Effective Evaluation of Lupus Nephritis in Rat Podocyte Cell Culture
碩士 === 國立中興大學 === 獸醫學系暨研究所 === 99 === Lupus nephritis (LN), an autoimmune disease, is the most common forms in systemic lupus erythematosus (SLE). The patients of LN are usually associated with proteinuria which is caused by podocyte’s foot processes effacement. In recent studies, the induction of u...
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ndltd-TW-099NCHU55410272018-04-10T17:21:06Z http://ndltd.ncl.edu.tw/handle/dsb99c Effective Evaluation of Lupus Nephritis in Rat Podocyte Cell Culture 大鼠腎臟足細胞探討狼瘡腎炎療效評估 Sih-Yong Zeng 曾思詠 碩士 國立中興大學 獸醫學系暨研究所 99 Lupus nephritis (LN), an autoimmune disease, is the most common forms in systemic lupus erythematosus (SLE). The patients of LN are usually associated with proteinuria which is caused by podocyte’s foot processes effacement. In recent studies, the induction of urokinase-type plasminogen activator receptor or CD78 (uPAR, CD78) signaling in podocyte leads to foot process effacement. The clinical studies show that Interleukin-6 (IL-6) has been implicated in LN. Blockade of IL-6 by interleukin-6 receptor monoclonal antibody (mAb IL-6R) will reduces proteinuria production in murine lupus. Nevertheless, there are no data concerning the effects of IL-6 and mAb IL-6R on the expression of uPAR of podocytes in LN. In this study, we firstly, cultured podocytes from isolated glomerulli in rat, and the podocyte will be identified by immunofluorescence staining of synaptopodin and podocin Ab. Podocytes (1×103/mL) were experimentally grouped into three groups, followed: normal control group, IL-6 (100 ng/mL) group and mAb IL-6R (25 μg/ml) combine with IL-6 (100 ng/mL) group, estimated their expression at different time point. Evaluation indicators of improved effect in LN will be:(1) analysis of uPAR-mRNA level by quantitative real time reverse transcription polymerase chain reaction (qRT-PCR), (2) counting podocytes migration number by transwell assay, (3) measurement of uPAR and phosphor-STAT3 (p-STAT3) protein expression by immunofluorescence staining. The results showed that IL-6 group up-regulated uPAR, p-STAT3 protein expression, podocyte migration and uPAR mRNA rather than normal control and mAb-IL-6R/IL-6 groups. mAb-IL-6R/IL-6 group down-regulated uPAR protein and uPAR mRNA rather than groups normal control and IL-6 groups. In conclusion that IL-6 was seen to significantly stimulate uPAR expression in vitro that suppose leading to podocyte foot process effacement and reduce proteinuria counterpart with in vivo. However, blocked IL-6 by mAb IL-6R will reduce uPAR expression and stable podocyte in glomerular basement may also decrease proteinuria. This study will provide more thraputic application in lupus nephritis and futurer animal and human trial research. Kwong-Chung Tung 董光中 2011 學位論文 ; thesis 46 zh-TW |
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碩士 === 國立中興大學 === 獸醫學系暨研究所 === 99 === Lupus nephritis (LN), an autoimmune disease, is the most common forms in systemic lupus erythematosus (SLE). The patients of LN are usually associated with proteinuria which is caused by podocyte’s foot processes effacement. In recent studies, the induction of urokinase-type plasminogen activator receptor or CD78 (uPAR, CD78) signaling in podocyte leads to foot process effacement. The clinical studies show that Interleukin-6 (IL-6) has been implicated in LN. Blockade of IL-6 by interleukin-6 receptor monoclonal antibody (mAb IL-6R) will reduces proteinuria production in murine lupus. Nevertheless, there are no data concerning the effects of IL-6 and mAb IL-6R on the expression of uPAR of podocytes in LN. In this study, we firstly, cultured podocytes from isolated glomerulli in rat, and the podocyte will be identified by immunofluorescence staining of synaptopodin and podocin Ab. Podocytes (1×103/mL) were experimentally grouped into three groups, followed: normal control group, IL-6 (100 ng/mL) group and mAb IL-6R (25 μg/ml) combine with IL-6 (100 ng/mL) group, estimated their expression at different time point. Evaluation indicators of improved effect in LN will be:(1) analysis of uPAR-mRNA level by quantitative real time reverse transcription polymerase chain reaction (qRT-PCR), (2) counting podocytes migration number by transwell assay, (3) measurement of uPAR and phosphor-STAT3 (p-STAT3) protein expression by immunofluorescence staining. The results showed that IL-6 group up-regulated uPAR, p-STAT3 protein expression, podocyte migration and uPAR mRNA rather than normal control and mAb-IL-6R/IL-6 groups. mAb-IL-6R/IL-6 group down-regulated uPAR protein and uPAR mRNA rather than groups normal control and IL-6 groups. In conclusion that IL-6 was seen to significantly stimulate uPAR expression in vitro that suppose leading to podocyte foot process effacement and reduce proteinuria counterpart with in vivo. However, blocked IL-6 by mAb IL-6R will reduce uPAR expression and stable podocyte in glomerular basement may also decrease proteinuria. This study will provide more thraputic application in lupus nephritis and futurer animal and human trial research.
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author2 |
Kwong-Chung Tung |
author_facet |
Kwong-Chung Tung Sih-Yong Zeng 曾思詠 |
author |
Sih-Yong Zeng 曾思詠 |
spellingShingle |
Sih-Yong Zeng 曾思詠 Effective Evaluation of Lupus Nephritis in Rat Podocyte Cell Culture |
author_sort |
Sih-Yong Zeng |
title |
Effective Evaluation of Lupus Nephritis in Rat Podocyte Cell Culture |
title_short |
Effective Evaluation of Lupus Nephritis in Rat Podocyte Cell Culture |
title_full |
Effective Evaluation of Lupus Nephritis in Rat Podocyte Cell Culture |
title_fullStr |
Effective Evaluation of Lupus Nephritis in Rat Podocyte Cell Culture |
title_full_unstemmed |
Effective Evaluation of Lupus Nephritis in Rat Podocyte Cell Culture |
title_sort |
effective evaluation of lupus nephritis in rat podocyte cell culture |
publishDate |
2011 |
url |
http://ndltd.ncl.edu.tw/handle/dsb99c |
work_keys_str_mv |
AT sihyongzeng effectiveevaluationoflupusnephritisinratpodocytecellculture AT céngsīyǒng effectiveevaluationoflupusnephritisinratpodocytecellculture AT sihyongzeng dàshǔshènzàngzúxìbāotàntǎolángchuāngshènyánliáoxiàopínggū AT céngsīyǒng dàshǔshènzàngzúxìbāotàntǎolángchuāngshènyánliáoxiàopínggū |
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1718625947008106496 |