ABSTRACT
Objective To explore the influence of fenofibrate on apoptosis of human renal proximal tubular cells(HK?2)induced by free fatty acid (FFAs). Methods Methyl azo thiazole blue(determined by MTT)was applied for detection of HK?2 cell proliferation capacity;Spectrophotometry was used to determine the expression of MDA and SOD;MCP?1 and IL?8 in culture supernatant of cells were detected by ELISA;Real?time PCR and Western blot were used to evaluate mRNA and protein expression of Bax and Bcl?2 respectively. Results FFAs inhibited HK?2 cell prolifera?tion in a dose and time dependence. mRNA and protein expression of Bax significantly increased compared with control,but mRNA and protein ex?pression of Bcl?2 decreased. After preincubation of fenofibrate,the inhibition of HK?2 cell proliferation by FFAs was alleviated;expression of SOD increased and expression of MDA,MCP?1 and IL?8 were decreased;mRNA and protein expression of Bax was significantly decreased,but mRNA and protein expression of Bcl?2 was increased. Conclusion FFAs can induce apoptosis of renal proximal tubular cell in a dose and time dependent manner. Fenofibrate can inhibite HK?2 cell apoptosis by decreasing oxidative stress,inhibiting inflammation,up?regulating expression of Bcl?2 and down?regulating expression of Bax,which alleviated nephrotoxicity of FFAs.
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AIM: To study the effect of Bene Jones protein (BJP) from multiple myeloma(MM) patient and TGF-? 1 on cultured renal proximal tubular cell(PTC) proliferation. METHODS:[ 3H] TdR incorporation was used to study the effect of ?BJP and TGF-? 1 on cultured rat NRK.52E PTC proliferation,the expression of TGF-? 1 in the supernatant of PTC cultured with BJP was assessed with ELISA. RESULTS:① [ 3H] TdR incorporation of PTC was inhibited by BJP in a dose-dependent manner,when co-cultured with 100-800 ?mol/L BJP and 2.0 ?g/L TGF-? 1, the [ 3H] TdR incorporation was lower than that of BJP alone, especially when BJP≥400 ?mol/L; ②The expression of TGF-? 1 in the supernatant of PTC cultured with BJP was increased ,especially when BJP≥400 ?mol/L( P
ABSTRACT
Objective To explore the possible mechanism of nitric oxide(NO) involved in iron-mediated cytotoxicity on renal tubular cells, meanwhile to estimate the effect of reactive oxygen sepcies scavenger on iron-mediated cytotoxicity and its relation to nitric oxide. Methods in this study, the relationship between NO production and lactate dehydrogenase(LDH) release were observed in primary subconfluent proximal tubular cells coincubated with different doses of NTA-Fe and lipopolysaccharide(LPS) alone or in combination. NO production was monitored by NO2 -- concentration in supernatant based on Griess reaction. Meanwhile, semi-quantitative RT-PCR was applied to detect the inducible nitric oxide synthase (iNOS) mRNA level induced by NTA-Fe and LPS together. In addition, experimental groups were exposed to reactive oxygen species (ROS) scavengers to determine the impact of the interaction between NO and ROS on iron-mediated cytotoxicity. Results After 12-hour coincubation, NTA-Fe could increase both LDH release and NO2 production in a dose-dependent manner (P 0. 05 ) although tubular injury was aggravated (P