1.Circ_0086376 targeted miR-5195-3p to regulate the expression of inflammatory factors mediated by Propionibacterium acnes in HaCaT cells
Ruixian YE ; Rujun XUE ; Jingyao LIANG ; Xibao ZHANG
Chinese Journal of Dermatology 2025;58(4):340-346
Objective:To investigate the effects of circular RNA circ_0086376 on the expression of acne-related inflammatory factors by targeting microRNA (miR) -5195-3p.Methods:Circ_0086376-overexpressing or -interferred stable HaCaT cell lines were constructed and co-cultured with Propionibacterium acnes ( P.acne). Quantitative real-time PCR was used to detect the effect of co-culture on circ_0086376 expression, while enzyme-linked immunosorbent assay (ELISA) was performed to measure the expression of inflammatory factors in the cell culture supernatant. The overexpression or interference of circ_0086376 plasmid and lentivirus mimicking or inhibiting miR-5195-3p fragment were transfected into HaCaT cells, and ELISA was used to detect the expression of inflammatory factors in the culture supernatant of HaCaT cells after overexpression or interference of circ_0086376 cultured alone or with P.acne. The luciferase reporter assay was conducted to verify the targeting relationship between circ_0086376 and miR-5195-3p. Additionally, ELISA was used to detect the effects of overexpression/interference of circ_0086376 and mimic/inhibition of miR-5195-3p on the expression of inflammatory factors in the culture supernatant of HaCaT cells co-cultured with P.acne. Results:After co-culture with P.acne, the levels of inflammatory factors in the culture supernatant were significantly higher than those in the HaCaT cells cultured alone, including Interleukins (IL) -1β ([355.80 ± 23.20] vs. [260.50 ± 16.58] pg/ml, t = 5.79, P < 0.01), IL-6 ([38.04 ± 2.69] vs. [14.33 ± 0.75] pg/ml, t = 14.65, P < 0.01), IL-12 ([10.87 ± 0.78] vs. [6.52 ± 0.77] pg/ml, t = 6.89, P < 0.01), IL-18 ([222.60 ± 21.07] vs. [146.10 ± 9.14] pg/ml, t = 5.77, P < 0.01), and Tumor necrosis factor (TNF) -α ([50.39 ± 1.29] vs. [20.46 ± 0.83] pg/ml, t = 33.83, P < 0.01). The expression levels of IL-1β, IL-6, IL-12, IL-18 and TNF-α in the culture supernatant of HaCaT cells in the circ-empty vector + P.acne group were higher than those in the circ-empty vector overexpression group. The levels of inflammatory factors mentioned above in circ-overexpression + P.acne group were lower than those in circ-overexpression empty vector + P.acne group ( P < 0.01). The expression levels of inflammatory factors mentioned above in the culture supernatant of HaCaT cells in the interference circ-empty vector + P.acne group were higher than those in the interference circ-empty vector group. Compared with the interference circ-empty vector + P.acne group, the expression of circ in the interference circ + P.acne group was higher ( P < 0.01). Luciferase reporter assay confirmed that circ_0086376 could bind to miR-5195-3p. The expression levels of IL-1β, IL-6, IL-12, IL-18 and TNF-α in the circ-overexpression group were lower than those in the empty vector group ( P < 0.05), and the levels of these inflammatory factors in the circ-overexpression + miR mimic group were higher than those in the circ overexpression group ( P < 0.05). The expression levels of inflammatory factors in the interference circ group were higher than those in the empty vector group, levels of inflammatory factors in the circ + miR-inhibiting group were lower than those in the circ interference group ( P < 0.05) . Conclusion:Circ_0086376 can inhibit the expression of acne-related inflammatory factors by targeting and downregulating miR-5195-3p.
2.Circ_0086376 targeted miR-5195-3p to regulate the expression of inflammatory factors mediated by Propionibacterium acnes in HaCaT cells
Ruixian YE ; Rujun XUE ; Jingyao LIANG ; Xibao ZHANG
Chinese Journal of Dermatology 2025;58(4):340-346
Objective:To investigate the effects of circular RNA circ_0086376 on the expression of acne-related inflammatory factors by targeting microRNA (miR) -5195-3p.Methods:Circ_0086376-overexpressing or -interferred stable HaCaT cell lines were constructed and co-cultured with Propionibacterium acnes ( P.acne). Quantitative real-time PCR was used to detect the effect of co-culture on circ_0086376 expression, while enzyme-linked immunosorbent assay (ELISA) was performed to measure the expression of inflammatory factors in the cell culture supernatant. The overexpression or interference of circ_0086376 plasmid and lentivirus mimicking or inhibiting miR-5195-3p fragment were transfected into HaCaT cells, and ELISA was used to detect the expression of inflammatory factors in the culture supernatant of HaCaT cells after overexpression or interference of circ_0086376 cultured alone or with P.acne. The luciferase reporter assay was conducted to verify the targeting relationship between circ_0086376 and miR-5195-3p. Additionally, ELISA was used to detect the effects of overexpression/interference of circ_0086376 and mimic/inhibition of miR-5195-3p on the expression of inflammatory factors in the culture supernatant of HaCaT cells co-cultured with P.acne. Results:After co-culture with P.acne, the levels of inflammatory factors in the culture supernatant were significantly higher than those in the HaCaT cells cultured alone, including Interleukins (IL) -1β ([355.80 ± 23.20] vs. [260.50 ± 16.58] pg/ml, t = 5.79, P < 0.01), IL-6 ([38.04 ± 2.69] vs. [14.33 ± 0.75] pg/ml, t = 14.65, P < 0.01), IL-12 ([10.87 ± 0.78] vs. [6.52 ± 0.77] pg/ml, t = 6.89, P < 0.01), IL-18 ([222.60 ± 21.07] vs. [146.10 ± 9.14] pg/ml, t = 5.77, P < 0.01), and Tumor necrosis factor (TNF) -α ([50.39 ± 1.29] vs. [20.46 ± 0.83] pg/ml, t = 33.83, P < 0.01). The expression levels of IL-1β, IL-6, IL-12, IL-18 and TNF-α in the culture supernatant of HaCaT cells in the circ-empty vector + P.acne group were higher than those in the circ-empty vector overexpression group. The levels of inflammatory factors mentioned above in circ-overexpression + P.acne group were lower than those in circ-overexpression empty vector + P.acne group ( P < 0.01). The expression levels of inflammatory factors mentioned above in the culture supernatant of HaCaT cells in the interference circ-empty vector + P.acne group were higher than those in the interference circ-empty vector group. Compared with the interference circ-empty vector + P.acne group, the expression of circ in the interference circ + P.acne group was higher ( P < 0.01). Luciferase reporter assay confirmed that circ_0086376 could bind to miR-5195-3p. The expression levels of IL-1β, IL-6, IL-12, IL-18 and TNF-α in the circ-overexpression group were lower than those in the empty vector group ( P < 0.05), and the levels of these inflammatory factors in the circ-overexpression + miR mimic group were higher than those in the circ overexpression group ( P < 0.05). The expression levels of inflammatory factors in the interference circ group were higher than those in the empty vector group, levels of inflammatory factors in the circ + miR-inhibiting group were lower than those in the circ interference group ( P < 0.05) . Conclusion:Circ_0086376 can inhibit the expression of acne-related inflammatory factors by targeting and downregulating miR-5195-3p.
3.Role of microRNAs in inflammatory skin diseases
Ruixian YE ; Rujun XUE ; Jingyao LIANG ; Huanyan LIU ; Qiying XIONG ; Liangjiao DONG ; Xibao ZHANG
Chinese Journal of Dermatology 2021;54(2):178-182
MicroRNAs (miRNAs) are a class of non-coding RNA molecules that regulate gene expression after transcription and participate in various pathophysiological processes in the skin. In recent years, it has been reported that changes in miRNA expression profiles are related to some inflammatory skin diseases. For example, miR-203, miR-146a and miR-21 are upregulated in psoriatic lesions, miR-155 and miR-146a are upregulated in atopic dermatitis lesions, miR-21, miR-223, miR-142-3p and miR142-5p are upregulated in allergic contact dermatitis lesions; however, miR-146a and miR-155 are downregulated in peripheral blood of patients with systemic lupus erythematosus, and miR-223 is downregulated in dermatomyositis lesions. This review summarizes relationships of miRNAs with the occurrence and development of some inflammatory skin diseases.
4.Atopic dermatitis in the elderly
Xibao ZHANG ; Liangjiao DONG ; Ruixian YE ; Huanyan LIU ; Qiying XIONG
Chinese Journal of Dermatology 2021;54(5):447-451
In recent years, the category of atopic dermatitis (AD) has been updated in domestic and foreign guidelines, and elderly AD has been added as a subtype. The pathogenesis of elderly AD is related to heredity, skin barrier dysfunction, immune dysregulation and lifestyle. Most elderly AD patients have atypical clinical symptoms, and misdiagnosis is very common. To fully understand the pathogenesis and clinical characteristics of elderly AD, and to formulate individualized diagnosis and treatment plans based on clinical characteristics, are particularly important for improving the quality of life of patients and reducing the burden of the disease.
5.The effect of a motor relearning programme combined acupuncture on muscle tension and motor function recovery after cerebral infarction
Xu ZHANG ; Qihui LI ; Yingying LI ; Chuanbang CHEN ; Junyan LU ; Beilei ZHU ; Ruixian ZHENG ; Chun LIAN ; Tianshen YE
Chinese Journal of Physical Medicine and Rehabilitation 2012;34(4):279-282
Objective To observe the effects of a motor relearning programme (MRP) combined with different early acupuncture interventions on muscle tension and motor function recovery after cerebral infarction.MethodsA total of 90 patients with cerebral infarction who met the inclusion criteria were divided into three groups at random:a YANGMING meridian acupuncture and MRP group ( group A),an anti-spasm acupuncture and MRP group ( group B),and an MRP group ( group C ).All of the patients in all three groups were treated with routine medication.The National Institute of Health stroke scale (NIHSS),the composite spasticity scale (CSS),Fugl-Meyer assessment (FMA),the Fugl-Meyer balance scale (FM-B) and the modified Barthel index (MBI) were used to measure performance before treatment and after 4 weeks of treatment.Another comparison was intra-group between before and after treatment. ResultsThere were significant differences in the assessment results in all of the groups after treatment compared with those before treatment.After treatment,group B was superior to group C only in terms of NIHSS scores.There was no significant NIHSS score difference between groups A and C.The FMA,CSS and MBI results revealed significant differences among all three groups,with the scores of group A consistently the highest.The average FMA score in group B was significantly higher than in group C but there was no statistically significant difference in FM-B scores among the three groups. ConclusionMRP therapy combined with early acupuncture intervention can improve motor function and muscular tension after cerebral infarction.Anti-spasm acupuncture can improve motor function and control muscular tension effectively at the same time,making it beneficial for MRP training.
6.Effect of lymphocytes modified by brain-derived neurotrophic factor gene on the proliferation and the H_2O_2-induced apoptosis of PC12 cell
Xiaoqing TANG ; Jianqiang FENG ; Ruixian GUO ; Pingyang LI ; Meiong YE
Chinese Pharmacological Bulletin 2003;0(11):-
AIM To study the expression of brain-derived neurotrophic factor (BDNF) in lymphocytes modified by BDNF gene and its effect on the proliferation and the H 2O 2-induced apoptosis of PC12 cells for the transfer of ex vivo therapeutic gene into central nervous system (CNS) tissue with atraumatic means. METHODS Recombined BDNF cDNA plasmid was transfected by LipofectAMINE into the packing cell line PA317 and G418-resistant clones with highest titer was selected. Rat lymphocytes were infected repeatedly with virus supernatant. The expression of BDNF in rat lymphocytes was assayed by FCM and immunohistochemistry. The proliferation of PC12 cells was evaluated by MTT assay. H 2O 2-induced apoptosis of PC12 cells was determined by PI stain flow cytometry. RESULTS BDNF expressed in rat lymphocytes genetically modified and the concentration of BDNF in the conditioned medium of engineered lymphocytes had a linear correlation with the proliferation of the PC12 cells. The supernatant of lymphocytes modified by BDNF gene decreased the apoptosis of PC12 cells induced by H 2O 2. CONCLUSION Rat lymphocytes genetically modified can express and secret active BDNF in vitro.
7.The adaptive cytoprotection of H_2O_2 preconditioning against oxidative-stress damage in PC12 cell
Xiaoqing TANG ; Jianqiang FENG ; Pingyang LI ; Ruixian GUO ; Meihong YE ;
Chinese Pharmacological Bulletin 2003;0(10):-
AIM To study the adaptive cytoprotection of H 2O 2 preconditioning against oxidative stress damage in PC12 cell and the relationships between brain derived neurotrophic factor (BDNF) and the adaptive cytoprotection of H 2O 2 preconditioning. METHODS The viability of PC12 cells was evaluated by MTT assay. Apoptosis was detected by PI stain flow cytometer. Expression of BDNF was analyzed by immuo flow cytometer. RESULTS After H 2O 2 preconditioning, the survival of PC12 cells exposure to H 2O 2 (20~60 ?mol?L -1 ) was increased and the apoptosis of PC12 cells induced by H 2O 2 (20 or 30 ?mol?L -1 ) was inhibited and the expression of BDNF in PC12 cells was enhanced. CONCLUSION H 2O 2 preconditioning is protective against the damage of PC12 cells induced by H 2O 2 and its mechanisms may involve up modulation of the expression of BDNF.

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