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A Novel Mechanism for Renal Osmoregulation

Author: ZuoJunZuo
Tutor: YangYunQing
School: Xiamen University
Course: Biochemistry and Molecular Biology
Keywords: Triglycerides Lipid droplets Hypertonic stress
CLC: Q25
Type: Master's thesis
Year: 2009
Downloads: 30
Quote: 0
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Abstract


Cellular osmotic adjustment is one of the most basic physiological activities . However , the cells adapt to hypertonic stress conditions and protection mechanisms are still many unknown places . In the present study , we mIMCD3 cells ( murine renal medullary collecting duct cells ) as a model , using Oil Red O ( oil red ) staining , chemical measurements and flow cytometry techniques , and explore the hypertonic stress on renal effects of intracellular lipid homeostasis . We found that in mIMCD3 cells, hypertonic stress treatment (550-900 mOsmol / kg, NaCl) caused the accumulation of intracellular triglycerides increased , thus gathered a large number of intracellular lipid droplets . Optional ACC ( acetyl coenzyme A carboxylase ) and FASN ( fatty acid synthase ) of chemical inhibitors and Cerulenin accompanied TOFA 550 mOsmol / kg hypertonic cells were processed mIMCD3 24h, can be reduced to different extents hypertonic stress-induced increase of TG content , and can cause the cells in the higher osmotic stress (900 mOsmol / kg) significantly decreased survival . These results suggest that we , ACC and FASN these two key enzymes involved in fatty acid synthesis hypertonic stress increased TG accumulation part of the process . Meanwhile , we have proved that mIMCD3 cells, hypertonic stress -induced increase in TG accumulation is a protective effect. We further through the Western Blot and semi- quantitative RT-PCR method was determined and lipid metabolism -related transcription factor SREBP1 ( sterol regulatory element binding protein 1), PPARγ ( peroxisome proliferator- activated receptor γ) protein levels as well as the part of these two transcription factor target gene mRNA expression in hypertonic stress changes . To our surprise , hypertonic 700 mOsmol / kg for 24 hours after these important fat synthesis and transcription factor protein synthase mRNA levels compared with isotonic control group and no significant changes. These results may suggest that the hypertonic stress mIMCD3 cells increased TG levels may not be the appropriate level of functional genes strengthened , but is likely to involve post-transcriptional regulation and protein post-translational modification .

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