Moxibustion regulates T-regulatory/T-helper 17 cell balance by modulating the microRNA-221/suppressor of cytokine signaling 3 axis in a mouse model of rheumatoid arthritis / 中西医结合学报

OBJECTIVE@#Rheumatoid arthritis (RA) progression is associated with the balance of T-regulatory (Treg) and T-helper 17 (Th17) cells, while the role of microRNAs (miRs) in regulating Treg/Th17 cell balance has not been clarified. This study aimed to assess whether moxibustion could regulate Treg/Th17 cell balance by modulating the miR-221/suppressor of cytokine signaling 3 (SOCS3) axis in the RA mouse model.@*METHODS@#A mouse model of collagen-induced arthritis (CIA) was established in male DBA/1J mice. Twenty-two days after CIA induction, the mice received daily treatment with moxibustion for 12 times. Pathological scores were assessed according to the levels of synovial hyperplasia. The expression levels of cytokines interleukin (IL)-1β, IL-6, tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), IL-17 and IL-10 were analyzed in serum by enzyme-linked immunosorbent assay. The cluster of differentiation 4 (CD4+) splenocytes was analyzed by fluorescence-activated cell sorting. The expression levels of RA-related miRs and target genes were subsequently detected, and the target of miR-221 was confirmed by the dual-luciferase reporter assay.@*RESULTS@#It was revealed that moxibustion treatment decreased the pathological scores and downregulated the expression levels of IL-1β, IL-6, TNF-α, IFN-γ and IL-17, while upregulated the expression level of IL-10. The Treg/Th17 cell balance was regulated by moxibustion treatment. The expression level of miR-221 was suppressed by moxibustion treatment. Furthermore, SOCS3 was found as the direct target of miR-221, which mediated the function of moxibustion by regulating the Treg/Th17 cell balance.@*CONCLUSION@#Moxibustion therapy regulated the Treg/Th17 cell balance by modulating the miR-221/SOCS3 axis in the RA mouse model.

Effect of Buyi Pishen acupuncture on inflammatory factor and cartilage matrix in adjuvant arthritis rats / 中国针灸

OBJECTIVE@#To observe the effect of Buyi Pishen acupuncture (acupuncture for invigorating spleen and kidney) on inflammatory factor and synovial cartilage matrix in adjuvant arthritis (AA) rats, and to explore the mechanism of acupuncture for rheumatoid arthritis (RA).@*METHODS@#A total of 60 clean male Wistar rats were randomized into a normal group, a model group, a tripterygium wilfordii polyglycoside tablet (TWP) group and an acupuncture group, 15 rats in each group. Rats in the model group, the TWP group and the acupuncture group received intradermal injection of Freund's complete adjuvant (FCA) at right hind foot pad to induce the AA model. TWP suspension of 8 mg/kg was given by gavage in the TWP group. Acupuncture was applied at "Shenshu" (BL 23), "Pishu" (BL 20) and right "Housanli" (ST 36), "Sanyinjiao" (SP 6), "Yanglingquan" (GB 34) in the acupuncture group, 15 min a time, once a day. The intervention was given 15 days in both TWP group and acupuncture group. The foot-pad swelling degree before modeling, before and after intervention and the arthritis index (AI) score before and after intervention were calculated; the serum levels of interleukin (IL)-1β, IL-4, IL-10 and tumor necrosis factor-α (TNF-α) were detected by ELISA method; the ultrastructure and histomorphological changes of synovium issue were observed by transmission electron microscope and HE staining; the positive expression of matrix metalloproteinase (MMP)-3 and MMP-9 in synovium issue was detected by immunohistochemistry method.@*RESULTS@#Before intervention, foot-pad swelling degree of the model group, the TWP group and the acupuncture group was increased compared with the normal group (P<0.01). After intervention, foot-pad swelling degree and AI score were increased compared with the normal group (P<0.01), foot-pad swelling degree and AI scores in the TWP group and the acupuncture group were lower than the model group (P<0.05), and those in the acupuncture group were decreased compared with the TWP group (P<0.05). The model group exhibited unclear nuclear membrane of synovial cells, chromatin pyknosis, massive inflammatory cell infiltration and hyperplasia in synovial tissue; the TWP group and the acupuncture group exhibited clear and smooth nuclear membrane of synovial cells, inapparent chromatin pyknosis, less inflammatory cell infiltration and hyperplasia in synovial tissue, the acupuncture group exhibited less matrix destruction as well. Compared with the normal group, serum levels of IL-1β and TNF-α and positive expression of MMP-3 and MMP-9 in synovium issue were increased (P<0.01), while serum levels of IL-4 and IL-10 were decreased (P<0.01) in the model group. Compared with the model group, serum levels of IL-1β and TNF-α and positive expression of MMP-3 and MMP-9 in synovium issue were decreased (P<0.05, P<0.01), while serum levels of IL-4 and IL-10 were increased (P<0.05) in the TWP group and the acupuncture group; compared with the TWP group, serum level of TNF-α and positive expression of MMP-3 and MMP-9 in synovium issue were decreased (P<0.05), while serum levels of IL-4 and IL-10 were increased (P<0.05) in the acupuncture group.@*CONCLUSION@#Buyi Pishen acupuncture can effectively improve the injury of articular cartilage in AA rats, its mechanism maybe related to reducing the inflammatory reaction in synovium and inhibiting the degradation of articular cartilage matrix.

Effect of moxibustion at "Zusanli" (ST 36) and "Shenshu" (BL 23) on intestinal flora in adjuvant arthritis rats / 中国针灸

OBJECTIVE@#To observe the effect of moxibustion at "Zusanli" (ST 36) and "Shenshu" (BL 23) on inflammatory factors and intestinal flora in the rats with adjuvant arthritis.@*METHODS@#A total of 36 Wistar rats were randomized into a normal group, a model group and a moxibustion group, 12 rats in each one. In the model group and the moxibustion group, the adjuvant arthritis model was established by a compound method, including the environmental factors, i.e. wind, cold and damp, and Freund's complete adjuvant. In the moxibustion group, moxibustion intervention was exerted at "Zusanli" (ST 36) and "Shenshu" (BL 23), for 20 min at each acupoint, once daily, consecutively for 21 days. The paw swelling degree and arthritis index (AI) score were observed before and after intervention in the rats of each group. Using real-time fluorescence quantitative PCR method (real-time PCR) and Western blot method, the mRNA and protein expressions of inflammatory factors of colon tissue, i.e. interleukin (IL) 1β, tumor necrosis factor-α (TNF-α), IL-6, were detected after intervention in the rats of each group. The intestinal flora was detected with 16SrRNA sequencing technology after intervention in the rats of each group.@*RESULTS@#Compared with the normal group, the paw swelling degree and AI score were increased in the rats of the model group (@*CONCLUSION@#Moxibustion at "Zusanli" (ST 36) and "Shenshu" (BL 23) relieves the joint symptoms of adjuvant arthritis rats and inhibits the expressions of inflammatory factors, which is probably related to the regulation of the structure of intestinal flora.

Therapeutic effect of gene silencing peptidyl arginine deaminase 4 on pulmonary interstitial lesions induced by collagen-induced arthritis mice / 北京大学学报(医学版)

OBJECTIVE@#To investigate the therapeutic effect of gene silencing peptidyl arginine deaminase 4 (PAD4) on pulmonary interstitial lesions induced by collagen-induced arthritis (CIA) mice, and possible mechanisms.@*METHODS@#A CIA mouse model was established in DBA/1 mice, followed by a tail vein injection of the virus solution prepared by the PAD4-siRNA expression vector once a week for 8 times. The mice were sacrificed at the end of the experiment. The expression of PAD4 mRNA in lungs was detected by real-time quantitative PCR (qRT-PCR). The expression of PAD4 protein was detected by tissue immunohistochemistry. Cell culture was performed by spleen tissue. Flow cytometry changes in the ratio of Tfh cells to Tfr cells were examined; lung staining was performed in the lungs to observe changes in lung pathology.@*RESULTS@#(1) Compared with the blank group, the expression of PAD4 mRNA in the lung tissue of the model group increased, the difference was statistically significant (P < 0.05). PAD4 mRNA in the lung tissue of the CIA mice after PAD4-siRNA treatment. The expression level was significantly lower than that of the model group and the negative control group, and the difference was statistically significant (P < 0.05). (2) Red fluorescence was less in the lung tissue of the blank group, while more red fluorescence was observed in the inflammatory cell infiltration area and trachea around the lung tissue of the model group and the negative control group, and the red fluorescence of the three groups after PAD4-siRNA treatment was significantly reduced; (3) Compared with the blank group, the proportion of Tfh cells in the model group increased, the difference was statistically significant (P < 0.05), the proportion of Tfh cells in spleen cells of the CIA mice after PAD4-siRNA treatment was significantly lower than that of the model group and the negative control group, the difference was statistically significant (P < 0.05); compared with the blank group, in the mouse spleen cells in the model group the proportion of Tfr cells was slightly decreased, but the difference was not statistically signifi-cant. The proportion of Tfr cells in the spleen cells of the mice increased after PAD4-siRNA treatment, but the difference was statistically significant only in the PAD4-siRNA2 group compared with the model group and the negative control group (P < 0.05); (4) The proportion of Tfh/Tfr in the spleen cells of the model group was increased, compared with the blank group, the difference was statistically significant (P < 0.05); the ratio of Tfh/Tfr in the three groups after PAD4-siRNA treatment all decreased, the difference was statistically significant (P < 0.05); (5) Compared with the blank group, the alveolar wall of the lung tissue of the model group was thickened, the inflammatory cell infiltration was increased, and the lung tissue destruction and inflammatory infiltration of the CIA mice were decreased after PAD4-siRNA treatment. The degree of reduction was reduced.@*CONCLUSION@#Gene silencing of PAD4 can reduce the proportion of Tfh cells, increase the proportion of Tfr cells, reverse the proportion of Tfh/Tfr, and reduce the degree of interstitial lesions and inflammatory infiltration of lung tissue.

Efeito do exercício aeróbico em modelo experimental de artrite / Effect of aerobic exercise on an experimental model of arthritis

INTRODUÇÃO: A artrite reumatoide é uma doença inflamatória sistêmica autoimune que acomete preferencialmente as articulações, mas também outros tecidos, como o músculo esquelético. A perda de massa muscular determina uma grande repercussão na funcionalidade e qualidade de vida desses pacientes e o exercício físico surge como uma alternativa terapêutica para esse acometimento. OBJETIVO: Avaliar o efeito do exercício físico aeróbico moderado sobre a perda muscular em artrite induzida por colágeno (CIA). MÉTODOS: Esse é um estudo-piloto em que CIA foi induzida em camundongos machos DBA1/J divididos em dois grupos: (i) animais com exercício (EXE, n=5), (ii) animais sem exercício (semEXE, n=4). Foram avaliados o escore clínico, o edema da pata traseira, o peso do animal e a locomoção espontânea periodicamente. Após a morte, a histopatologia da articulação tibiotarsal e a área da miofibra dos músculos gastrocnêmio e tibial anterior foram avaliados. Significância foi considerada se p<0,05.RESULTADOS: Não foi observada diferença significativa entre os grupos nos parâmetros de atividade da doença, peso e locomoção espontânea. Entretanto, a histopatologia da articulação demonstrou redução da erosão cartilaginosa no grupo EXE. Também se observou aumento significativo na área seccional da miofibra do grupo EXE, representando uma diferença média de 24%. CONCLUSÃO: Este é o primeiro estudo com exercício aeróbico moderado em esteira em modelo experimental de artrite. O protocolo de exercício testado não parece impactar no desenvolvimento clínico da doença, mas demonstrou benefício sobre a perda muscular consequente da artrite, reduzindo a atrofia da miofibra.

BACKGROUND: Rheumatoid arthritis (RA) is a systemic autoimmune inflammatory disease that affects primarily the joints, but also other tissues such as skeletal muscle. Muscle wasting significantly impairs the functionality and quality of life of patients with RA and physical exercise is an alternative therapy for this outcome. AIM: To evaluate the effect of moderate aerobic physical exercise on muscle loss caused by collagen-induced arthritis (CIA). METHODS: This is a pilot study in which CIA was induced in DBA/1J mice divided into two groups: (i) animals which exercised (EXE, n=5), (ii) animals which did not exercise (semEXE, n=4). Clinical score, hind paw swelling, weight, and spontaneous locomotion were evaluated periodically. After death, the histopathological score of the ankle and the myofiber area of the gastrocnemius and tibialis anterior muscles were evaluated. Significance was considered when p<0.05. RESULTS: No significant difference was observed between groups regarding clinical parameters of disease activity, animal weight, and spontaneous locomotion. However, joint histopathology demonstrated a decrease in cartilage erosion in the EXE group. There was also significant difference in the myofiber sectional area, with a 24% increase in the EXE group. CONCLUSION: This is the first interventional study with moderate aerobic exercise on a treadmill in an arthritis experimental model. The tested exercise program does not seem to have a clinical impact on the process of arthritis. However, it has a positive effect on muscle wasting caused by arthritis, demonstrated mainly by the reduction of myofiber atrophy.

Inmunopatologia de la lepra / Immunopathology of leprosy

Se comentan los estudios en convivientes íntimos de enfermos de Lepra, donde se observó una relación inversa entre grado y tiempo de exposición al Mycobacterium leprae (MI) y la magnitud de la respuesta inmune in vivo hacia el bacilo (reacción de Mitsuda); esta variable por otro lado, se halla influenciada por un importante grado de determinación genética. En un modelo murino experimental, la artritis por adyuvante, también se comprobó que el tratamiento con MI inducia una población de células esplénicas supresoras, con actividad regulatoria sobre la respuesta inmune celular. Se plantean además, las posible alteraciones inmunorregulatorias involucradas con la excesiva respuesta humoral que sirve de sustrato para la generación del episodio de eritema nudoso leproso (ENL)

Imunopatologia de la lepra / Immunopathology of leprosy

Se comentan los estudios en convivientes íntimos de enfermos de Lepra, donde se observó una relación inversa entre grado y tiempo de exposición al Mycobacterium leprae (MI) y la magnitud de la respuesta inmune in vivo hacia el bacilo (reacción de Mitsuda); esta variable por otro lado, se halla influenciada por un importante grado de determinación genética. En un modelo murino experimental, la artritis por adyuvante, también se comprobó que el tratamiento con MI inducia una población de células esplénicas supresoras, con actividad regulatoria sobre la respuesta inmune celular. Se plantean además, las posible alteraciones inmunorregulatorias involucradas con la excesiva respuesta humoral que sirve de sustrato para la generación del episodio de eritema nudoso leproso (ENL)