Immunohistochemical staining for α-SMA showed strong positive reaction in numerous MCs (Fig. 6).
Podocytes with heterochromatic nuclei and distorted fused foot processes were detected. Irregular thickening of the GBM with lack of fenestration of glomerular capillaries was seen (Fig. 8). Lysosomes, phagolysosomes, dense bodies and vacuoles were observed in the cytoplasm of some podocytes (Figs 9 and 10). Numerous MCs with irregular heterochromatic nuclei were also observed (Fig. 10).
H&E-stained sections revealed most of the corpuscles as normal, whereas some of them were distorted. Numerous MCs (glomerular hypercellularity) were present (Fig. 11a). Positive reaction for PAS showed relatively thin basement membrane of Bowman's capsules and distributed inter-capillaries mesangial matrix (Fig. 11B). Also, positive reaction for α-SMA appeared in some MCs (Fig. 11c).
Toluidine blue-stained semithin sections showed renal glomeruli with anastomosing networks of blood capillaries. The capillary walls were lined by endothelial cells with flat nuclei. MCs and their matrix were seen between the capillaries. Bowman's capsule had a regular parietal layer of flat cells and a visceral layer of large podocytes (Fig. 12).
Ultrastructurally, some podocytes appeared with euchromatic indented nuclei and well-defined primary and secondary foot processes. The filtration slits in between the foot processes were preserved. Small mitochondria and few rough endoplasmic reticulum were seen in the cytoplasm of podocytes. Regular relatively thin GBMs were detected. Thickened areas lacking endothelial fenestrations were observed in some GBMs (Fig. 13).
Aminoglycosides have been known as highly effective antimicrobial agents for more than 50 years. They are considered to be very effective against many life-threatening infections, especially those caused by Gram-negative bacteria, as they display many highly desirable properties including low rate of true resistance, low cost of therapy and rapid concentration-dependent bactericidal effects [27,28]. Gentamicin is one of the most important and effective aminoglycoside antibiotics used widely for treatment of serious infections. Adverse effects such as nephrotoxicity constitute one of the major limitations to their use [29,30]. Several studies have shown that gentamicin causes other infections such as hepatotoxicity  and testicular toxicity .
In the current work, features of apoptosis were seen in some podocytes, which appeared small with darkly stained nuclei. Their cytoplasm contained multiple lysosomes and vacuoles. They had distorted fused foot processes. This was in accordance with other previous findings [29,40]. The researchers mentioned that cellular damages could be referred to renal ischaemia caused by toxic effect of gentamicin on renal blood vessels; they added that gentamicin leads to generation of ROS, which are important modulators of renal blood flow. Moreover, the fusion of pedicles after administration of adriamycin in rats was suggested to be a result of loss of electrical load in podocyte pedicles . Also, gentamicin chelates mitochondrial iron, forming a very oxidant Fe-II gentamicin complex capable of causing cell death and triggering the apoptotic pathway . The increased number of lysosomes found in experimental animals administered gentamicin could be explained by their uptake and accumulation of gentamicin as previously reported . In addition, the external surface of podocytes is covered with a sialic acid rich in glycocalyx known as podocalyxin. Podocalyxin is the target of glomerular diseases that affect the shape of foot processes and reduce the components of slit diaphragm with subsequent development of albuminuria .
Cellular infiltration was observed in this study. This result is in accordance with those of previous investigators [42,43], who stated that gentamicin evokes an inflammatory response in experimental animals and in humans with cell infiltration, increased cytokine production and increased capillary permeability. They added that the inflammatory response initially appears as a defense mechanism, but after that it contributes to renal damage progression.
MCs resemble smooth muscle cells; they can modulate glomerular haemodynamics by controlling the glomerular capillary surface area. Only activated proliferating cells express α-SMA in mesangial injury and that was thought to be a sign of activation . In the current work, immunoexpression for α-SMA was not detected in normal glomeruli. Also, mesangial hypercellularity was confirmed by extensive positive PAS staining of the mesangial matrix and strong positive α-SMA immunoexpression in MCs. Moreover, hypertrophy and proliferation of endothelial and MCs were recorded. This is in agreement with previous studies [2,28,44,45]. Those researchers reported that MCs secrete endothelin-1 in response to glomerular injury. Endothelin-1 acts by autocrine and paracrine mechanisms on MCs, causing their proliferation and excessive accumulation of its matrix. Hypertrophy and proliferation of MCs occurred because of the mitogenic effect of endothelin-1 and by mediating the proliferative effect of other growth factors such as angiotensin II. They added that endothelin I increased fibronectin, type IV collagen and type I collagen by MCs.
The immunohistochemical assessment of α-SMA was carried out to determine the role of MCs and glomerular myofibroblasts in the progression of gentamicin glomerular injury and to assess their value in determining long-term renal outcome. Increased expression of α-SMA in glomerulonephropathies was previously observed by many authors , who regarded this increase as a sign of mesangial activation and proliferation at sites of mesangial injury. They suggested that the possible myofibroblast migration from the interstitium to the damaged glomeruli could be associated with mesangial injury. Recently, smooth muscle protein was seen to be expressed in glomeruli during glomerulogenesis and to decrease with glomerular maturation. It is increased again in some glomerulopathies and this correlates with a worse disease prognosis as a higher rate of apoptosis and lower rate of proliferation occurred with it .
There is no conflict of interest to declare.
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