1.TCM pathogenesis of Sjogren's syndrome
International Journal of Traditional Chinese Medicine 2010;32(4):366-367
The main pathogenesis of Sjogren's syndrome was Yin Deficiency, dryness caused by blood stasis,dryness caused by dry poisoning, dryness caused by dampness, Qi deficiency, and incoordination of spleen and stomach,which led to damage of body fluid or transport barrier. Usually such factors were mutually intercross and interact, existing in the whole process of the disease. This is the important factor for a long course and intractable disease.
2.The Influence of Runzaoling on AQP1 and AQP5 in submaxillary gland of mice with Sjogren syndrome
Ertao JIA ; Limin LIU ; Xueming YAO ; Wukai MA
International Journal of Traditional Chinese Medicine 2011;33(4):316-318
Objective To study the influence of the Runzaoling on AQP1 and AQP5 in submaxillary gland of mice with Sjogren syndrome.Methods Sjogren syndrome mice models were setup by innducement method and divided into six groups randomly,including A group(blank group),B group(model group),C group (low dose group),D group(moderate dose group),E group(high dosage group)and F group(prednisone group).Immunohistochemical method was used to detect the of expressions of AQP1 and AQP5 of submaxillary gland in model mice.Result AQP1 expression showed:scattered flavescent particles were only found in submaxillary gland of group B and group C;AQP5 expression showed:brown-stained granules can be found in gland alveolous teleblem,lateral membrane,and basal lamina of submaxillary gland,secretory duct,and duct epithelia in B group mice;light brawn-stained granules can be found in group C mice;attenuated or disappear stained granules were found in acinus,secretory tube,and duct in D,E,and F group mice.Conclusion Runzaoling Can raise the expression of AQP5 in submaxillary gland of SS mice and increase the absorption quantity of water.
3.Effects of Ginger Phenol on IL-17 and IL-23 in Serum of Rheumatoid Arthritis Model Rats
Wukai MA ; Xueming YAO ; Ying HUANG ; Fang TANG ; Ying WANG ; Jing ZHOU
Chinese Journal of Information on Traditional Chinese Medicine 2014;(9):43-45
Objective To observe the effects of Chinese medicine ingredient ginger phenol on expressions of IL-17 and IL-23 in collagen-induced arthritis (CIA) rats;To investigate the role and mechanism of ginger phenol in CIA rats.Methods Forty female Wistar rats were randomly divided into 4 groups:blank group, model control group, ginger phenol treatment group and tripterygium glucosides control group. After CIA was established in rats successfully, ginger phenol treatment group and tripterygium glycosides control group were treated with ginger phenol and tripterygium glycosides by gavage, while the control group and model control group were fed with the same amount of saline. Four weeks after the treatment, the rats were collected serum and sacrificed. The pathological of CIA joint synovial was observed by electron microscope. The expressions of IL-17 and IL-23 in serum were detected by ELISA.Results The hyperplasia of synovial tissue and angiogenesis was reduced in gingerol phenol treatment group. Infiltration of inflammatory cells and edema was improved. IL-17 and IL-23 expressions of in serum of CIA rats were significantly higher than those of the blank group (P<0.01). Ginger phenol and tripterygium glycosides down-regulated the expressions of IL-23 and IL-17 (P<0.05,P<0.01). Ginger phenol group was better than that of tripterygium glycosides control group (P<0.05).Conclusion Chinese medicine ingredient ginger phenol can reduce expressions IL-17 and IL-23 in serum of CIA rats.
4.Effect of Jinwu Jiangu Decoction on IL-17,ACT1,TRAF6 in Synovial Cells of Rheumatoid Arthritis
Wukai MA ; Qiaoyi NING ; Dongxiao LI ; Xueming YAO ; Lei HOU ; Jiang LIANG ; Ying HUANG ; Jing ZHOU
Herald of Medicine 2017;36(6):617-621
Objective To investigate the effect of Miao medicine Jinwu Jiangu decoction containing serum freeze-dried powder on levels of IL-17,ACT1 and TRAF6 in human rheumatoid arthritis fibroblast like synoviocytes (RA-HFLS).Methods Rabbits were randomly divided into blank control group (recieving normal saline of the same volume),Jinwu Jiangu decoction high-dose,medium-dose and low-dose group (intragastrically administrated with Jinwu Jiangu decoction at doses of 14.4,4.8 and 2.4 g·kg-1,respectively),tripterygium glycosides group and prednisone group (treated with human equivalent dosage).RA-HFLS primary cell model was established in the experiment.ELISA method was used to detect effect of lyophilized powder on IL-17 secretion.Expression of ACT1,TRAF6 mRNA was detected by RT-PCR.Results Compared with the blank control gorup,IL-17 in the supernatant of each medication administration group was significantly decreased (all P<0.01),and it was decreased most significantly in Jinwu Jiangu decoction high-dose and medium-dose group.IL-17 was down-regulated more significantly in high-dose group than that in tripterygium glycosides group (P<0.01).Compared with the blank control group,TRAF6 and ACT1 mRNA expression level of each medication administration group were significantly decreased (all P<0.01),and in the high-dose group that were decreased most significantly,but not significantly different as compared with tripterygium glycosides group and prednisone group (P>0.05).Conclusion Freeze-dried powder of Jinwu Jiangu decoction can decrease the secretion of IL-17 and down-regulate expression of ACT1,TRAF6 with RA-HFLS.
5.Influence of Fuzheng Toudu Qudu Recipe on IL-2 and sIL-2R Contents in Patients with Minimal Residual Disease of Myelogenous Leukemia During Dendritic Cells Derived from CD34+Cells
Jianghui QIU ; Liming HUANG ; Guojing ZHAO ; Yiqing SONG ; Jian LIU ; Wukai MA ; Yuhong YAO ; Xiujun LI ; Zhiyu TANG ; Lei WANG
Journal of Guangzhou University of Traditional Chinese Medicine 2014;(5):762-767
Objective To study the influence of serum containing Fuzheng Toudu Qudu Recipe, a Chinese formula with the actions of supporting healthy qi to expel and remove toxicity, on serum levels of interleukin 2 (IL-2) and soluble interleukin 2 receptor (sIL-2R) at different stages of CD34+derived dendritic cells (DC) of patients with minimal residual disease of myelogenous leukemia ( MRD-L) , and to explore the biological mechanism of Fuzheng Toudu Qudu Recipe in promoting CD34+ to transform into DC in MRD-L patients. Methods Bone marrow mononuclear cells ( BMMC) were separated from the bone marrow of acute myeloid leukemia patients at complete remission stage by using Ficoll centrifugation. CD34+ cells were isolated by using immuno-magnetic mircobeads method, and then were cultured with various concentrations of Chinese medicine medicated serum and cytokines in vitro for the induction of DC. The morphologic characteristics of DC were observed with the inverted phase contrast microscope, and the expression levels of DC surface molecules such as CD83, CD80, CD86, CD1a and HLA-DR were detected by using flow cytometry. On culturing day 0, 6 and 9, serum levels of IL-2 and sIL-2R of each group were measured by enzyme-linked immunosorbent assay ( ELISA). Results ( 1) Chinese medicine medicated serum combined with cytokines was effective on promoting CD34+ to differentiate into DC with typical morphology, and inducing DC to have high expression of CD80, CD83, CD86 and HLA-DR, which differed from those in fetal calf serum (FCS) group and blank rabbit serum group (P<0.01). Middle- and low-dose combination groups increased expression of CD1a, which differed from high-dose combination group and cytokines group ( P<0.01). ( 2) Content of IL-2 in combination groups was higher than that in blank rabbit serum group on culturing day 0. In the combination groups, IL-2 was higher on culturing day 6 and 9 than that on culturing day 0. Middle and high-dose combination groups had higher IL-2 content on culturing day 9 than on culturing day 6 ( P<0.05 or P<0.01). At the same time point, combination groups had higher IL-2 content than the blank rabbit serum group (P<0.05 or P<0.01). (3) Content of sIL-2R in the combination groups was lower than that in blank rabbit serum group on culturing day 0. In the combination groups, sIL-2R was lower on culturing day 9 than that on culturing day 0 and 6 ( P<0.01) . High-dose combination group had lower sIL-2R content on culturing day 6 than that on culturing day 0 (P<0.05), and the difference of sIL-2R in other groups was insignicant between on culturing day 6 and on culturing day 0 ( P>0.05) . At the same time point, combination groups had lower IL-2 content than the blank rabbit serum group (P<0.05 or P<0.01). Conclusion Fuzheng Toudu Qudu Recipe is effective on increasing serum content of IL-2 and reducing sIL-2R content, and the changes of cytokine contents are more obvious along with the maturity of DC, which indicates that the recipe plays positive effect in the process of promoting CD34+cells to differentiate into DC.
6.The role of IL-17A in the pathogenesis of primary Sj?gren′s syndrome
Xi MA ; Fangqin HUANG ; Zhaowei HUANG ; Ping ZENG ; Wukai MA
Chinese Journal of Rheumatology 2023;27(10):687-692
Objective:To investigate the role of IL-17A in the regulation of inflammatory factors and autophagy genes of PBMCs in pSS patients.Methods:Thirty patients fulfilled the diagnosis of pSS were selected, 20 mL of peripheral blood was drawn, PBMCs were isolated and divided into the PBMCs group, IL-17A stimulant group and IL-17A inhibitor group. After warm incubation 48 h of immunofluorescence was applied to detect microtubule-associated protein l light chain 3 (LC3), and the ELISA method was used to detect the expression of the inflammatory factors IL-4, IFN-γ, IL-13 expression. Real-time fluorescence polymerase chain reaction (qRT-PCR) was used to detect the expression of autophagy-inducing genes Ambra-1, Bif-1 and apoptosis genes Bcl-2 and Bcl-XL mRNA, and immunoprotein blotting was used to detect the expression of Beclin1 and LC3 protein. ANOVA was used to compare the differences between groups, and t-test was used for two-by-two comparisons. Results:The immunofluorescence results showed a significant increase in LC3 autophagic vesicles in the IL-17A inhibited group compared with the IL-17A stimulator group. The ELISA results showed that, compared with the PBMCs group [IL-4: (13.39±0.32) pg/ml, IFN-γ: (14.4±0.4) pg/ml, and IL-13: (854±36) pg/ml], IL-4 secretion in the IL-17A stimulated group (11.54±0.30) was decreased ( t=12.83, P=0.024), IFN-γ and IL-13 secretion [(17.6±0.4), (908±51) pg/ml] were increased ( t=19.35, P=0.033; t=2.55, P=0.020); compared with IL-17A inhibitor group [IL-4: (15.65±0.26) pg/ml, IFN-γ: (13.6±0.3) pg/ml, and IL-13: (792±57) pg/ml]. Compared with the IL-17A stimulator group, IL-4 secretion was decreased ( t=21.31, P=0.006), and IFN-γ and IL-13 expression was increased ( t=17.34, P=0.015; t=5.14, P=0.007). The PCR results showed that, compared with Ambra-1, Bif-1, Bcl-2, and Bcl-XL mRNA expression (5.61±0.33, 5.04±0.60, 1.28±0.09, 1.56±0.03) in the PBMCs group, Ambra-1, Bif-1 mRNA in the IL-17A-stimulated group expression (3.76±0.24, 4.68±0.41) were down-regulated ( t=14.30, P=0.007; t=15.02, P=0.012), and Ambra-1, Bif-1 mRNA expression (7.91±1.17, 9.30±0.25) were increased in the IL-17A inhibition group, ( t=13.59, P=0.025; t=11.54, P=0.031), anti-apoptotic proteins Bcl-2, Bcl-XL mRNA expression (1.75±0.06, 2.43±0.16) was up-regulated in IL-17A stimulated group ( t=19.92, P=0.006; t=21.04, P=0.007) were up-regulated and Bcl-2, Bcl-XL mRNA expression (0.48±0.03, 0.83±0.10) were down-regulated in the IL-17A inhibition group ( t=29.44, P=0.027; t=16.31, P=0.023). The results of protein blotting assay showed that, Beclin-1 and LC3 protein expression (0.51±0.10, 0.559±0.010) were decreased in IL-17A stimulated group compared with Beclin-1, LC3 protein expression (0.72±0.09, 0.635±0.017) in PBMCs group ( t=14.38, P=0.034; t=17.99, P=0.014); BecLin-1 and LC3 protein expression (0.83±0.11, 0.737±0.025) increased in the IL-17A inhibition group ( t=9.72, P=0.027; t=22.35, P=0.007). Conclusion:IL-17A plays a role in pSS by regulating the expression of inflammatory factors IL-4, IFN-γ, IL-13 and autophagy related genes Beclin1 and LC3.
7.The emerging role of blood lipoprotein levels in osteoporosis of postmenopausal women with rheumatoid arthritis
Zong JIANG ; Xiaoling YAO ; Fang TANG ; Wukai MA
Chinese Journal of Rheumatology 2023;27(7):446-451
Objective:To retrospectively analyze the correlation between blood lipoprotein levels and the risk of osteoporosis (OP) development in postmenopausal patients with RA and its influencing factors.Methods:Patients hospitalized with a definite diagnosis of RA from July 2017 to May 2020 were retrospectively collected and analyzed by bone mineral density (BMD) in subgroups, using correlation analysis, di-chotomous Logistic regression to quantify independent associations between laboratory test results and out-comes, and restrictive cubic spline (RCS) to fit the relationship of OP risk occurrence.Results:Six hundred and sixty-six eligible RA patients were included according to inclusion criteria, including 253 RA-OP and 413 RA-non-OP patients. After exclusion of relevant influencing factors by comparing demographic characteristics, a significant correlation was found between blood HDL-C ( r=-0.11, P=0.006) LDL-C ( r=0.12, P=0.003) levels and RA-OP( P<0.05), and dichotomous Logistic regression showed that as BMI ( OR(95% CI)=0.81(0.77, 0.86), P<0.001], calcium [ OR(95% CI)=0.24(0.10, 0.63), P<0.001], HDL-C[ OR(95% CI)=0.38(0.22, 0.66), P<0.001] increased, the risk of developing OP in RA patients decreased. In contrast, the risk of developing OP increased with increasing age [ OR(95% CI)=1.10(1.07, 1.21), P<0.001), disease duration [ OR(95% CI)=1.00(1.00, 1.00), P=0.020], and LDL-C[ OR(95% CI)=1.71(1.38, 2.12), P<0.001]. Conclusion:Blood HDL-C and LDL-C levels are significantly correlated with the development of RA-OP, and can be used as predictors of OP development and good indicators for disease monitoring in RA patients.
8.Clinical Effect of Runzaoling Combined with Iguratimod on Primary Sjögren's Syndrome
Linru LUO ; Xi MA ; Ping ZENG ; Lei HOU ; Wukai MA
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(2):127-133
ObjectiveTo investigate the clinical efficacy and safety of iguratimod combined with the Chinese medicine Runzaoling in the treatment of primary Sjögren's syndrome (pSS). MethodSeventy-two patients treated in the Department of Rheumatology and Immunology of the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine(TCM) from January 2021 to June 2022 who met the Western medical diagnosis of pSS and had the TCM syndrome of Yin deficiency and heat toxin syndrome were randomly assigned into an observation group and a control group, with 36 patients in each group. The observation group was treated with iguratimod combined with Runzaoling, and the control group was treated with iguratimod. The treatment in both groups lasted for 12 weeks. The clinical symptoms, EULAR Sjogren's syndrome patient reported index (ESSPRI), EULAR Sjögren's syndrome disease activity index (ESSDAI), erythrocyte sedimentation Rate (ESR), C-reactive protein (CRP), immunoglobulin (IgG), Schirmer score, and saliva flow of the two groups were determined before and after treatment. Furthermore, the incidence of adverse reactions was compared between the two groups. ResultThe total response rate in the observation group was 75.0% (27 patients with response and 9 patients with no response), which was higher than that (61.11%, 22 patients with response and 14 patients without response) in the control group (P<0.05). After treatment, the ESSPRI, ESSDAI, and TCM syndrome scores in both groups decreased and the decreases were more obvious in the observation group than in the control group (P<0.05). The treatment in both groups recovered the ESR, CRP, IgG, Schirmer score, and saliva flow (P<0.05). Moreover, the observation outperformed the control group in terms of the ESR, CRP, IgG, and saliva flow (P<0.05) and had no significant difference in the Schirmer score compared with the control group. During the treatment period, 2 patients in the observation group had nausea, and 1 patient had an abnormal liver function, which were relieved after symptomatic treatment and did not affect the treatment. In the control group, 1 patient withdrew from the study due to rashes and showed no special discomfort in the follow-up 4 weeks, and 1 patient had nausea, which was relieved after symptomatic treatment. ConclusionIguratimod combined with Runzaoling has good clinical efficacy and safety in the treatment of pSS.
9.The expression of System Xc-/ GSH / GPX4 ferroptosis pathway in peripheral blood mononuclear cells of rheumatoid arthritis patients and its effect on the secretion of inflammatory factors
Can Liu ; Wukai Ma ; Changming Chen ; Yang An ; Zong Jiang ; Hai Huang
Acta Universitatis Medicinalis Anhui 2024;59(1):64-70
Objective :
To investigate the expression of genes and proteins in the peripheral blood mononuclear cell ( PBMC) cystine / glutamate antiporter system (System Xc-) / glutathione ( GSH) / glutathione peroxidase 4 ( GPX4) ferroptosis pathway and its influence on inflammatory factors in patients with rheumatoid arthritis ( RA) .
Methods :
30 patients with RA and 30 healthy participants were enrolled.PBMCs were isolated using Ficoll-hypaque density gradient centrifugation.The cells were categorized into the healthy control,RA,ferroptosis inhibitor,ferroptosis in- ducer group.The cell viability was checked using the cell counting kit-8 ( CCK-8) method.Intracellular Fe2 + rela- tive fluorescence intensity and reactive oxygen species (ROS) levels were detected using the FerroOrange and Di- hydroethidium (DHE) fluorescent probes,respectively.Western blot and real-time quantitative PCR (qPCR) de- tected the expression of nuclear factor erythroid 2-related factor 2 ( Nrf2 ) ,solute carrier family 7 member 11 (SLC7A11) ,GPX4 proteins and mRNA.And the flow cytometry quantified the levels of tumor necrosis factor α (TNF-α) ,Interleukin (IL) -1,and IL-6 in the supernatant of each cell group.
Results :
Compared to the healthy control group,the RA group showed a significantly increased Fe2 + concentration and elevated ROS levels,reduced expression of Nrf2,SLC7A11 and GPX4 proteins and mRNA,and increased contents of TNF-α , IL-1 and IL-6 in PBMC supernatant,and the differences were statistically significant.The concentration of Fe2 + and ROS levels in the inhibitor group were lower than those in the RA group,the proteins expressions of Nrf2,SLC7A11 and GPX4 increased,the mRNA expressions of SLC7A11 and GPX4 increased,the content of IL-6 in the PBMC supernatant decreased but the content of TNF-α increased,and the differences were statistically significant.In contrast,the in- ducer group,when compared to the RA group,displayed increased ROS levels,reduced expression of SLC7A11 protein and mRNA and decreased expression of Nrf2 protein,and the contents of TNF-α and IL-1 in the PBMC su- pernatant increased,but the expression of GPX4 protein increased ,and the differences were statistically signifi- cant.The inducer group,compared to the RA group,showed increased cell viability,and the difference was statis-
tically significant (P<0. 000 1) .
Conclusion
The presence of ferroptosis in PBMC in RA patients,inhibiting or inducing PBMC ferroptosis in RA patients,will inhibit or promote the secretion of inflammatory factors.Inhibition of PBMC ferroptosis in RA patients may be helpful in the treatment of RA.
10.PI3K/AKT/mTOR in synovial fluid extends the proinflammatory response of macrophage polarization in knee osteoarthritis
Zong Jiang ; Tengxun Guo ; Weiya Lan ; Fang Tang ; Wukai Ma ; Jia Liu
Acta Universitatis Medicinalis Anhui 2024;59(3):377-383
Objective :
Given that the PI3K/AKT/mTOR signaling pathway is associated with the progression of knee osteoarthritis (KOA) , this study aims to investigate whether the polarization induction of synovial macrophages mediated by the PI3K/AKT/mTOR signaling axis is the cause of KOA progression .
Methods :
The synovial fluid of KOA KL-Ⅱ and KL-Ⅲ patients and normal individuals was collected , and the percentage of M1 macrophages (CD80 , CD86) and M2 macrophages (CD163 , CD206) in the synovial fluid (M1 /M2 ratio) was measured to e- valuate the polarization of macrophage cytokines such as IL-1 , IL-6 , IL-10 , and tumor necrosis factor (TNF) -α, transforming growth factor ( TGF)-βExpression in KOA synovial fluid , and detect and analyze of key molecules PI3K/AKT/mTOR signaling axis PI3K , AKT3 , mTORC1 , and inducible nitric oxide synthase ( iONS) in KOA synovial fluid .
Results :
Compared with the synovial fluid of normal individuals , the percentage of M1 macrophages (CD80 , CD86) in KOA patients increased (P < 0. 01) , and the M1 /M2 ratio increased ( P < 0. 001) ; The ex- pression of IL-1 , IL-6 , and TNF-αin the synovial fluid of the KOA group was also higher than that of the control group (P < 0. 01) , while the expression of IL-10 and TGF-βin the KOA group was significantly reduced ( P < 0. 01) ; The key proteins PI3K , AKT3 , mTORC1 , and downstream inflammatory factor iONS in the PI3K/AKT/ mTOR signaling pathway in the synovial fluid of the KOA group were higher than those in the control group (P < 0. 01) .
Conclusion
In KOA synovial fluid , M1 macrophage polarization plays a dominant role , and the inflam-
matory response mediated by M1 macrophage polarization may be the cause of synovitis . At the same time , the PI3K/AKT/mTOR signaling pathway may mediate the polarization of M1 macrophages involved in KOA inflammato- ry response .