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Objective To investigate the effect of albumin on expression of NLRP3 inflammasome and its downstream cytokines IL-1β and IL-18 in tubular epithelial cells.Methods Thirty mesangioproliferative glomerulonephritis (MsPGN) patients with different levels of proteinuria were selected, and their renal biopsy samples were stained by PAS and Masson to observe tubular epithelial cells injury and inflammatory cells infiltration.NLRP3, caspase-1, IL-1β and IL-18, as well as different inflammatory cells, were detected by immunohistostaining.In vitro, Western blotting and real-time PCR were employed to detect NLRP3, caspase-1, IL-1β and IL-18 protein and mRNA in HK-2 cells stimulated by bovine serum albumin (BSA) (20 g/L).Results In MsPGN patients with high levels of proteinuria, there were obvious renal tubular epithelial cell injury and inflammatory cells infiltration (all P ＜ 0.05), and the expressions of NLRP3, caspase-1, IL-1β and IL-18 were up-regulated compared to patients with low levels of proteinuria (all P ＜ 0.05).Furthermore, IL-1β and IL-18expressions were positively correlated with the degree of proteinuria (r=0.836, P ＜ 0.05;r=0.901, P ＜0.05).NLRP3, caspase-1, IL-1β and IL-18 protein and mRNA were significantly increased in HK-2cells stimulated by BSA compared to the control group (all P ＜ 0.05).Conclusions Albumin is able to induce NLRP3 inflammasome activation in tubular epithelial cells, which may be the mechanism of tubulointerstitial injury and inflammation caused by proteinuria.
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Abtract:Purpose To analyze epidemiological characteristics and pathological types of 1 645 renal biopsies in Jiangsu province. Methods The reports of 1 654 percutaneous renal biopsies performed from January 2009 to June 2013 were retrospectively analysed . Results 1 597 out of 1 645 renal patients were successfully biopsied with a success rate of 97. 1%. Primary glomerular diseases ac-counted for 78. 56% of the total patients, secondary glomerular diseases 18. 71%. IgA nephropathy and mesangial proliferative lession accounted for high percent of primary glomerular diseases. Lupus nephritis was the most frequent pathologic type of secondary glomeru-lar diseases, followed by allergic purpura nephritis and diabetic nephropathy. Mesangial proliferative glomerulonephritis and hyperten-sive renal injury were more common in the Southern than in the Northern Jiangsu province, while acute tubular necrosis and allergic purpura nephritis were less in the Southern Jiangsu province. Conclusions Primary glomerular disease is still the most frequent glo-merular diseases in Jiangsu province, among which the IgA nephropathy was predominated. In secondary glomerular disease, lupus ne-phritis is the most frequent pathological type. The incidences of kidney diseases have geographical variation.
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Objective To investigate the influence of albumin on the expression of angiotensin-eonverting enzyme (ACE) in cultured human proximal tubular ceils (HK-2). Methods HK-2 cells were exposed to 2.5, 5, 10 g/L bovine serum albumin (BSA) for 6 h and 12 h respectively. The expression of ACE in HK-2 cells was detected by real-time RT-PCR and Western blot. Results Compared to the control group, the expression of HK-2 cells ACE mRNA treated for 12 h with different concentrations of BSA (2.5, 5, 10 g/L) significantly increased (P<O.05). Furthermore, Western blot analysis showed that the expression of ACE protein induced by BSA significantly increased (P<0.05). Treated with BSA (10 g/L) for 6 h and 12 h, the expression of ACE mRNA significantly increased in a time-dependent manner (P <0.05, respectively), and the ACE protein expression significantly increased (P<0.05, respectively). Conclusion BSA can induce ACE up-regulation in proximal tubular cells, which may lead to the increased production of local Ang Ⅱ and finally contributes to the intrarenal activation of renin angiotensin system (RAS).
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To obtain an ideal recombinant C-terminal fragment of the merozoite surface protein of Plasmodium falciparum in the Pichia pastoris expression system, the major surface protein-119 (MSP-119) gene sequence bearing the 6-his gene was inserted into expression vector pPIC9k and the target gene was transformed to the susceptible yeast cells GS115 by using electroporation. The multiple inserts were screened and the successfully expressed MSP-119 protein with the relative molecular weight of 12kDa in the supernatants of cell cultures could be detected by SDS-PAGE. Meanwhile, Western blot analysis also demonstrated that this protein reacted with mouse anti-MSP-119 monoclonal antibody, and the expression level of MSP-119 was more than 1.0 g/L. It is concluded that this recombinant protein expressed in the Pichia pastoris expression system resembles the native proteins existed.