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Abstract Introduction The relationship between humidity and systemic lupus erythematosus (SLE) has yielded inconsistent results in prior research, while the effects of humidity on lupus in animal experiments and its underlying mechanism remain inadequately explored. Methods The present study aimed to investigate the impact of high humidity (80 ± 5%) on lupus using female and male MRL/lpr mice, with a particular focus on elucidating the role of gut microbiota in this process. To this end, fecal microbiota transplantation (FMT) was employed to transfer the gut microbiota of MRL/lpr mice under high humidity to blank MRL/lpr mice under normal humidity (50 ± 5%), allowing for an assessment of the effect of FMT on lupus. Results The study revealed that high humidity exacerbated lupus indices (serum anti-dsDNA, ANA, IL-6, and IFN- g, and renal pathology) in female MRL/lpr mice but had no significant effect on male MRL/lpr mice. The aggravation of lupus caused by high humidity may be attributed to the increased abundances of the Rikenella, Romboutsia, Turicibacter, and Escherichia-Shigella genera in female MRL/lpr mice. Furthermore, FMT also exacerbated lupus in female MRL/lpr mice but not in male MRL/lpr mice. Conclusion In summary, this study has demonstrated that high humidity exacerbated lupus by modulating gut microbiota in female MRL/lpr mice. The findings underscore the importance of considering environmental factors and gut microbiota in the development and progression of lupus, particularly among female patients.
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Objective:To investigate the effects of rapamycin (RAPA) on the cognitive function of lupus mice by regulating Cysteine rich 61 (Cyr61) and autophagy.Methods:MRL/lpr lupus mice were randomly divided into lupus group and rapamycin + lupus group, wild-type C57BL/6 mice were randomly divided into normal control group and rapamycin group with six mice in each group, RAPA + lupus group and rapamycin group were intraperitoneally injected with RAPA (2.0 mg/kg). The lupus group and the normal control group were injected with equal amounts of dimethyl sulfoxide (DMSO). Morris water maze was used to observe the cognitive function of mice. Western blotting was used to detect the expression of Cyr61, Programmed cell death-1 (Beclin-1), Microtubule associated protein 1 light chain3B (LC3B). Hematoxylin-eosin (HE) staining was used to observe the pathological changes in the hippocampus. The changes of neurons and bodies in hippocampus were observed by Nissl staining. The localization and expression of Cyr61 and LC3B in hippocampus were detected by immunofluorescence staining. One-way analysis of variance (ANOVA) was used between groups, and LSD-T test was used for pairwise comparison.Results:Western blotting results showed thatthe protein expression of Cyr61 (0.99±0.15) was significantly increased ( P=0.011), and the protein expression of Beclin-1 (0.64±0.04) and LC3B(0.54±0.05) was significantly decreased in lupus group ( P=0.025, P= 0.008) when compared with normal control group (0.73±0.08, 0.81±0.12, 0.80±0.03). The expressions of Cyr61 (0.75±0.05, 0.75±0.08), Beclin-1 (0.84±0.08, 0.92±0.04) and LC3B (0.93±0.16, 0.76±0.08) in rapamycin group and rapamycin + lupus group were not significantly changed ( P>0.05). Compared with rapamycin group, the protein expression of Cyr61 (0.99±0.15) was significantly increased ( P=0.016), and Beclin-1 (0.64±0.04), LC3B (0.54±0.05) was significantly decreased in lupus group ( P=0.013, P=0.001). The expressions of Cyr61 (0.75± 0.08), Beclin-1 (0.92±0.04) and LC3B (0.76±0.08) were not significantly changed in rapamycin+lupus group ( P=0.999, P=0.241, P=0.062). Compared with lupus group, the expression of Cyr61 (0.75±0.08) protein in rapamycin+lupus group was significantly decreased ( P=0.016), and the expression of Beclin-1 (0.92±0.04) and LC3B(0.76±0.08) protein were significantly increased ( P=0.002, P=0.017). Immunofluorescence results showed that Cyr61 and LC3B were mainly expressed in the cytoplasm of hippocampal neurons, and the quantitative detection results were consistent with western blot results, the differences were statistically significant ( P=0.025, P=0.032). HE staining showed that the levels and number of cells in the hippocampus of mice with lupus were reduced, and the arrangement was sparse, and the nuclei were hyperchromatic, showing nuclear pyknosis and migration. The results of Nissl staining showed that there were relatively fewer Nissl bodies, loose arrangement of neurons and vacuolar areas in some cells, which were improved after RAPA treatment in lupus mice. Conclusion:RAPA can protect the cognitive function of lupus mice by inhibiting the expression of Cyr61 in hippocampus and promoting autophagy.
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Aim: To explore the role of paeoniflorin-6'- O-benzenesulfonate (CP-25) in MRL/lpr mice and the regulating action on Thl7 cell differentiation. Methods MRL/lpr mice were randomly assigned to five groups as follows; model group, CP-25 (20, 40, 80 mg · kg
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Objective This study aims to investigate the variation of neutrophil extracellular traps (NETs) in animal model of staphylococcus aureus (SA) infected systemic lupus erythematosus (SLE) MRL/Lpr mouse.Methods The infection model was developed by injecting the Staphylococcus Aureus in the mouse tail vein.The Staphylococcus Aureus load of the serum and kidney was detected by enzyme-linked immuno sorbent assay (ELISA).The level of NETs complex in the serum was detected by Cell-Death-Elisa-Plus-Kit and anti-myeloperoxidase (MPO) antibody.The kidney NETs formation was tested with the immunofluorescence.Statistical program for social sciences (SPSS) 17.0 software and Image Pro Plus statistical software were used for analysis.Comparisons between groups were made using unpaired t test for normally distributed numerical data,nonparametric test for non-normally distributed numerical data,and single factor analysis (ANOVA) for variance.Results The incidence of walking instability,purulent exudate in eyes and death was 2/6,1/6,1/6 respectively in Staphylococcus Aureus-infected MRL/Lpr mice,However,The control group had no death and related symptoms.The Staphylococcus Aureus load of the serum was higher in MRl/lpr mice when compared to the control group [(106.79±23.39) ng/ml vs (48.82±11.49) ng/ml,U=2.739,P＜0.05] after Staphylococcus Aureus infection.The control group had a higher NETs level [(2.24±0.15) vs (1.64±0.08),U=2.882,P＜0.05],however,all the MRL/Lpr mice had no significant change of the NETs level [(2.63±0.61) vs (2.65 ±0.260),U=0.548,P＞0.05] after Staphylococcus Aureus infection.Conclusion After infected by the Staphylococcus Aureus,more symptoms,higher death rate and higher Staphylococcus Aureus load of the serum present in the MRlL/lpr mice than the control group.While,there's none significant change of the NETs level in Staphylococcus Aureus-infected MRLL/lpr mice.This suggests that the dyspoiesis of NETs is present in Staphylococcus Aureus-infected MRL/lpr mice.
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Objective To explore the immune mechanism in the systemic lupus erythematosus MRL/lpr mice at different months of age, and to provide the basis for research of its pathogenesis. Methods 3-,4-,5- and 6-month old female MRL/lpr mice, and wild type C57 female mice were used in this study, 10 mice per each group. Their organ coefficients were determined. ELISA was performed to detect the serum levels of double stranded DNA(ds-DNA) antibody. The interleukins IL-2,IL-4,IL-17 and tumor necrosis factor-α(TNF-α)in spleen tissue were detected. Flow cytometry was used to assess the content of spleen lymphocyte CD3 cells and CD4/CD8 cell ratio. Results Compared with the 3-month old wild-type C57 mice,the spleen coefficient,the blood concentration of ds-DNA antibody,IL-2 and TNF-α in the 3- to 6-month old MRL/lpr mice were significantly increased(P < 0.05). There was no significant difference between the concentrations of interleukin IL-4(P> 0.05). The blood concentration of IL-17 in the 5- and 6-month old MRL/lpr mice was significantly lower(P < 0.05 or P < 0.01)than that in the 3-month old MRL/lpr mice. The rest indexes of MRL/lpr mice showed no obvious changes or significant difference in the mice at different ages. Compared with the 3-month old C57 mice,the spleen CD3 lymphocyte concentration in the MRL/lpr mice was significantly decreased(P< 0.01). With the increasing age, the CD3 lymphocyte concentration and D4 +/CD8 +cell ratio in the MRL/lpr mice were decreased, however, showing a non-significant difference(P ﹥0.05). Conclusions The data obtained in this study indicate that 3-month old lupus MRL/lpr mice have already immune injury,increasing with the increase of age of the mice.
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Objective:Preliminary study on therapeutic effects of adipose tissue derived stem cells (ADSCs) on MRL/lpr mice and the effect on imbalance of Th17/Treg.Methods:Fifteen 12-week-old MRL/lpr mice were randomly divided into 3 groups by using random number table,including ADSCs group,control group and cyclophosphamide (CTX) group,with 5 in each group.ADSCs group and control group were injected with 1 × 106ADSCs or phosphate buffered solution (PBS) via tail vein respectively,once a week,a total of eight times.CTX group was injected CTX at a dose of 15 mg/kg body weight,once a week for 2 weeks,and then repeated after 2 weeks' rest,a total of four times.The 24-hour proteinuria was measured before and after treatment.All the mice were sacrificed after treatment for 8 weeks.Th17 cells and Treg cells in splenic were examined by flow cytometry.Results:(1) The 24-hour proteinuria in the three groups had no significant difference before treatment (P ＞ 0.05).After therapy for 4 weeks,the 24-hour proteinuria in the ADSCs and CTX groups was much lower than those in control group,and the difference was significant [(5.02 ± 1.61) g/L vs.(7.10 ± 1.63) g/L,(4.90 ±0.71) g/L vs.(7.10 ± 1.63) g/L,P ＜ 0.05],and the longer the duration of treatment (8 weeks),the more obvious effect [(2.24 ± 0.73) g/L vs.(10.36 ± 1.64) g/L,(3.80 ± 1.45) g/L vs.(10.36 ± 1.64) g/L,P ＜0.01].There was no significant difference in 24-hour proteinuria between ADSCs group and CTX group (P ＞ 0.05).(2) Percentage of Treg cells/CD4 + T cells in the spleen lymphocytes:The percentages in ADSCs and CTX groups were higher than that in control group.The levels were 13.62％ ± 1.87％,14.14％ ± 1.29％,10.71％ ± 1.23％,respectively,but there was no significant difference (P ＞ 0.05).(3) Percentage of Th17 cells/CD4 +T cells in the spleen lymphocytes:The percentages in ADSCs and CTX groups were significantly lower than that in control group.The levels were 1.43％ ± 0.20％,1.63％ ± 0.65％,6.37％ ± 1.64％,respectively,with statistical significance (P ＜ 0.01).Conclusion:Transplantation of ADSCs can reduce the 24-hour proteinuria in MRL/lpr mice.To prolong the time of treatment,the effect is more significant.Transplantation of ADSCs can up-regulate Treg cells and down-regulate Th17 cells.ADSCs have the ability to regulate the immune balance of Th17/Treg in MRL/lpr mice,suggesting that ADSCs play the role of anti-inflammatory and immune regulation by regulating the Treg and Th17 cells.
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AIM:To investigate the effect of arsenic trioxide (ATO) on T-bet/GATA3 signal pathway in MRL/lpr mice.METHODS:MRL/lpr mice and C57BL/6J mice at the age of 20 weeks were chosen and then divided in 2 different sub-groups,respectively.The mice in 2 sub-groups received ATO (0.4 mg · kg-1.d-1) and sodium chloride (NS,volume weight-determined) by intraperitoneal injection respectively for 2 months.Afterward,the spleens were isolated fron the MRL/lpr and C57BL/6J mice under pathogen-free condition and the suspensions were prepared.The mRNA level of T-bet,GATA3,IFN-γ,IL-4 and the mRNA ratio of T-bet/GATA3 were detected by RT-qPCR.The protein expression of T-bet and GATA3 was determined by Western blot.The serum levels of IFN-γ and IL-4 were measured by ELISA.RESULTS:The mRNA and protein levels of T-bet,IFN-γand the mRNA ratio of T-bet/GATA3 in NS group of MRL/lpr mice were higher than those in NS group of C57BL/6J mice (P ＜0.05).However,the GATA3 and IL-4 were lower in NS group of MRL/lpr mice in both mRNA and protein level (P ＜ 0.05).In MRL/lpr mice,the mRNA and protein levels of T-bet,IFN-γ and the mRNA ratio of T-bet/GATA3 were lower in ATO group compared with NS group (P ＜ 0.05),no difference was found in GATA3 and IL-4.No difference of the indexes mentioned above between ATO group and NS group in C57BL/6J mice was observed.CONCLUSION:ATO may affect the signaling pathway of T-bet/GATA3 to down-regulate the mRNA expression and the protein secretion of IFN-γ by decreasing the expression of T-bet in MRL/lpr mice.
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AIM:To investigate the effect of arsenic trioxide (ATO) on T-bet/GATA3 signal pathway in MRL/lpr mice.METHODS:MRL/lpr mice and C57BL/6J mice at the age of 20 weeks were chosen and then divided in 2 different sub-groups,respectively.The mice in 2 sub-groups received ATO (0.4 mg · kg-1.d-1) and sodium chloride (NS,volume weight-determined) by intraperitoneal injection respectively for 2 months.Afterward,the spleens were isolated fron the MRL/lpr and C57BL/6J mice under pathogen-free condition and the suspensions were prepared.The mRNA level of T-bet,GATA3,IFN-γ,IL-4 and the mRNA ratio of T-bet/GATA3 were detected by RT-qPCR.The protein expression of T-bet and GATA3 was determined by Western blot.The serum levels of IFN-γ and IL-4 were measured by ELISA.RESULTS:The mRNA and protein levels of T-bet,IFN-γand the mRNA ratio of T-bet/GATA3 in NS group of MRL/lpr mice were higher than those in NS group of C57BL/6J mice (P ＜0.05).However,the GATA3 and IL-4 were lower in NS group of MRL/lpr mice in both mRNA and protein level (P ＜ 0.05).In MRL/lpr mice,the mRNA and protein levels of T-bet,IFN-γ and the mRNA ratio of T-bet/GATA3 were lower in ATO group compared with NS group (P ＜ 0.05),no difference was found in GATA3 and IL-4.No difference of the indexes mentioned above between ATO group and NS group in C57BL/6J mice was observed.CONCLUSION:ATO may affect the signaling pathway of T-bet/GATA3 to down-regulate the mRNA expression and the protein secretion of IFN-γ by decreasing the expression of T-bet in MRL/lpr mice.
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[Objective] To observe the effect of Jiedu Quyu Ziyin decoction on TLR4 signaling pathway in macrophages of MRL/lpr lupus mice. [Method] MRL/lpr lupus mice were divided into four groups:model group, prednisone group, Jiedu Quyu Ziyin decoction group(hereinafter referred to as:Chinese medicine group) and prednisone plus Chinese medicine group (hereinafter referred to as:combination of Chinese and western medicine group). The mice were gavaged with saline, prednisone, Jiedu Quyu Ziyin decoction and prednisone added Jiedu Quyu Ziyin decoction for 4 weeks. Macrophages of lung, peritoneal and spleen were collected and the expression of related genes was detected by RT-PCR. [Result] TLR4 mRNA in lung macrophages, and TLR4 protein in splenic macrophages increased significantly(P<0.05) after the treatment of prednisone. The increased TLR4 protein in splenic macrophages was significantly decreased by combining with Jiedu Quyu Ziyin decoction(P<0.05). Prednisone can significantly reduce the TLR4 downstream molecules such as MyD88, IFN-α, iNOS mRNA expression in lung and peritoneal macrophages(P<0.05). The decreased MyD88 mRNA in lung macrophages was increased significantly by combining with Jiedu Quyu Ziyin decoction(P<0.05). [Conclusion] TLR4 signaling pathway is changed in macrophages after glucocorticoid administration in MRL/lpr lupus mice. Jiedu Quyu Ziyin decoction can reduce the abnormal glucocorticoid-induced TLR4 protein expression.
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Objective:To observe the effect of CD40 siRNA on expression of IFN-γ,IL-17,IL-4 and anti-dsDNA antibody of systemic lupus erythematosus (SLE)animal model MRL/Lpr mice and to discuss its therapy on MRL/Lpr mice.Methods:In the study,16 female MRL/Lpr mice were randomly divided into control group (n =4),empty vector group (n =4),CD40-siRNA1 group (n =4)and CD40-siR-NA2 group (n =4).The vectors expressing siRNA against CD40 were injected by tail veil into MRL/Lpr mice,while MRL/Lpr mice in control group and empty vector group were injected with the same dose of PBS and pGFP-V-RS vector respectively.The injection was given six times and every one day.The mice were sacrificed 14 d after injection,and the spleen tissue was weighed.The pGFP-V-RS was labeled by green fluorescent protein(GFP)and the tissue sections were observed whether siRNA expressed in the spleen.The expression levels of IFN-γ,IL-17,IL-4 and anti-dsDNA antibody in the sera were detected by ELISA method on the 1st day before the first time and the 2nd,5th,8th,11th,and 14th days after last injection,and the expression levels of CD40 mRNA in spleen tissue of MRL/Lpr mice were detected by RT-PCR and the expression levels of CD40 protein in spleen tissue of MRL/Lpr mice were detected by immunohistochemistry method.Results:The expression vector of CD40-siRNA could express in the spleen of MRL/Lpr.The spleens in CD40-siRNA1 group [(78.85 ±5.61 )mg]and CD40-siRNA2 group [(80.25 ±4.07)mg]were lower than those in control [(141.88 ±7.81)mg]and empty vector group [(153.10 ±7.60)mg].The levels of IL-17,IFN-γand anti-dsDNA antibody were lower and the levels of IL-4 was higher in CD40-siRNA1 group and CD40-siRNA2 group on the 2nd,5th and 8th days after last injection than on the 1st day before the first time (P 0.05). Though the levels of anti-dsDNA antibody in CD40-siRNA1 group and CD40-siRNA2 group on the 11th day was higher than on the 8th day,there was more significance than those in control group and empty vector group (P <0.05).There was no significance between the 4 groups on the 14th day.The levels of CD40 mRNA and protein were lower in CD40-siRNA1 group and CD40-siRNA2 group than in control group and empty vector group on the 14th day after last injection (P <0.05).Conclusion:CD-40 si-RNA can reduce the concentration of IL-17,IFN-γand of anti-dsDNA antibody in serum,and at the same time,it can elevate the concentration of IL-4 and suppress CD40 mRNA and protein of spleen in MRL/Lpr.Meanwhile after suppressing CD40 mRNA and protein,it can reduce inflammatory response of the mice and the disease activity of MRL/Lpr,suggesting that CD-40 siRNA has therapy effect on SLE.
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BACKGROUND:The non-specific immune suppression method is generaly used for treatment of systemic lupus erythematosus, but poor prognosis, such as infection and high recurrence rate, exists. OBJECTIVE:To evaluate the therapeutic effect of bone marrow mesenchymal stem cel transplantation on systemic lupus erythematosus in mice. METHODS:Sixteen mice with systemic lupus erythematosus were equivalently randomized into control and experimental groups, or then subjected to passage 3 bone marrow mesenchymal stem cel transplantation or the equal volume of normal saline via the tail vein, respectively. Mouse urine samples were colected to detect urine protein levels by Bradford method. Blood samples from the tip of the mouse tail were extracted to detect serum anti-ds-DNS antibody concentration by radioimmunoassay. Mouse kidney tissues were taken and observed pathohistologicaly through hematoxylin-eosin staining and immunohistochemistry staining under microscope. Flow cytometry was used to detect the expression of CD4+CD25+T cels in the inner canthus blood, fresh spleen and thymus. RESULTS AND CONCLUSION:Within 10 weeks after cel transplantation, the urine protein levels in the two groups were gradualy increased, and the rising velocity was higher in the control group than in the experimental group. From the 4th to 10th week, the urine protein levels in the experimental group were significantly lower than those in the control group (P 0.05). The serum anti-ds-DNA antibody concentration in the experimental group was significantly lower than that in the control group (P < 0.05). Taken together, bone marrow mesenchymal stem cel transplantation can improve the pathological damage in systemic lupus erythematosus mice, and has a certain therapeutic effect on systemic lupus erythematosus.
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Objective:To explore the change of B cell numbers in active MRL/lpr lupus mice , and their regulation mechanisms.Methods:B cell cycle and the percent of B cells in spleen lymphocytes of active MRL /lpr lupus mice and normal C 57/B6 mice were analyzed by using flow cytometry .The apoptotic B cells and their subclass were analyzed by Annexin V and PI staining.Further more ,B cells were purified by magnetic sorting , and real-time quantitative PCR was carried out to detect apoptosis-related gene.Results:Compared with the C57/B6 mice,the percent of B cells in active MRL/lpr lupus mice were significantly reduced (P<0.01),while the percent of apoptotic cells were significantly increased (P<0.01).The percent of early apoptotic B cells were sig-nificantly increased ( P <0.01 ) which including the immature and mature B cells , while the late apoptotic B cells were unchanged.Further more,we found that the anti-apoptotic protein BIRC3 was significantly reduced in active lupus B cells (P<0.01), while the pro-apoptotic protein BCL2L1 and BBC3(PUMA) were significantly increased(P<0.01).Conclusion: B cells in active lupus mice were significantly reduced while early apoptotic B cells were increased , which may be attributed to the changed balance between the anti-apoptotic and pro-apoptotic proteins , suggesting the reduction of B cells in SLE patients may be related to their increased early apoptosis .
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Objective To investigate the effects of artesunate (ART) on interstitial pneumonia and sialadenitis in MRL/lpr mice.Methods A total of 18 MRL/lpr mice were randomly allocated to a hydroxychloroquine sulfate (HCQ) group,a ART group and a control group.At the age of 18 weeks,the mice in the HCQ group and ART group were given HCQ 150 mg/kg daily and ART 50 mg/kg daily for 12 weeks,respectively.The histopathological changes of pneumonitis and submaxillaritis were assessed by hematoxylin and eosin staining.The levels of monocyte chemoattractant protein-1 (MCP-1) in the serum and urine were measured by the enzyme-linked immunosorbent assay.Results At the age of 30 weeks,the index of peribronchiolar lesion (1.62 ± 0.19,1.52 ± 0.30 vs.1.95 ± 0.34;all P＜0.05),the index of perivascular lesion (1.23 ± 0.18,1.28 ± 0.12 vs.1.57 ± 0.33;all P＜0.05),the alveolar lesions index (1.35 ± 0.16,1.05 ± 0.15 vs.1.72 ± 0.34;all P＜0.05) and the submaxillaritis index (1.48 ± 0.22,1.43 ± 0.15 vs.1.84 ± 0.34;all P＜0.05) in the HCQ group and the ART group were significantly decreased than those in the control group.The MCP-1 levels in the serum (1 103.02 ± 185.56 pg/ml,1 072.37 ± 242.43 pg/ml vs.1 490.67 ± 329.43 pg/ml;all P＜0.05) and urine (189.16 ± 70.85 pg/ml,198.79 ± 113.47 pg/ml vs.446.79 ± 192.31 pg/ml;all P＜0.05) in the HCQ group and the ART group were significantly lower than those in the control group.Conclusion ART can decrease the MCP-1 level,and ameliorate interstitial pneumonitis and sialadenitis in MRL/lpr mice.
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Objective:To inspect the relationship between the therapeutic effect of DHA on lupus nephritis and the negative immune regulation of TLR4/NF-κB signal pathway which was induced by SIGIRR;in vitro,to observe the effect of DHA on damaged HK-2 cell.Methods: In vivo,MRL/lpr mice were divided in model group,DHA groups(25,50,100 mg/kg),positive group (prednisone,5 mg/kg),and C57BL/6 mice were taken as control group.Administrate drugs daily for 12 weeks.Examine the changes in renal pathology;the expression of SIGIRR,IRAK1,TRAF6 in kidneys were determined by Western blot.In vitro,treat human renal tubular epithelial cell HK-2 cells with LPS ,and co-culture cells with DHA at the concentration of 0.67 μg/ml to 6.00 μg/ml for 6 h, 12 h and 24 h.Detect SIGIRR expression by Western blot and the level of IL-6 and CCL2 of HK-2 cells by ELISA.Results:In vivo, renal pathology revealed that kidneys of model group were damaged , while treatment with 100 mg/kg DHA alleviated renal injury.Compared to model group ,SIGIRR expression of DHA 100 mg/kg group increased a little ,and the expression of this protein had a tendency to increase with the augment of DHA dose .In vitro,DHA treatment reduced secretion of CCL 2 in HK-2 cells,and treatment of 0.67 μg/ml DHA for 24 h increased SIGIRR expression significantly , which also showed a growing expression with time.Conclusion:DHA could inhibit development of mouse lupus nephritis through increasing SIGIRR expression which inhibited TLR4/NF-κB signal pathway;DHA inhibited CCL2 secretion of HK-2 cells which were irritated by LPS ,and it may be associated with increased expression of SIGIRR .
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ObjectiveToclarifytheprotectiveactionofaChinesemedicine,Jie-Du-Qu-Yu-Zi-Shen prescription, on the lupus nephritis in MRL/lpr mice through the TLR9-MyD88-NF-κB signaling pathway .Methods Thirty MRL/lpr mice were randomly divided into model group , prednisone group , and traditional Chinese medicine ( TCM) group (n=10).Each treatment group received appropriate treatment or drug therapy for 8 weeks.The pathological changes of mouse renal tissues in the three groups were examined and compared using HE , PAS, and Masson staining .Real-time PCR was performed to compare the expression of TLR-9, MyD88, NF-κBm RNA mRNA in each group of the mice . Results Compared with the model group , the Chinese medicine Jie-Du-Qu-Yu-Zi-Shen prescription significantly improved the renal pathological tissue damages in the MRL/lpr mice.Real-time PCR assay showed that the expression levels of TLR-9, MyD88, NF-κB mRNA in the TCM group were significantly lowered (P0.05 ) .Conclusions Jie-Du-Qu-Yu-Zi-Shen prescription has protective effect on the kidney of MRL /lpr mice, alleviating its pathological changes , and delaying the disease course, probably, through inhibiting the TLR-9-MyD88-NF-κB signaling pathway in the kidneys of MRL/lpr mice.
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Objective:To observe the effect of CD40 siRNA on the changes in kidney,urinary protein and complement C3 of MRL/Lpr mice and explore its therapy on lupus nephritis.Methods: 16 female MRL/Lpr mice were randomly divided into control group,empty vector group,CD40-siRNA1 group and CD40-siRNA2 group.The vectors expressing siRNA against CD40 were injected by tail veil into MRL/Lpr mice,while MRL/Lpr mice in control group and empty vector group were injected with the same dose of PBS and pGFP-V-RS vector respectively.There were six times injection and every one day.The 24 hours urine output was gathered 24 hours before mice were killed.Fourteen days following administration,these mice were killed,tissue sections of kidney were observed if the signal of siRNA were expressed in kidney.The expression levels of CD40 mRNA and protein in kidney tissue of MRL/Lpr mice were detected by RT-PCR and immunohistochemistry methods respectively.At the same time,the pathological changes of the kidney were observed by haematoxylin-eosin ( HE) staining method.The 24 h urinary protein content was detected using the method of coomassie brilliant blue and the expression levels of complement C3 in serum were detected by Immunoturbidimetric assays.Results:The vector of CD40-siRNA was expressed in kidney of MRL/Lpr.The expression levels of CD40 mRNA and protein in kidney were lower in CD40-siRNA1 group and CD40-siRNA2 group than control group and empty vector group on the 14th day after last injection ( P<0.05).The inflammatory cells infiltration of kidney and some glomerular volume were significantly reduced in CD40-siRNA1 group and CD40-siRNA2 group than control group and empty vector group.The renal tubular swelling was alleviated in CD40-siRNA1 group and CD40-siRNA2 group than control group and empty vector group.The levels of 24 hours urinary protein were lower and the levels of complement C3 were higher in CD40-siRNA1 group and CD40-siRNA2 group than control group and empty vector group ( P<0.05). Conclusion:CD-40 siRNA can suppress the expression levels of CD40 mRNA and protein and decrease inflammatory cells infiltration in kidney of MRL/Lpr.Meanwhile after suppressing expression of CD40 mRNA and protein, it can reduce the content of 24 hours urinary protein and elevate the level of complement C3 in serum,which CD-40 siRNA can delay progress of the disease and protect kidney,so that it has therapy effect on lupus nephritis.
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Objective To investigate the efficacy and adverse reactions of half-dose glucocorticosteroid and cytoxan,combined with leflunomide for the treatment of lupus nephritis (LN) of MRL/lpr mice,and provide experimental evidences for LN therapy.Methods Twenty-eight 10-week-old MRL/lpr mice were randomly divided into four groups:Group A,blank control group; Group B,classical control group; Group C,full-dose control group; Group D,half-dose treatment group,with 7 mice in each group.The therapeutic efficacy and side reactions in the four groups were observed and compared before and 12 weeks after treatment.Statistical analysis was conducted with one-way ANOVA,q test and Pearson's correlation analysis.Results The serum anti-double stranded DNA (anti-dsDNA) antibody titers (0.43±0.16,0.32±0.09,0.44± 0.18,1.95±0.19) U/ml,serum creatinine level (1.63±0.63,0.40±0.23,0.82±0.21,10.86±2.17) mg,24-hour urine protein excretion level (71±8,60±5,68±3,121±10) μmol/L and renal pathological changes in group B,C,D were significantly improved than those of the group A (P＜0.05) after 12 weeks treatment.There was no significant difference in the efficacy between group B,C,and D (P＞0.05).The incidence of adverse reactions in group D was significantly lower than that in other groups (P＜0.05).Conclusion Multi-target therapy,such as half-dose prednisone and CTX,combined with leflunomide can effectively control lupus disease activity with less side effects.This regimen is cheap,safe and effective for the treatment of LN in MRUL/lpr mice.This study has provided animal evidences for this multi-target therapy for LN.
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Objective To detect the expression of Blimp1 gene in MRL/lpr mice,so as to provide new ideas for plasma cell-targeting therapy of systemic lupus erythematosus (SLE).Methods Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation analysis (ChIP) were performed to evaluate the regulatory effect of Blimp1 on B cell maturation antigen (BCMA),and fluorescence-based quantitative PCR was carried out to detect the expression of Blimp1 mRNA in spleen and lymph node tissue of MRL/lpr mice and normal control mice.The intergroup difference in Blimp l mRNA expression was assessed by rank sum test.Results Blimp1 could bind to the BCMA gene promoter.Increased Blimp1 mRNA expression was observed in both the spleen and lymph node tissue of MRL/lpr mice compared with the normal control mice (Z =2.609,3.402,respectively,both P ＜ 0.01).Conclusions BCMA appears to be the target gene of Blimp1,and Blimp1 may plays a certain role in the maintenance of plasma cell survival and antibody production via directly regulating BCMA gene expression.
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Objective To explore the effects of Jiedu Quyu Ziyin Recipe (JQZR) on the apoptosis and expressions of bcl-2 and bax mRNA of peripheral-blood lymphocyte in MRL/lpr mice. Methods 80 MRL/lpr mice were randomly divided into model group,TCM group, Western medicine group and TCM and Western medicine group,20 mice in each group, meanwhile,20 Kunming mice were selected as normal group, then intragastrically administered normal sodium, JQZR apozem, prednisone suspension and JQZR apozem and prednisone suspension, 0. 5ml every time,once daily for 12 weeks respectively. At the end of the 12th week, peripheral-blood lymphocytes of every mice purified by gradient centrifugation were cultivated for 48 hours ,then the apoptosis was detected by flow cytometry. Furthermore,the expressions of bcl-2 and bax mRNA of peripheral-blood lymphocyte were detected by RT-PCR. Results At Oh or 48h,the apoptosis ratios of PBLC in normal group, TCM group,Western medicine group and TCM and Western medicine group are higher than model group and the differences are significant(P <0. 05 ,P <0. 01 ) ,while,at Oh,which are not significant within Western medicine group, normal group and TCM and Western medicine group ( P > 0. 05) ,even if which are significant between TCM group and TCM and Western medicine group or between model group and TCM and Western medicine group( P < 0. 05 ). At 48h, the differences of apoptosis ratios of PBLC are significant between Western medicine group and TCM and Western medicine group( P < 0. 05 ), and between TCM group and TCM and Western medicine group or between model group and TCM and Western medicine group which are significant, too( P < 0. 01 ). The ratios of bcl-2/bax mRNA of normal group,TCM group and TCM and Western medicine group are significantly lower than which of model group or Western medicine group ( P < 0. 05, P < 0. 05, P <0. 01 ,respectively) ,but there is no significant difference between which of model group and Western medicine group.Conclusion JQZR has adjustable effects on the apoptosis and expressions of bcl-2 and bax mRNA of peripheralblood lymphocyte in MRL/lpr Mice.
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Objective To investigate the efficacy of umbilical cord mesenchymal stem cells (UC-MSCs) transplantation in the treatment of the MRL/lpr mice. Methods Twenty four 18-week-old MRL/lpr female mice were divided into 3 groups:group 1 (G1) were transplanted with 1×106 UC- MSCs through caudal vein, group 2 (G2) were transplanted with 1×106 UC- MSCs three times and group 3 (G3) were treated with 0.5 ml normal saline as controls. Enzyme linked immunosorbent assay (ELISA) was used to measure the levels of serum anti-dsDNA antibodies. Twenty-four hours proteinuria and body weight were assessed every two weeks. The histopathology changes of the kidneys and lungs were observed. Results ① At the 25th weeks, the 24 hours proteinuria in group G1 (2.3±1.9) mg and G2 (1.8±1.4) mg was decreased than that in the control group (3.8±2.1) mg (P<0.05), and at the 27th weeks, that of groups G1 (2.5±1.5) mg and G2 (1.9±1.2) mg was also significantly decreased than in the control group (5.4±2.4) mg (P<0.01); ② From the 24th week, the body weight of groups G1 and G2 increased significantly than that of the control group (P< 0.05). At week 29, serum creatinine decreased significantly in both groups G1 (7.2±3.2) μmol/L and G2 (6.2±2.8) μmol/L than in the control group (12.5±2.3 ) μmol/L (P<0.05); ③One week after transplantation, the levels of anti-dsDNA antibodies in group G1 (46±11)×102 U/ml and G2(49×43)×102 U/ml were bothsignificantly decreased than those of the control groups (99±42)×102 U/ml (P<0.05) and the difference between group G2 (36±15)×102 U/ml and the controls (68±32)×102 U/ml was statistically significant; ④The nephron crescent formation in group G1 (0.12±0.07) and G2 (0.08±0.02) was significantly lower that of the control group (0.20±0.06) (P<0.05) and that of group G2 was significantly less that of froup G1 (P<0.05); ⑤ The interstitial pneumonitis was singnificantly milder in group G1 than group G2. Conclusions UC- MSCs is very effective in treating MRL/lpr mice. It is safe and free of rejection reactions.