 0.48 and 6.48 ± 0.45 respectively,both P ＜ 0.05),and significantly lower in the combination group at week 8 than at week 2 and 4 (both P ＜ 0.05).A significant increase was observed in the response rate at week 2,4,and 8 in the combination group (29.1％ (7/24),66.7％ (16/24) and 87.5％ (21/24) respectively) compared with the tazarotene group (12.5％ (3/24),41.7％ (10/24) and 62.5％ (15/24) respectively,all P＜ 0.05) and 308-nm excimer laser group (20.8％ (5/24),50.0％ (12/24) and 75.0％ (18/24) respectively,all P＜ 0.05).No systemic adverse reactions were observed in any of the 3 groups during the study,and there was no significant difference in the incidence of local adverse reactions between the combination group,tazarotene group and 308-nm excimer laser group (16.7％ (4/24) vs.12.5％ (3/24) vs.12.5％ (3/24),P ＞ 0.05).Conclusion The efficacy of 308-nm excimer laser combined with tazarotene gel is superior to that of tazarotene gel or 308-nm excimer laser alone in the treatment of plaque psoriasis.
ABSTRACT
Objective To evaluate the efficacy of Xuebijing injection plus oral acitretin for the treatment of erythrodermic psoriasis.Methods Forty-eight patients with erythrodermic psoriasis were equally and randomly divided into two groups by a random number table:test group treated with Xuebijing injection once a day plus oral acitretin,and control group treated with oral acitretin.The dose of acitretin began at 0.5 mg per kilogram per day,and was modified according to the tolerance in and response of patients.After 8 weeks of treatment,clinical efficacy was evaluated by psoriasis area and severity index (PASI) score,response rate and recurrence rate.Adverse reactions were also recorded and evaluated.Results The difference in PASI score between pre-and post-treatment was significantly higher in the test group than in the control group (34.9 ± 2.2 vs.27.3 ± 1.7,t =3.37,P ＜ 0.05).The total response rate was 87.5％ in the test group and 62.5％ in the control group (x2 =4.87,P ＜ 0.05).There was a statistical decrease in the average onset time ((13.5 ± 2.4) d vs.(20.7 ± 3.1) d,t =3.67,P ＜ 0.05),daily dose and total dose of acitretin ((26.4 ± 3.3) mg vs.(34.7 ± 3.5) mg,(1854.5 ± 85.2) mg vs.(2768.8 ± 88.7) mg,t =3.07,4.32,respectively,both P ＜ 0.05) in the test group compared with the control group.The recurrence rate was 9.5％ (2/21) in the test group and 26.6％ (4/15) in the control group (x2 =5.23,P ＜ 0.05).Conclusion In the case of erythroderma psoriaticum,Xuebijing injection combined with oral acitretin is superior to oral acitretin alone in clinical efficacy,onset time and reducing recurrence.