Biotechnic & Histochemistry最新文献

A diabetogenic high fat diet (HFD) can be used to induce insulin resistance and obesity in animal models; however, its effects on bone are unknown. We investigated the effects of long term HFD on bone in ovariectomized (OVX) female rats. We used 12-week-old female rats divided randomly into four groups: sham operation (sham), sham operation with HFD (SHFD), OVX and OVX with HFD (OVX + HFD). Ovaries were removed in the OVX and OVX + HFD groups and the SHFD and OVX + HFD groups were fed a HFD for 28 weeks. Serum estrogen, testosterone, lipid, adiponectin, leptin, tartrate-resistant acid phosphatase (TRAP) and N-mid fragment of osteocalcin (N-MID-OT) levels were measured. Structure, apoptosis and specific transcription factors in bone were evaluated using pathologic, densitometric and immunohistochemical analysis. Body weight, serum leptin, TRAP and testosterone levels were increased, while serum N-MID-OT, estrogen and adiponectin levels were decreased in the SHFD, OVX and OVX + HFD groups. Expression of BCL2-associated X protein, caspase-3, matrix metalloproteinase-9 and calcitonin was increased, while bone mineral density (BMD) and content (BMC) in femurs and lumbar spine, and expression of B cell lymphoma 2, type 1 collagen and osteocalcin were decreased in the bones of the SHFD, OVX and OVX + HFD groups. All indices were greatest in the OVX + HFD group and HFD produced a detrimental effect on bone in both normal and OVX rats, which may be due to increased apoptosis in bone and increased leptin and decreased adiponectin levels in serum. The effects of HFD and OVX may be synergistic.

Water hyssop (Bacopa monnieri L. Pennel) is a medicinal aquatic herb used to treat diseases in South Asia. Various regeneration protocols have been developed or modified in vitro to ensure the availability of biomass and secondary metabolites of Bacopa. We applied hydrothermally treated titanium dioxide (TiO₂) nanoparticles (NPs) (TiO₂-NPs) at different concentrations. Three explants, distal portion of half leaf (DPHL), proximal portion of half leaf (PPHL) and full leaf (FL), were used to evaluate response to TiO₂. Regeneration from the three explants in vitro was similar except for shoot length. Application of TiO₂-NPs exerted significant, but variable, effects on all parameters except percentage of shoot formation, which was 100%. Interactive effects of explant and TiO₂-NPs exhibited significant, but variable, effects on fresh weight and percentage of callus formation. All explants produced more shoots using TiO₂-NPs compared to control treatments. DPHL explants with application of 8 mg/l TiO₂ produced more shoots than controls. Similarly, FL explant treated with 2 mg/l TiO₂-NPs produced more shoots/explant than controls. All concentrations of TiO₂-NPs produced significantly longer shoots compared to controls. Increased elongation of shoots justifies use of TiO₂-NPs for propagation of plants in vitro during acclimatization. Use of TiO₂-NPs for rapid elongation of shoots ultimately fosters survival of plants.

We induced experimental nephrolithiasis in female rats using ethylene glycol (EG) and ammonium chloride (AC). We investigated the effects of carvacrol, an essential oil with antioxidant and anti-inflammatory properties, on nephrolithiasis using histopathology, immunohistochemistry and biochemistry. We used 40 female rats divided into four equal groups: control group, administered olive oil; carvacrol group, administered carvacrol in olive oil; nephrolithiasis group, administered EG and AC to induce experimental nephrolithiasis; treatment group with induced nephrolithiasis and administered carvacrol in olive oil. We observed no significant difference in crystal accumulation in the treatment group compared to the nephrolithiasis group. We found a significant reduction in hydropic degeneration of tubules and degree of inflammatory cell infiltration of intertubule areas. We also found a significant reduction in immunohistochemical staining of macrophage- and monocyte-specific antigens. Carvacrol treatment reversed the induced nephrolithiasis, increased malondialdehyde and urea, and decreased levels of glutathione peroxidase and catalase. Although carvacrol did not decrease crystal accumulation, it reduced pathological and biochemical damage, and improved kidney function by lowering the serum urea level.

Exercise training increases fibronectin type III domain-containing protein 5 (FNDC5/irisin) via the peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α)-pathway. The PGC1α pathway induced FNDC5/irisin changes in response to exercise training and ischemic stroke are not entirely understood. We investigated the relation of the PGC-1α/FNDC5/irisin pathway to exercise training and to the pathophysiology of ischemic stroke in paretic muscles of stroke-induced rat models. We induced cerebral ischemia following completion of high-intensity interval training (HIIT) to evaluate PGC1α-pathway biofactors in paretic muscles. To define the underlying molecular mechanisms for improvement in paretic muscles following cerebral ischemia, we evaluated PCG-1α-pathway factors using immunofluorescence tracking and enzyme-linked immunosorbent assay (ELISA) immunoassay. We found that HIIT for 3 weeks produced increased expression and release of PGC-1α-pathway biomarkers in both the serum and paretic muscle of stroke-induced rats. We also found a close relation between the expression of PCG-1α-pathway factors in skeletal muscle and their concentration in blood. We found that PGC-1α-pathway biomarkers cause irisin up-regulation following induction of cerebral ischemia. The reduction in neurofunctional deficits following increased PGC-1α-pathway biomarkers suggests that these factors may act as markers of improvement in paretic muscle healing following cerebral ischemia.

Propofol and dexmedetomidine (DEX) are widely used for anesthesia and sedation. We investigated the effects of propofol and DEX separately and in combination on the metabolic profile of carnitine in cultured normal human bronchial epithelial cells (BEAS-2B). Cells of the propofol group were cultured with 2 µg/ml propofol in RPMI-1640 medium. Cells of the DEX group were cultured with 0.2 ng/m DEX in RPMI-1640 medium. Cells of the propofol + DEX group were cultured with 2 μg/ml propofol + 0.2 ng/ml DEX in RPMI-1640 medium. The control group was untreated. Cells were incubated for 3 h following treatments. The effects of the drugs on cell viability were assessed using the MTT method and by microscopic examination following staining with acridine orange/ethidium bromide. The effects of drugs on carnitine, acetyl carnitine and 25 acylcarnitine derivative profiles were analyzed using liquid chromatography-tandem mass spectrophotometry. Neither propofol nor DEX affected cell viability. Administration of propofol, DEX or propofol + DEX to BEAS-2B cells caused no significant change in the concentrations of carnitine and acylcarnitine derivatives compared to the control group. We found that propofol and DEX exhibit no negative effects on the carnitine metabolism by BEAS-2B cells in vitro at clinically relevant concentrations. Our findings establish a baseline for clinical studies of the effects of propofol and DEX on carnitine metabolism.

We investigated the potential neuroprotective effects of thymoquinone (TQ) on amikacin (AK) induced oxidative damage in rat brain. We used 21 male rats divided randomly into three equal groups. The control group was injected intraperitoneally (i.p.) with 0.5 ml 0.9% aqueous NaCl and given 1 ml 0.9% aqueous NaCl orally. The AK group was administered 1.2 g/kg aqueous AK i.p. as a single dose on the day 3 of the study. The AK + TQ group was given a single 1.2 g/kg dose of AK i.p. on the day 3 of the study plus 40 mg/kg/day TQ by oral gavage daily. Treatment with TQ increased serum ferritin and decreased serum calcium levels significantly. TQ also decreased NADPH oxidase-2, NADPH oxidase-4, and caspase-3 levels. Decreased malondialdehyde (MDA) levels and increased superoxide dismutase (SOD) and catalase (CAT) activities were detected in the AK + TQ group compared to the AK group. TQ administration inhibited lipid peroxide formation and blocked oxidative reactions, which reduced the MDA level and increased SOD and CAT activities induced by AK. Oxidative damage caused by AK was ameliorated by TQ treatment owing to its antioxidative and anti-apoptotic effects. TQ may be a potential therapeutic agent for reducing the severity of AK induced oxidative damage to the brain.

An aqueous 7-amino-4-methylcoumarin (AMC) solution exhibits strong fluorescence under ultraviolet (UV) light and can be used as a Schiff reagent to visualize aldehydes. We investigated hemalum and eosin (H & E) and AMC staining for histological and pathological analysis. Sections of normal and lesioned human tissues were stained with combined H & E/AMC staining. After H & E/AMC staining, the H & E morphology was preserved under bright field microscopy. The AMC fluorescent signals observed under UV light were intense and the staining pattern was identical to that obtained by periodic acid-Schiff (PAS) staining. AMC staining of archived H & E sections also was successful. Diastase digestion differentiated glycogen from other AMC positive elements. Using H & E/AMC staining, mucus-rich adenocarcinoma cells, amebic trophozoites and fungal hyphae were visualized clearly under UV excitation. Using H & E/AMC staining, H & E and PAS-like histological imaging can be obtained using a single tissue section. H & E/AMC is useful for pathologic diagnosis especially when information from PAS staining is critical, the number of tissue sections is limited and/or the lesion in question is small.

Doxorubicin (DOX) is used as an anticancer drug despite its many side effects. Thymoquinone (THQ) is a plant-derived substance that exhibits antioxidant and anti-inflammatory properties. We investigated the protective effects of THQ on DOX induced nephrotoxicity in rats. Rats were divided into five groups of eight: group 1, untreated control; group 2, olive oil group given olive oil intraperitoneally (i.p.) for 14 days; group 3, THQ group given 10 mg/kg THQ i.p. for 14 days; group 4, DOX group given a single dose of 15 mg/kg DOX i.p. on day 7 of experiment; group 5, DOX + THQ given 10 mg/kg THQ i.p. for 14 days and 15 mg/kg DOX i.p. on day 7. Kidney tissues were evaluated for histopathology. Caspase-3, IL-17, GRP78 and TNF-α immunostaining was used to determine the expression levels of these proteins among the groups. The TUNEL method was used to determine the apoptotic index. Total antioxidant status (TAS), total oxidant status (TOS), and TNF-α and TGF-β1 levels in kidney tissue were measured using ELISA assay. Histopathologic damage, caspase-3, IL-17, GRP78 and TNF-α immunoreactivity, TUNEL positive cells, TOS, TNF-α and TGF-β1 levels were increased in group 4 compared to group 1. The TAS of group 4 decreased compared to group 1. We found decreased caspase-3, IL-17, GRP78 and TNF-α expressions and TUNEL positive cells in group 5 compared to group 4. In rats given DOX, THQ reduced kidney damage by suppressing endoplasmic reticulum stress, inflammation and apoptosis pathways.

Male reproductive dysfunction is a common complication of diabetes mellitus. Trifolium pratense exhibits antioxidant and antidiabetic effects. We investigated the effects of an extract of T. pratense on serum antioxidant status, sperm characteristics, testicular tissue changes and testosterone level in diabetic rats. Male Wistar rats were divided into six groups: 1, untreated control; 2, diabetic; 3 and 4, 100 or 200 mg/kg T. pratense extract treated, respectively; 5 and 6, diabetic 100 or 200 mg/kg T. pratense extract treated, respectively. Diabetes was induced by intraperitoneal injection of streptozotocin. After 3 weeks, serum glucose, testosterone and nitric oxide (NO); sperm parameters; testicular histology and total antioxidant capacity (TAC) were evaluated. In diabetic rats treated with T. pratense extract, sperm motility, count and viability, as well as TAC and testosterone were increased significantly compared to untreated diabetic rats, while serum NO and bcl-2 and p53 expression was decreased significantly compared to untreated diabetic rats. T. pratense extract reduced testicular tissue destruction caused by diabetes.