Biotechnic & Histochemistry最新文献

We investigated whether silibinin, a flavonoid, might be useful for treating diabetes mellitus by treating five groups of rat RINm5F β-insulinemia cells as follows: control streptozotocin (STZ) group administered citrate buffer and dimethyl sulfoxide; STZ group administered 20 mM STZ; silibinin group administered 50 µM silibinin; pre-silibinin group administered 50 µM silibinin 5 h before administering 20 mM STZ; simultaneous group administered 50 µM silibinin at the same time as 20 mM STZ. For all groups, MTT assay and flow cytometry were used to evaluate cell viability and necrosis, respectively. Glucose-stimulated insulin secretion (GSIS) and insulin cell content were determined using enzyme-linked immunosorbent assay. Also, expression of genes, pancreatic and duodenal homeobox 1 (pdx1), neuronal differentiation 1 (neurod1), v-maf avian musculoaponeurotic fibrosarcoma oncogene homolog A (mafa), glucose transporter 2 (glut2)) was determined using the real-time polymerase chain reaction. We found that silibinin improved the viability of RINm5F cells and increased GSIS and cellular insulin under glucotoxic conditions. Silibinin increased the expression of neurod1, mafa and glut2, but reduced pdx1 expression. Our findings suggest that silibinin might increase glucose sensitivity and insulin synthesis under glucotoxic conditions, which could be useful for diabetes treatment.

We investigated the effects of apocynin (APO) on experimental sciatic nerve compression injury in rabbits. We used 21 male rabbits divided randomly into three groups of seven. The control group was subjected to sciatic nerve compression with no further intervention. The APO treated group was subjected to compression injury and 20 mg/kg APO was administered daily for 21 days by intraperitoneal injection beginning the day after the injury. The sham group was treated with APO without injury. The control group exhibited shrinkage of axons, disruption of myelin sheaths and loss of nerve fibers. The damage for the control group was significantly greater than for the sham group. The severity of histopathology was decreased in the APO treated group compared to the control group, as was the oxidative stress index. Our findings suggest that APO treatment may contribute to healing of sciatic nerve damage.

Insulin receptor substrate 2 (IRS2) participates in reproduction; however, the location and expression of IRS2 in the reproductive system of female mice is not clear. We used real-time quantitative polymerase chain reaction (RT-PCR), western blot and immunohistochemical staining to investigate the expression of IRS2 in the ovary, oviduct and uterus of female mice during the estrous cycle. We found that IRS2 was expressed in all reproductive organs of mouse and that the expression level changed with the estrous phases. The expression of IRS2 in reproductive organs was greatest during estrus.

Primary or metastatic hepatic malignancies are common. Partial hepatectomy (PH) is the primary treatment for both benign and malignant hepatic neoplasms; it also is used for living donor liver transplantation. The regenerative potential of the liver after PH is 70-80% in humans. We investigated the protective and therapeutic effects of agomelatine (AGM) on rat liver regeneration following PH. We used 32 rats distributed equally into four groups: group 1, sham control; group 2, PH group; group 3, administered 20 mg/kg AGM orally once/day for 7 days following PH; group 4, administered 20 mg/kg AGM orally once/day 3 days before and 7 days following PH for 10 days. Liver samples were analyzed for antioxidants and free radicals. Tissue samples were processed and stained with hematoxylin and eosin to assess histopathological status and stained immunohistochemically for Ki-67. We found that PH reduced antioxidant enzymes and increased tissue reactive oxygen species, whereas AGM treatment had the opposite effect on these parameters. Our biochemical and histopathological findings were consistent. PH caused sinusoid congestion and dilation. Intensity of Ki-67 immunostaining of hepatocytes was increased in group 2, whereas these were reduced in group 4. Intensity of Ki-67 immunostaining of hepatocytes was increased in group 2, whereas it was reduced in the group 4 compared to group 1. We found that AGM was hepatoprotective following PH due to its antioxidant and free radical scavenger properties.

The antipsychotic drug, olanzapine, is prescribed for postpartum psychosis. Possible adverse effects on fertility of offspring are unclear. We investigated the effects of administering olanzapine via lactation on testicular development and endocrine function of prepuberal male rats. Olanzapine was administered to mothers at 2.5, 5 or 10 mg/kg. We found in male offspring increased body weight, decreased gonadosomatic index, testicular weight and epididymal weight. The volume of seminiferous tubules, seminiferous epithelium, Leydig cells, intertubule tissue and lymphatic space was reduced in rat pups exposed to olanzapine. Tubule diameter and length, seminiferous epithelium height, Leydig cell size and nuclear diameter also were reduced. Testosterone levels were reduced in the groups exposed to olanzapine, while prolactin levels were increased. We observed histopathology in testes of animals whose mothers had been treated with 2.5 mg/kg olanzapine; more severe pathology was observed in offspring whose mothers were administered higher doses. Administration of olanzapine to mothers during lactation produced testicular and endocrine pathology in prepuberal rats in a dose-dependent manner.

Bevacizumab is a recombinant humanized monoclonal antibody whose adverse effects include cardiotoxicity. We investigated whether using adenosine triphosphate (ATP) or benidipine either separately or together protects against cardiac damage induced by bevacizumab in rats. Forty Wistar albino male rats were allocated to five groups of eight: bevacizumab (Bv), ATP + bevacizumab (ABv), benidipine + bevacizumab (BBv), ATP + benidipine + bevacizumab (ABBv) and untreated controls. Rats in the ABv group were injected intraperitoneally (i.p.) with 2 mg/kg ATP. The BBv group was given 4 mg/kg benidipine by oral gavage. The ABBv group was injected i.p. with 2 mg/kg ATP and simultaneously administered 4 mg/kg benidipine orally. One hour after administration of ATP, benidipine or normal saline, the Bv, ABv, BBv and ABBv groups were injected i.p. with 10 mg/kg bevacizumab. Malondialdehyde (MDA) and total glutathione (tGSH) levels were measured in cardiac tissue, and troponin I (TP I) and creatine kinase MB (CK-MB) levels were measured in blood samples. Tissue samples were examined for histopathology. We found the lowest TP I, CK-MB and MDA levels and the highest tGSH level in the ABBv group; these results were similar to the control group. Nuclei of cardiomyocytes in the BV group were misshapen and shrunken, and myofibers were disrupted; we also observed eosinophilic degeneration and interstitial edema. Blood capillaries were dilated and congested. We observed amelioration of these findings in the ABBv group. We found that ATP and benidipine alone or in combination reduced cardiac damage associated with the use of bevacizumab. ATP + benidipine combined therapy produced the most favorable results.

The anatomy of the carotid body (CB) and its nerve supply are important, because it is a potential therapeutic target for treatment of various clinical conditions. Visualization of the CB in situ in fixed human anatomical specimens is hampered by obscuring adipose and connective tissues. We developed a tissue clearing method to optimize identification of the CB. We used single sided carotid bifurcations of six human cadavers fixed long term. Visualization of the CB was accomplished by clearing tissue with xylene. Under incident light, carotid bifurcations exhibited a less transparent, darker colored CB; hematoxylin and eosin stained paraffin sections confirmed its identity. Our visualization of the CB in situ in human carotid bifurcations fixed long term enabled targeted resection and subsequent topographic and morphometric measurements of the CB. Our procedure does not interfere with immunohistochemical staining of sections prepared from such specimens.

I investigated the effects of diazoxide, a mitochondrial potassium channel opener, on streptozotocin (STZ) induced pancreatic β cell damage via the HSP70/HSP90/TLR4/AMPK signaling pathways in vitro. I used the pancreatic β cell line, 1.1B4, to create four groups: control, STZ treated, diazoxide treated, STZ + diazoxide treated. The STZ treated cells were exposed to 20 µM STZ for 2 h with or without 100 µM diazoxide for 24 h. Total antioxidant status (TAS), total oxidant status (TOS), cell viability and mitochondrial membrane potential (MMP) were measured. Expression of ATP-sensitive potassium channel (K_ATP) subunits, heat shock protein-70 (HSP70), heat shock protein-90 (HSP90), toll-like receptor 4 (TLR4), AMP-activated protein kinase (AMPK) and some apoptotic proteins were detected using western blotting. Apoptosis was assessed using TUNEL staining. STZ increased TOS and OSI in the pancreatic β cells; however, diazoxide failed to improve oxidative stress. Also, STZ increased tunnel positive cells in the pancreatic β cells. Diazoxide decreased the tunnel positive cells in the STZ treated β cell. STZ decreased MMP; however, diazoxide did not normalize MMP in the STZ induced β cells. Diazoxide increased the HSP70:HSP90 protein expression ratio. STZ decreased expression of AMPK and subunits of K_ATP channel and increased the expression of caspase-3 and TLR4 protein; diazoxide normalized the expression of all proteins studied. K_ATP channel opening by diazoxide protects pancreatic β cells against STZ toxicity via HSP70/HSP90/TLR4/AMPK signaling.