AIM: To investigate the effects of transient receptor potential channel 6 (TRPC6) on the proliferation of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS) induced by IL-1β.METHODS: The mRNA expression of TRPC6 in synovial tissues from RA or OA patients was studied by RT-qPCR.RA-FLS were cultured by enzyme digestion and tissue adhesion methods.The method of flow cytometry was applied to identify the RA-FLS.RA-FLS were treated with different concentrations (0, 0.25, 0.5, 1, 2, 4, 8 and 16 μg/L) of IL-1β for 36 h.The cell viability was examined by CCK-8 assay.RA-FLS were incubated with IL-1β (16 μg/L) for different time (12, 24, 36, 48, 60 and 72 h), and the cell viability was measured by CCK-8 assay.The interference efficiency of TRPC6-siRNA was determined by RT-qPCR and Western blotting.After incubation in the presence or absence of IL-1β medium, the cell viability, the percentage of EdU-positive cells and the percentage of (G2/M+S) phase were measured by CCK-8 assay, EdU labeling assay and flow cytometry, respectively.RESULTS: The mRNA expression of TRPC6 was found in synovial tissue with higher levels in RA patients than that in OA patients.TRPC6-siRNA significantly decreased the mRNA and protein expression of TRPC6 (P<0.05).When RA-FLS were treated with IL-1β, the proliferation of RA-FLS was increased (P<0.05).The differences of the cell viability, the percentage of EdU-positive cells and the (G2/M+S) phase percentage between TRPC6-siRNA group and blank control group or NC-siRNA group were significant, in the presence of IL-1β (P<0.05).However, they were not significant in the absence of IL-1β.CONCLUSION: TRPC6 is involved in the proliferation of RA-FLS induced by IL-1β.Silencing of TRPC6 gene inhibits the growth of RA-FLS induced by IL-1β.

BACKGROUND:Triggering receptor expressed on myeloid cel s 2 (TREM-2) is highly expressed throughout the synovial tissue in active rheumatoid arthritis patients, but the role of TREM-2 in the pathogenesis of rheumatoid arthritis stil remains unclear.
OBJECTIVE:To explore the TREM-2 expression in the synovial tissue of col agen type II-induced arthritis rats.
METHODS:The col agen-induced arthritis models were established in rats. The activity indicators and pathological changes of arthritis synovial were dynamical y observed. The mRNA levels of TREM-2, tumor necrosis factor-α, interleukin-1β, and interleukin-10 were detected in synovial tissue of rats by RT-PCR. The protein expression and location of TREM-2 were measured with western blot assay and immunohistochemistry, respectively.
RESULTS AND CONCLUSION:At day 13 after immunization, the paws of model rats appeared red and swel ing, the arthritis index scores were increased (P<0.01). At day 19-25 after immunization, the inflammation reached the peak. Hematoxylin-eosin staining showed that, the synovium of col agen-induced arthritis rats were proliferated and were infiltrated by inflammatory cel s, cartilage was destroyed. Compared with the control group, the expression of TREM-2 mRNA and protein, the mRNA levels of tumor necrosis factor-αand interleukin-1βin synovial tissue of the model rats were significantly increased (P<0.05 or P<0.01), while interleukin-10 mRNA expression was significantly decreased (P<0.05). Experimental findings indicate that, TREM-2 is a crucial inflammatory regulator and the increasing expression of TREM-2 plays an important role in the pathogenesis of col agen-induced arthritis.

Objective To observe humoral and cellular immune responses induced in mice by a TSOL18 recombinant Mycobacterium smegmatis (MS) vaccine of Taenia solium.Methods Sixty specific pathogen-free (SPF) Kunming mice (18-22 g,half male and half female) were divided into 3 groups by random number table method,20 mice in each group.One group was administered with recombinant MS-TSOL18 vaccine,one group was administered with MS as control,and one group was administered phosphate buffer saline (PBS) as control.Kunming mice were vaccinated once every two weeks for 2 times through intragastric administration.At 0,2,4,6,and 8 weeks after immunization,blood sample was collected and serum was separated.The levels of IgG and IgG2a in serum were detected using enzyme linked immunosorbent assay (ELISA).The spleen was separated to prepare spleen suspension.The proliferation of spleen lymphocyte with the stimulation of a specific antigen was detected by CCK-8.The levels of interleukin (IL)-2 and IL-4 in spleen cell culture supernatant with the stimulation of a specific antigen were detected by ELISA.Results Compared with MS control group (IgG:0.160 ± 0.019,0.187 ± 0.038,0.193 ± 0.050,0.170 ± 0.005;IgG2a:0.213 ± 0.010,0.198 ± 0.012) and PBS control group (IgG:0.159 ± 0.015,0.184 ± 0.029,0.191 ± 0.025,0.165 ± 0.018;IgG2a:0.198 ± 0.032,0.178 ± 0.025),the levels of specific IgG (0.310 ± 0.034,0.391 ± 0.029,0.443 ± 0.030,0.373 ± 0.021) in recombinant MS-TSOL18 vaccine group increased at 2,4,6,and 8 weeks after immunization (P ＜ 0.05),the levels of specific IgG2a (0.446 ± 0.056,0.339 ± 0.026) in recombinant MS-TSOL18 vaccine group increased at 6 and 8 weeks after immunization (P ＜ 0.05),and reached the highest level by the 6th week.After antigen stimulation,compared with MS and PBS control groups,the levels of spleen lymphocyte proliferation increased at 2,4,6,and 8 weeks after immunization (P ＜ 0.05),and reached the highest level by the 6th week.After antigen stimulation,compared with MS and PBS control groups,the levels of IL-2 and IL-4 in spleen lymphocyte culture supernatant increased at 2,4,6,and 8 weeks after immunization (P ＜ 0.05),and reached the highest level by the 6th and 4th weeks,respectively.Conclusion The recombinant MS-TSOL18 vaccine of Taenia solium might induce mice to produce humoral and cellular immune responses.