To examine the effect of different concentrations of aristolochic acid I (AAI) in inducing apoptosis of cultured porcine renal cell line LLC PK1 and to investigate the relationship between intracellular free calcium ...To examine the effect of different concentrations of aristolochic acid I (AAI) in inducing apoptosis of cultured porcine renal cell line LLC PK1 and to investigate the relationship between intracellular free calcium concentration ([Ca ++ ]i) and LLC PK1 apoptosis induced by AAI and the influence of a calcium antagonist, lacidipine on apoptosis and [Ca ++ ]i Methods LLC PK1 cells were treated in different groups: a the normal group without treatment; b the group with AAI alone (0 01?g·L 1 , 0 02?g·L 1 , 0 04 ?g·L 1 , 0 08?g·L 1 ); c the group with lacidipine alone (10?ng·L 1 , 10 2 ?ng·L 1 , 10 3?ng·L 1 ); d the group with AAI (0 04?g·L 1 ) plus lacidipine (10?ng·L 1 , 10 2? ng·L 1 , 10 3?ng·L 1 ) Light microscopy, agarose gel electrophoresis, Annexin V Flous apoptosis detection kit and flow cytometry using propidium iodide staining to identify or quantify the apoptosis of LLC PK1 cells Mean [Ca ++ ]i was measured by laser confocus microscopy using Fluo 3/AM staining Results A series of morphologic changes that were characteristic of apoptosis, Annexin V Flous staining positive apoptotic cells and “DNA ladder' were identified in AAI (0 02?g·L 1 -0 08?g·L 1 ) treated LLC PK1 cells Quantitative analysis of apoptotic cells showed that the percentage of apoptotic cells in AAI (0 02?g·L 1 , 0 04?g·L 1 or 0 08?g·L 1 ) group was significantly higher than that in normal group (5 3%, 48 5%, 78 7% vs 2 6%, P <0 001) Mean [Ca ++ ]i was significantly higher in cells treated with AAI (0 04?g·L 1 ) than that in normal cells (58 01±18 89 vs 22 66±4 78, P <0 001) In group treated with AAI plus lacidipine (102?ng·L 1 , 103?ng·L 1 ), mean [Ca ++ ]i was significantly lower than that treated with AAI alone (35 47±12 85, 28 55±10 16 vs 58 01±18 89, P <0 001) And the percentage of apoptotic ce展开更多
Objective: To study the renal toxic effect of titanium dioxide nanoparticles(TiNPs)prepared by chemical and green route.Methods: TiNPs were prepared by chemical(sol gel technique) and green route(using aqueous extract...Objective: To study the renal toxic effect of titanium dioxide nanoparticles(TiNPs)prepared by chemical and green route.Methods: TiNPs were prepared by chemical(sol gel technique) and green route(using aqueous extract of Desmodium gangeticum root by using titanium tetraisopropoxide as precursor). Thus prepared TiNPs were characterized using UV–visible spectrophotometry, X-ray diffractometry and evaluated its renal toxic impact in different experimental models viz., Wistar rats(100 mg/kg b.wt.; oral), LLC-PK1 cells(100 mg/m L) and isolated renal mitochondria(0.25, 0.5 and 1 mg/m L).Results: Compared to the chemically synthesized TiNPs, Desmodium gangeticum synthesized nanoparticles showed less nephrotoxicity, determined by elevated serum renal markers like urea(62%), creatinine(35%), aspartate aminotransferase(61%) and alanine transaminase(37%) and the result was in agreement with cellular toxicity(measured by MTT assay and lactate dehydrogenase activity). Further toxicity evaluation at the level of mitochondria showed not much significant difference in TiNPs effect between two synthetic routes.Conclusions: The biochemical findings in renal tissue and epithelial cell(LLC-PK1)supported by histopathology examination and isolated mitochondrial activity showed minor toxicity with TiNPs prepared by green route(Ti NP DG) than TiNP Chem.展开更多
文摘To examine the effect of different concentrations of aristolochic acid I (AAI) in inducing apoptosis of cultured porcine renal cell line LLC PK1 and to investigate the relationship between intracellular free calcium concentration ([Ca ++ ]i) and LLC PK1 apoptosis induced by AAI and the influence of a calcium antagonist, lacidipine on apoptosis and [Ca ++ ]i Methods LLC PK1 cells were treated in different groups: a the normal group without treatment; b the group with AAI alone (0 01?g·L 1 , 0 02?g·L 1 , 0 04 ?g·L 1 , 0 08?g·L 1 ); c the group with lacidipine alone (10?ng·L 1 , 10 2 ?ng·L 1 , 10 3?ng·L 1 ); d the group with AAI (0 04?g·L 1 ) plus lacidipine (10?ng·L 1 , 10 2? ng·L 1 , 10 3?ng·L 1 ) Light microscopy, agarose gel electrophoresis, Annexin V Flous apoptosis detection kit and flow cytometry using propidium iodide staining to identify or quantify the apoptosis of LLC PK1 cells Mean [Ca ++ ]i was measured by laser confocus microscopy using Fluo 3/AM staining Results A series of morphologic changes that were characteristic of apoptosis, Annexin V Flous staining positive apoptotic cells and “DNA ladder' were identified in AAI (0 02?g·L 1 -0 08?g·L 1 ) treated LLC PK1 cells Quantitative analysis of apoptotic cells showed that the percentage of apoptotic cells in AAI (0 02?g·L 1 , 0 04?g·L 1 or 0 08?g·L 1 ) group was significantly higher than that in normal group (5 3%, 48 5%, 78 7% vs 2 6%, P <0 001) Mean [Ca ++ ]i was significantly higher in cells treated with AAI (0 04?g·L 1 ) than that in normal cells (58 01±18 89 vs 22 66±4 78, P <0 001) In group treated with AAI plus lacidipine (102?ng·L 1 , 103?ng·L 1 ), mean [Ca ++ ]i was significantly lower than that treated with AAI alone (35 47±12 85, 28 55±10 16 vs 58 01±18 89, P <0 001) And the percentage of apoptotic ce
基金partly supported by grants from the Department of Science and Technology (INSPIRE), New Delhi, India (No: DST/INSPIRE Fellowship/2013 IF130406)
文摘Objective: To study the renal toxic effect of titanium dioxide nanoparticles(TiNPs)prepared by chemical and green route.Methods: TiNPs were prepared by chemical(sol gel technique) and green route(using aqueous extract of Desmodium gangeticum root by using titanium tetraisopropoxide as precursor). Thus prepared TiNPs were characterized using UV–visible spectrophotometry, X-ray diffractometry and evaluated its renal toxic impact in different experimental models viz., Wistar rats(100 mg/kg b.wt.; oral), LLC-PK1 cells(100 mg/m L) and isolated renal mitochondria(0.25, 0.5 and 1 mg/m L).Results: Compared to the chemically synthesized TiNPs, Desmodium gangeticum synthesized nanoparticles showed less nephrotoxicity, determined by elevated serum renal markers like urea(62%), creatinine(35%), aspartate aminotransferase(61%) and alanine transaminase(37%) and the result was in agreement with cellular toxicity(measured by MTT assay and lactate dehydrogenase activity). Further toxicity evaluation at the level of mitochondria showed not much significant difference in TiNPs effect between two synthetic routes.Conclusions: The biochemical findings in renal tissue and epithelial cell(LLC-PK1)supported by histopathology examination and isolated mitochondrial activity showed minor toxicity with TiNPs prepared by green route(Ti NP DG) than TiNP Chem.
