Increment of N-cadherin Expression in Bone Marrow Mesenchymal Stem Cells Accompanies by Imparment of Bone Marrow B Lymphocyte Production in MRL/lpr Lupus Mice / 中国实验血液学杂志

OBJECTIVE@#To investigate the hematological changes in MRL/lpr lupus mice and detect N-cadherin expression in their bone marrow mesenchymal stem cell (BMMSC).@*METHODS@#Peripheral blood cell count was analyzed. The ratio of each lineage in bone marrow was analyzed by flow cytometry. Bone marrow CFU-pre-B, BFU-E and CFU-GM were detected by colony formation assay. Expression of N-cadherin in BMMSC was analyzed by Western blot before and after treatment with the BMP/Smad pathway aganist BMP-2 and inhibitor Noggin.@*RESULTS@#Hemoglobin, red blood cell count and hematocrit decreased in lupus mice, compared with C57BL/6 mice. The ratio of B220+ lymphocyte and the number of CFU-pre-B in bone marrow of lupus mice decreased. Expression of N-cadherin in BMMSC of lupus mice was higher than that in the control group. Expression of N-cadherin decreased in BMP-2-treated BMMSC and increased after Noggin treatment.@*CONCLUSION@#Hematological changes in lupus mice include anemia and impairment of bone marrow B cell production. The expression of N-cadherin in BMMSC of lupus mice increases which maybe involved in abnormal hemogenesis in MRI/Ipr lupus mice.

Expression of BATF, a member of the activator protein-1 family, in renal tissues of MRL/lpr mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of BATF, a member of the activator protein-1 family, in the renal tissues of mice with lupus nephritis.</p><p><b>METHODS</b>The renal tissues from 24-week-old female MRL/lpr mice and age- and sex-matched C57BL/6 mice were examined for BATF protein expressions using Western blotting and for expressions of BATF, RORγt and IL-17A mRNA using quantitative real-time PCR. The results of laboratory examinations were analyzed in relation with the histopathology of the mice.</p><p><b>RESULTS</b>The urinary protein and ds-DNA levels were significantly higher in MRL/lpr mice than in the control mice (P<0.05). Compared to normal control mice, MRL/lpr mice showed obvious glomerular fibrosis and inflammatory cell infiltrating with significantly increased BATF protein and mRNA expressions (P<0.05) and RORγt and IL-17 mRNA expressions in the renal tissues (P<0.05). In MRL/lpr mice, the expression of BATF mRNA was positively correlated with the RORγt and IL-17A mRNA expressions in the renal tissues.</p><p><b>CONCLUSION</b>The renal expressions of BATF protein and mRNA is increased in MRL/lpr mice. BATF may participate in the immunopathogenesis of lupus nephritis by enhancing Thl7 cell response.</p>

Regulation of UC-MSC on Immune Inflammatory Thrombophilia in MRL/lpr Mice / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the immune repair effect of umbilical cord mesenchymal stem cells (UC-MSC) on inflammatory disorders and thrombophilia state of MRL/lpr mice by detecting the expression change of peripheral blood CD4(+) CD25(+) T cells and the levels of plasma inflammatory cytokines TNF-α, IL-6 and thrombosis indicators TF, FIB.</p><p><b>METHODS</b>Twenty five MRL/lpr mice were divided into control (C) group, UC-MSC one time treatment (UT1) group and UC-MSC three time treatments (UT3) group. UC-MSC cell suspension was injecled via tail vein and these mice were feeded in SPF environment. The blood samples were taken from the mice every 2 weeks after 16(th) week. FCM was used to detect the expression of CD4(+) CD25(+) T cells in peripheral blood, ELISA assay was used to detect the levels of inflammatory cytokines TNF-α, IL-6 and thrombosis indicators TF, FIB.</p><p><b>RESULTS</b>The expression of peripheral blood CD4(+) CD25(+) T cells in treatment groups increased during 16(th) to 18(th) week, dropped and tended to be stable since 20(th) week, and lower than those in control group. The levels of plasma TNF-α and IL-6 in treatment group decreased since 16(th) week and significantly lower than those in control group (P < 0.05). The levels of plasma TF and FIB in treatment group decreased since 16(th) week. The level of plasma TF in treatment group was significantly lower than those in control group (P < 0.05) since 18(th) week.</p><p><b>CONCLUSION</b>UC-MSC can repair the immune inflammatory microenvironment disorders of MRL/lpr mice through its immunomodulatory effect. UC-MSC contribute to repair of immune inflammatory thrombophilia of MRL/lpr mice.</p>

Expression of STAT3 and PIAS3 in renal tissues of MRL/lpr mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of STAT3 and PIAS3 in the renal tissues of rats with lupus nephritis.</p><p><b>METHODS</b>The kidneys were harvested from 18-week-old female MRL/lpr mice (lupus nephritis model group) and age-matched female BALB/C mice (normal control group). The expressions of STAT3 and PIAS3 mRNA in the renal tissues were measured by real-time quantitative RT-PCR, and the protein levels of STAT3 and p-STAT3 were examined using Western blotting and immunohistochemistry. The laboratory indices and renal histopathology of the mice were also investigated.</p><p><b>RESULTS</b>The urinary protein, blood urea nitrogen (BUN) and creatinine levels were significantly higher in MRL/lpr mice than in the control mice (P<0.05). The renal histopathology of MRL/lpr mice was characterized by increased mesangiocapillary proliferation and prominent inflammatory infiltration in the tubulo-interstitium, which were absent in the kidneys of normal control mice. Compared with the control mice, MRL/lpr mice showed significantly increased STAT3 mRNA and p-STAT3 protein levels in the renal tissues (P<0.05) with significantly lowered levels of PIAS3 mRNA (P<0.01). No significant difference was noted in the total STAT3 protein levels between MRL/lpr and control mice.</p><p><b>CONCLUSION</b>MRL/lpr mice have significantly increased expressions of STAT3 mRNA and p-STAT3 protein and decreased expression of mRNA PIAS3 in the kidneys compared with BALB/C mice.</p>

Expression of high mobility group box chromosomal protein 1 in mice with lupus nephritis / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To determine the role of the novel proinflammatory cytokine high mobility group box chromosomal protein 1 (HMGB-1) in the pathogenesis of lupus nephritis.</p><p><b>METHODS</b>Serum levels of anti-dsDNA antibodies were determined by enzyme linked immunosorbent assay (ELISA). Renal morphologic features were examined by light microscopy, electron microscopy, and immunohistologic analyses. The mRNA expression of HMGB-1 and monocyte chemoattractant protein-1 (MCP-1) was detected by RT-PCR.</p><p><b>RESULT</b>MRL/lpr mice demonstrated characteristic alterations of serum immune parameters, with progressively increased anti-dsDNA antibodies with age, compared with age-matched C57BL/6J mice. MRL/lpr mice showed progressive development of renal damage, starting at 12 weeks of age and reached the peak at 20 weeks. The observed lesions included the presence of enlarged hypercellular glomeruli, with increased numbers of both resident cells and infiltrating leukocytes. Higher expression of HMGB-1 mRNA was found in MRL/lpr mice than what in C57BL/6J mice. Expression of HMGB-1 was positively correlated with that of MCP-1 mRNA.</p><p><b>CONCLUSION</b>The results demonstrate that the higher expression of HMGB-1 may contribute to the pathogenesis of lupus nephritis.</p>

Oxidation status and γ-glutamylcysteine synthetase expression in the kidney of MRL/lpr lupus mice / 南方医科大学学报

<p><b>OBJECTIVE</b>[corrected] To investigate the role of glutathione (GSH) and γ-glutamylcysteine synthetase (γ-GCS) in lupus nephritis.</p><p><b>METHODS</b>Spectrophotometry was used to measure the oxidative/anti-oxidative indices including malonyldialdehyed (MDA) and GSH in the kidney of MRL/lpr lupus mice. Quantitative PCR and Western blotting were used to detected the expression of γ-GCS.</p><p><b>RESULTS</b>The level of GSH was lowered whereas the level of MDA increased significantly in the kidney tissue of MRL/lpr lupus mice as compared with that in normal control mice. The expression of γ-GCS mRNA and protein was significantly decreased in MRL/lpr lupus mice (P<0.05).</p><p><b>CONCLUSION</b>MRL/lpr lupus mice have abnormal oxidative stress in the kidney tissue, where the expression of γ-GCS decreased to lead to reduced GSH production, damaged antioxidative capacity, and eventually exacerbation of oxidative damage in the kidney.</p>

Study on the regulation of Jiedu Quyu Zishen Recipe on glucocorticoid receptor alpha in the renal tissue of MRL/lpr mice / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the action mechanism of Jiedu Quyu Zishen Recipe (JQZR) on the signal transduction of glucocorticoid receptor alpha (GRalpha) in the renal tissue of MRL/lpr mice.</p><p><b>METHODS</b>Thirty MRL/lpr mice were randomly divided into three groups, i.e., the model group, the Western medicine group, and the Chinese medicine group, 10 in each. Besides, another 10 Kunming mice was taken as the normal control group. The pathological changes of the renal tissue were observed using HE staining. The expression of GRalpha was analyzed using Real-time PCR and Western blot. The effects of JQZR on the binding power of GRalpha to cyclophilin A were detected using co-immunoprecipitation.</p><p><b>RESULTS</b>The renal injury degree of MRL/lpr mice in the Western medicine group and the Chinese medicine group was alleviated. Compared with the model group, the relative quantitation of GRalpha mRNA and protein expressions in the renal tissue of mice in the Western medicine group decreased, while they increased in the Chinese medicine group, showing statistical difference (P < 0.01). JQZR could significantly elevate the binding potency of GRalpha to cyclophilin A.</p><p><b>CONCLUSION</b>Up-regulating the expression of GRalpha and enhancing mutual actions of GRalpha and cyclophilin A was one of JQZR's effects on improving the lesion of the renal tissue.</p>

Therapeutic effect of total glucosides of paeony on lupus nephritis in MRL/lpr mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the therapeutic effect of total glucosides of paeony (TGP) on lupus nephritis (LN) in MRL/lpr mice.</p><p><b>METHODS</b>MRL/lpr mice with lupus nephritis were randomized into model group and TGP group. The urinary protein content was detected using Coomassie brilliant blue, and the serum levels of IgG anti-double-stranded DNA (dsDNA) antibodies and antinuclear antibodies (ANA) were measured by enzyme-linked immunosorbent assay (ELISA). The changes in the renal pathology were examined microscopically, and the spleen and thymus were weighed to calculate the spleen and thymus indexes.</p><p><b>RESULTS</b>At 15 and 30 days after TGP administration, the urinary protein content in the TGP group was significantly lower than that in the model group (P<0.05). TGP treatment significantly lowered the serum levels of anti-dsDNA antibodies and ANA and the weight and index of spleen (P<0.05), resulting also in lessened renal pathology at 30 days after the administration. Compared to those before TGP treatment, the urinary protein content and the levels of anti-dsDNA antibodies and ANA decreased significantly at 15 and 30 days after TGP administration (P<0.05), while in the model group, the level of anti-dsDNA increased significantly without obvious changes in urinary protein content or ANA. At 30 days after TGP administration, the urinary protein content was significantly lowered in the TGP group as compared to that at 15 days (P<0.05), but the antibodies showed no significant changes.</p><p><b>CONCLUSION</b>TGP can reduce urinary protein content and serum levels of anti-dsDNA antibodies and ANA, and lessen renal pathology in MRL/lpr mice with lupus nephritis, suggesting its therapeutic effect on lupus nephritis.</p>

Effects of three Chinese herbal antidotes (Herba artemisiae annuae, Herba hedyotis diffusae and Rhizoma cimicifugae) and their different combinations on regulated on activation normal T cell expressed and secreated expression in MRL/lpr mice / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effects of three Chinese herbal antidotes, i.e. Herba Artemisiae annuae (A), Herba Hedyotis diffusae (H) and Rhizoma Cimicifugae (C), all were ingredients of Jiedu Quyu Ziyin Recipe, for adjusting the regulated on activation normal T cell expressed and secreated (RANTES), gene expression in serum and renal tissue of MRL/lpr mice.</p><p><b>METHODS</b>Fifty-four MRL/lpr mice were randomized into 9 groups, with 6 in each, and intragastrically infused with A, H, C, A+H, H+C, A+C, A+H+C (all in dosage-form of decoction), prednisone suspension and physiological saline, respectively for 12 weeks. RANTES expression in serum and renal tissue of animals were detected with ELISA and RT-PCR at the end of the study.</p><p><b>RESULTS</b>Levels of RANTES expression was significantly reduced in the prednisone treated group after treatment. Excepting no significant change being observed in the groups treated with A and C, the changes in the other groups were all milder than those in the group treated with A+H+C.</p><p><b>CONCLUSION</b>Chinese herbal antidotes A, H and C in combination can significantly inhibit the RANTES expression in serum and renal tissue of MRL/lpr mice.</p>

A critical evaluation of the effect of population size and phenotypic measurement on QTL detection and localization using a large F2 murine mapping population

Population size and phenotypic measurement are two key factors determining the detection power of quantitative trait loci (QTL) mapping. We evaluated how these two controllable factors quantitatively affect the detection of QTL and their localization using a large F2 murine mapping population and found that three main points emerged from this study. One finding was that the sensitivity of QTL detection significantly decreased as the population size decreased. The decrease in the percentage logarithm of the odd score (LOD score, which is a statistical measure of the likelihood of two loci being lied near each other on a chromosome) can be estimated using the formula 1 - n/N, where n is the smaller and N the larger population size. This empirical formula has several practical implications in QTL mapping. We also found that a population size of 300 seems to be a threshold for the detection of QTL and their localization, which challenges the small population sizes commonly-used in published studies, in excess of 60 percent of which cite population sizes <300. In addition, it seems that the precision of phenotypic measurement has a limited capacity to affect detection power, which means that quantitative traits that cannot be measured precisely can also be used in QTL mapping for the detection of major QTL.

Aging changes of gelatinase activity in kidney tissue of autoimmune MRL/lpr mice / 中华病理学杂志

<p><b>OBJECTIVE</b>To determine whether increased expressions of gelatinases occur with aging in vivo in kidney tissue of autoimmune MRL/lpr mice by in situ zymography.</p><p><b>METHODS</b>MRL/lpr mice at the age of 8 weeks, 16 weeks and 24 weeks were investigated. Kidney protein extracts were compared for activities of MMP-2/9 by gelatin zymography. Immunohistochemistry and SDS-PAGE gelatin zymography were used to determine the expressions and activities of gelatinase A (MMP-2) and gelatinase B (MMP-9). To determine the net gelatinase activities in murine lupus kidney, in situ zymography was used with autoradiographic emulsion as substrate.</p><p><b>RESULTS</b>Both gelatinase A and B were seldom detected in the kidney tissue in 8 week old mice, Increased expressions of both latent and activated form enzymes of MMP-2/9 were identified in kidney extraction by SDS-PAGE gelatin zymography and immunohistochemical staining showed both MMP-2 and MMP-9 were obviously up-regulated within glomerulus as well as tubular-interstitium in mice at the age of 16 and 24 weeks. In situ zymography showed markedly increased gelatinase activities in kidney tissue consistent with the results of immunohistochemical staining, it is mainly derived from MMPs and inhibited by EDTA but not by PMSF or aprotinin.</p><p><b>CONCLUSIONS</b>These in vivo results suggested that MMP-2/-9 expressions were significantly up-regulated with aging in murine lupus nephritis, which may play an important role in promoting the remodeling formation of ECM and thus contribute to the progression of renal damage in this model.</p>