The two isoforms of matrix metalloproteinase- 2 have distinct renal spatial and temporal distributions in murine models of types 1 and 2 diabetes mellitus

Abstract

BackgroundWe recently reported on the enhanced tubular expression of two discrete isoforms of the MMP-2 (full length and N-terminal truncated, FL-MMP-2, NTT-MMP-2) in a murine model and human diabetic kidneys. In the present study, we examined in more detail the temporal and spatial distributions of MMP-2 isoform expression in murine models of Type 1 and Type 2 diabetes mellitus.MethodsDiabetic models were streptozotocin (STZ)-induced diabetes (Type 1 diabetes mellitus) and db/db mice (Type 2 diabetes mellitus). We quantified the abundance of two isoforms of MMP-2 transcripts by qPCR. A spatial distribution of two isoforms of MMP-2 was analyzed semi-quantitatively according to time after injection of STZ and with increasing age of db/db mice. Furthermore, immunohistochemistry for nitrotyrosine was performed to examine a potential association between oxidative stress and MMP-2 isoform expression.ResultsBoth isoforms of MMP-2 were upregulated in whole kidneys from STZ and db/db mice. In the case of FL-MMP-2, mRNA levels significantly increased at 12 and 24 weeks in STZ mice, while the isoform expression was significantly increased only at 16 weeks, in the db/db mice. FL-MMP-2 protein levels increased in the cortices and outer medullae of both STZ and db/db mice as a function of the duration of diabetes. For NTT-MMP-2, mRNA levels increased earlier at 4 weeks in STZ mice and at 10 weeks of age in db/db mice. The expression of NTT-MMP-2 also increased, primarily in the cortices of STZ and db/db mice, as a function of the duration of diabetes. Quantitatively, these findings were consistent with the qPCR results in the case of NTT-MMP-2, respectively (STZ 24 weeks, 3.24 ± 3.70 fold; 16 weeks db/db, 4.49 ± 0.55 fold). In addition, nitrotyrosine was expressed primarily in cortex as compared to medulla as a function of the duration of diabetes similar to NTT-MMP-2 expression.ConclusionsTwo isoforms of MMP-2 are highly inducible in two diabetic murine models and become more abundant as a function of time. As the expression patterns were not the same in the two isoforms of MMP-2, it is possible that each isoform has a discrete role in the development of diabetic renal injury.

Anodal and cathodal tDCS modulate neural activity and selectively affect GABA and glutamate syntheses in the visual cortex of cats

“…qPCR. qPCR was performed as described previously (Kim et al 2018;Rahman et al 2018). In brief, total RNA was extracted from the frozen tissue of A21a using the RNAprep Pure Tissue Kit (Tiangen Biotech, Beijing, China) and reverse transcription was performed on 2 μL of total RNA using the FastQuant RT Kit (with gDNase) (Tiangen Biotech) in accordance with the manufacturer's instructions.…”

“…The sequences of the primers used in the RT-PCR are listed in Table 1. The relative mRNA concentration of the target genes was calculated vs. the corresponding GAPDH Ct value and expressed in a 2 − Ct method (Kim et al 2018;Rahman et al 2018).…”

“…5B). One-way ANOVA comparison of the relative mRNA concentrations, as computed using the 2 − Ct method (Kim et al 2018;Rahman et al 2018), indicated that the concentration of GAD65 and GAD67 in A21a was significantly affected by the tDCS polarity (GAD65: F 2,15 = 9.204, p = 0.002; GAD67: F 2,15 = 4.575, p = 0.028). Post hoc pairwise tests The histograms with error bars (SDs) represent the mean concentration of GABA (A) and GLU (B) across six cats after s-, aand c-tDCS, respectively.…”

Matrix Metalloproteinases in Diabetic Kidney Disease

“…These changes were attenuated by renin-angiotensin system (RAS) inhibition [58]. Recently, two different isoforms of MMP-2 were identified in diabetic models showing differentially distribution in renal tissue [59]. An interesting finding on genetic susceptibility to DKD was described by Fornoni et al, who analysed mesangial cells from murine strains resistant (B6) and susceptible (ROP) to sclerosis, mimicking patients who develop DKD despite an adequate glycaemic control.…”

“…Also, different conditions and time of evolution of disease [57] are presented according to in vitro or in vivo models. Moreover, it has been reported that murine strains may also show certain discrepancies in MMPs outcomes according to the genetic background [59].…”