Effect of progesterone administration on tissue mast cell population and histamine content in mice uterus after ovulation induction.

OBJECTIVE: Mast cell population and histamine affect on blastocyst implantation. This study aimed to evaluate the effects of progesterone administration after induction of ovulation on the uterine tissue mast cell population and histamine content in mice. METHODS: We ran an experimental study on three groups of mice; control group, ovulation induction (induction group), and ovulation induction along with progesterone administration (progesterone group). Mast cells were counted using toluidine blue staining, and the histamine level was measured through spectrophotometry. RESULTS: According to the analysis of variance (ANOVA), there was no difference in mast cell population in endometrium (p=0.138) nor in myometrium (p=0.611). The ratio of mast cells in the myometrium per endometrium increased in the progesterone group in comparison to the control group based on a generalized linear model (p=0.041). The uterine histamine level was different between the groups, based on the ANOVA (p=0.039), in which the progesterone group had lower amounts of histamine. CONCLUSIONS: Progesterone administration after ovulation induction did not decrease the number of endometrial mast cells and could have increased the ratio of myometrium mast cells per endometrium mast cell. The histamine level in uterus decreased by the administration of progesterone in the ovulation-induced mice.

Macrophage polarization in wound healing: role of aloe vera/chitosan nanohydrogel.

The balance between M1 and M2 macrophages plays an important role in wound healing. Interestingly, this immune response can be modulated by natural biomaterials such as chitosan nanohydrogel (Ch) and aloe vera (AV). Therefore, we aimed to improve wound recovery response by exploiting the potential healing properties of Ch and AV. Wounds were created in rats and were treated daily with either saline (control), AV, Ch, or different ratios of AV (volume):Ch (weight) (1:1), (2:1), and (3:1). M1 (iNOS, TNF-α) and M2 (CD163, TGF-ß) responses were analyzed at days 3, 7, 14, 21, and 28. Wound healing increased within the third and seventh days in AV-Ch (3:1) (P < 0.001 and P < 0.002, respectively). In the treated groups, immunohistochemistry of iNOS expression decreased on the third day (P < 0.0001) while CD163 increased (P < 0.0001) on the 3rd, 7th, and 14th days. The gene expression of TGF-ß decreased on the third day in AV group (P < 0.03) and on the 21st and 28th days in Ch-treated group (P < 0.00). TNF-α expression decreased in AV, Ch, and AV-Ch (3:1 v/w) on the 14th and 28th days (P < 0.00). TGF-ß and TNF-α proteins decreased on the 28th day compared to the control and AV-Ch (3:1 v/w), respectively. AV-Ch (1 and 3:1 v/w) and Ch resulted in optimum wound repair by decreasing M1 after 3 days and increasing M2 after 14. Thus, Ch nanohydrogel, especially in combination with 1:1 and 1:3 ratio to AV, could be a proper candidate for modulating macrophages in response to wound healing.

Protective effects of oleuropein against renal injury oxidative damage in alloxan-induced diabetic rats; a histological and biochemical study.

BACKGROUND: Oleuropein is a potent antioxidant and free-radical scavenger with antiinflammatory properties. OBJECTIVES: In the present study, we evaluated the protective effects of oleuropein on myeloperoxidase (MPO) activity, nitrite, urea, creatinine and glomerulosclerosis in alloxan-induced type 1 diabetic rats. MATERIALS AND METHODS: Thirty Sprague-Dawley male rats were randomly divided into 3 groups: group 1 as control; group 2 as untreated diabetic; and group 3 as treated with oleuropein 15 mg/kg i.p daily. Diabetes was induced in the second and third groups by subcutaneous alloxan injection. After 48 days, the animals were anaesthetized and then the livers and kidneys were removed immediately and used fresh or kept frozen until MPO activity analysis. Blood samples were also collected before sacrificing to measure nitrite, urea, and creatinine. Kidney paraffin sections were prepared to estimate glomerular volume, leukocyte infiltration, and glomerulosclerosis. RESULTS: Oleuropein significantly decreased leukocyte infiltration and glomerulosclerosis in the treated group compared with the diabetic untreated group. Oleuropein significantly decreased the levels of urea, nitrite, and creatinine in the treated group compared with the diabetic untreated group. Moreover, oleuropein significantly decreased MPO activity in the treated group compared with the diabetic untreated group. CONCLUSIONS: Oleuropein has antioxidative and antiatherogenic activities and exerts beneficial effects on inflammation and kidney function test and decreases diabetic complication in diabetic rats.

Stereological assessment of normal Persian squirrels (Sciurus anomalus) kidney.

The functions of the mammalian kidney are closely related to its structure. This suggests that renal function can be completely characterized by accurate knowledge of its quantitative morphological features. The aim of this study was to investigate the histomorphometric features of the kidney using design-based and unbiased stereological methods in the Persian squirrel (Sciurus anomalus), which is the only representative of the Sciuridae family in the Middle East. The left kidneys of five animals were examined. Total volume of the kidney, cortex, and medulla were determined to be 960.75 ± 87.4, 754.31 ± 77.09 and 206.1 ± 16.89 mm3, respectively. The glomerular number was 32844.03 ± 1069.19, and the total glomerular volume was estimated to be 36.7 ± 1.45 mm3. The volume and length of the proximal convoluted tubule were estimated at 585.67 ± 60.7 mm3 and 328.8 ± 14.8 m, respectively, with both values being greater than those reported in the rat kidney. The volume and length of the distal convoluted tubule were calculated at 122.34 ± 7.38 mm3 and 234.4 ± 17.45 m, respectively, which are also greater than those reported in the rat kidney. Despite the comparable body weight, the total number and mean individual volume of glomeruli in the Persian squirrel kidney were greater than those in the rat kidney. Overall, the stereological variables of the kidneys elucidated in this study are exclusive to the Persian squirrel. Our findings, together with future renal physiological data, will contribute to a better understanding of the renal structure-function relationship in the Persian squirrel.

Effect of normobaric hyperoxia on gentamicin-induced nephrotoxicity in rats.

OBJECTIVES: Gentamicin sulphate (GS) nephrotoxicity seems to be related to the generation of reactive oxygen species. There is evidence that oxygen preconditioning increases the activity of antioxidant enzymes. MATERIALS AND METHODS: Forty eight female rats were divided into 6 groups (n=8) as follows: group 1 was the control, group 2 received daily GS, groups 3,4 and 5 received oxygen 2 hr/day for 2 days, 4 hr/day for 2 days, 4 hr/day for 4 days, recpectively and then received daily GS, group 6 received oxygen 2 hr/day for 2 days and then received 2 hr oxygen before daily GS injection. Oxygen (with 90% purity) used at the flow rate of 4 l/min. GS administred for 8 days (100 mg/kg, IP). Tissue sections prepared from the left kidney, stained with PAS method and then studied hisopathologically and stereologically. The right kidneys were homogenized and the supernatants were prepared. Serum MDA, creatinine and urea, renal MDA, gluthatione and catalase activity were measured. The data were analyzed by Mann-Whitney U test at the significant level of P<0.05. RESULTS: Oxygen therapy significantly improves serum creatinine and urea, preserve tubular volume density, reduce tubular necrosis in groups 4 and 6 compared to group 2. Oxygen therapy significantly increases renal catalase in groups 4 and 6 compared to group 2. CONCLUSION: Pretreatment with normobaric hyperoxia and daily oxygen therapy improved gentamicin nephrotoxicity possibly via inhibition of lipid peroxidation and increasing the renal catalase activity but could not restore any parameter at the same levels as control group.

Amelioration of lipid peroxidation in vivo and in vitro by Satureja khozestanica essential oil in alloxan-induced diabetic rats.

BACKGROUND: We examined possible protective effect of Satureja khozestanica essential oil (SKE) on in vivo and in vitro lipid peroxidation in alloxan-induced Type 1 diabetic rats. METHODS: Thirty Sprage-dawley male rats were divided into three groups randomly; group one as control, group two diabetic untreatment, and group three treatments with SKE by 500 ppm in drinking water, respectively. Diabetes was induced in the second and third groups by alloxan injection subcutaneously. After 8 weeks, animals were anaesthetized, livers and kidneys were then removed immediately and used fresh or kept frozen until their lipid peroxidation analysis. Lipid peroxidation was determined by measurement of thiobarbituric acid reactive substances (TBARS). Blood samples were also collected before killing to measure the levels of fasting blood suger (FBS) and lipid peroxidation. RESULTS: SKE significantly inhibited the levels of FBS, TBARS serum and kidney content in treated group compared with the diabetic untreated group. Also the levels of malonedialdehyde liver content unaltered in treated group. SKE significantly inhibited LDL oxidation in vitro. CONCLUSIONS: The findings showed that SKE exerts beneficial effects on the lipid peroxidation in alloxan-induced Type 1 diabetic rats.

Effect of dimethyl sulfoxide on inhibition of post-ovariectomy osteopenia in rats.

There is increasing evidence that oxidative stress, due to estrogen deficiency, leads to osteopenia. In this study, dimethyl sulfoxide (DMSO), an antioxidant solvent, was used against post-ovariectomy osteopenia (PO) in rats. Forty female rats were divided into 5 groups randomly as follows: Sham, control group; OVX, ovariectomized group; DMSO1, ovariectomized injected DMSO (0.5 ml/kg/d ip); DMSO2, ovariectomized injected DMSO (1 ml/kg/day ip) and DMSO3, ovariectomized injected DMSO (2 ml/kg/d ip). DMSO therapy started 1 week after ovariectomy and continued for 13 weeks. After 13th weeks, sera were prepared, and then L4 vertebrae and right tibial bones rinsed in fixative. Serum bone alkaline phosphatase (BALP), osteocalcin, pyridinoline, malondialdehyde (MDA) and glutathione (GSH) were measured. Trabecular volume density, trabecular and cortex thickness were estimated. Osteoclast and osteoblast numbers were counted morphometrically. The data were analyzed by ANOVA and then post hoc Tukey test at p < 0.05. The increase of pyridinoline and decrease of BALP in DMSO injected groups were inhibited compared with OVX group (p < 0.05). In DMSO injected groups, decrease of bone density, trabecular volume density, thickness of trabecular and tibial cortex were inhibited compared with OVX group (p < 0.05). MDA decreased significantly in DMSO injected groups compared with OVX group. Osteoclast number decreased in DMSO injected groups compared with OVX group (p < 0.05). Osteoblast number did not show significant change in DMSO groups compared with OVX group. In conclusion, DMSO ameliorates PO through decrease of osteoclast number, osteoclast inhibition and osteoblast activation. These effects may probably be mediated via antioxidant property of DMSO.

Diabetic nephropathy and antioxidants.

CONTEXT: Oxidative stress has crucial role in pathogenesis of diabetic nephropathy (DN). Despite satisfactory results from antioxidant therapy in rodent, antioxidant therapy showed conflicting results in combat with DN in diabetic patients. EVIDENCE ACQUISITIONS: Directory of Open Access Journals (DOAJ), Google Scholar,Pubmed (NLM), LISTA (EBSCO) and Web of Science have been searched. RESULTS: Treatment of DN in human are insufficient with rennin angiotensin system (RAS) blockers, so additional agent ought to combine with this management. Meanwhile based on DN pathogenesis and evidences in experimental and human researches, the antioxidants are the best candidate. New multi-property antioxidants may be improved human DN that show high power antioxidant capacity, long half-life time, high permeability to mitochondrion, improve body antioxidants enzymes activity and anti-inflammatory effects. CONCLUSIONS: Based on this review and our studies on diabetic rats, rosmarinic acid a multi-property antioxidant may be useful in DN patients, but of course, needs to be proven in clinical trials studies.

Complexity of diabetic nephropathy pathogenesis and design of investigations.

Diabetic nephropathy (DN) pathogenesis is very complex and multifactorial. There are several mechanisms or pathways that hyperglycemia leads to renal injuries. Each pathway makes renal injuries via several mediators. Some mediators are common between the pathways such as reactive oxygen species (ROS) and TGF-ß and there are many overlaps and interference between the pathways. This review summarized complexity of DN pathogenesis and overlaps or interfering of mediators between the pathogenesis pathways. Besides, in the review suggested new designs of researches based on this complexity pathogenesis. The pathogenesis of DN is certainly very complex and multifactorial. From the overview of molecular mechanisms of DN pathogenesis, there are many pathways and many mediators with many interferences and overlaps between them. The focal point of this pathogenesis still unknown but it seems that RAAS system, oxidative stress and TGF-ß relatively are common between these complex tangle webs of pathogenesis.

Amelioration of altered antioxidant enzymes activity and glomerulosclerosis by coenzyme Q10 in alloxan-induced diabetic rats.

Coenzyme Q10 is a natural antioxidant and scavenging free radicals. In the present study, we examined antioxidative activities of coenzyme Q10 and possible protective effect of coenzyme Q10 on in vivo and in vitro lipid peroxidation, antioxidant enzymes activity and glomerulosclerosis in alloxan-induced type 1 diabetic rats. Thirty Sprague-Dawley male rats were divided into three groups randomly: group 1 as control, group 2 as diabetic untreatment, and group 3 as treatments with coenzyme Q10 by 15 mg/kg i.p. daily, respectively. Diabetes was induced in the second and third groups by alloxan injection subcutaneously. After 8 weeks, animals were anaesthetized, liver and kidney were then removed immediately and used fresh or kept frozen until their lipid peroxidation analysis. Blood samples were also collected before killing to measure the lipid peroxidation and antioxidant enzymes activity. Kidney paraffin sections were prepared and stained by periodic acid-Schiff method. Glomerular volume and leukocyte infiltration were estimated by stereological rules and glomerular sclerosis was studied semi-quantitatively. Coenzyme Q10 significantly inhibited leukocyte infiltration, glomerulosclerosis and the levels of malondialdehyde (MDA) serum and kidney content in treated group compared with the diabetic untreated group. Coenzyme Q10 significantly inhibited LDL oxidation in vitro. Coenzyme Q10 significantly increased the serum levels of glutathione (GSH) and serum activity of catalase (CAT) and superoxide dismutase (SOD) in treated group compared with the diabetic untreated group. Coenzyme Q10 alleviates leukocyte infiltration and glomerulosclerosis and exerts beneficial effects on the lipid peroxidation and antioxidant enzymes activity in alloxan-induced type 1 diabetic rats.

Inhibitory effect of olive leaf extract on gentamicin-induced nephrotoxicity in rats.

INTRODUCTION: Gentamicin sulphate nephrotoxicity seems to be attributed to the generation of reactive oxygen species. Olive leaf extract (OLE) has been demonstrated to have antioxidant and anti-inflammatory effects. The aim of this study was to evaluate the inhibitory effect of OLE on gentamicin-induced nephrotoxicity in rats. MATERIALS AND METHODS: Thirty-five Sprague-dawley rats were divided into 5 groups to receive saline; gentamicin, 100 mg/kg/d; and gentamicin plus OLE in 3 different doses (25 mg/kg/d, 50 mg/kg/d, and 100 mg/kg/d, once daily for 12 days. Serum and renal malondialdehyde were assessed, and tubular necrosis was studied semiquantitatively. Glomerular volume and volume density of the proximal convoluted tubules were estimated stereologically from paraffin sections. Serum creatinine and renal antioxidant enzymes activity were measured. RESULTS: Gentamicin significantly increased serum creatinine, malondialdehyde, and tubular necrosis, and decreased creatinine clearance, volume density of the proximal convoluted tubules, renal glutathione, glutathione peroxidase, catalase, and superoxide dismutase compared with the control group. Cotreatment of gentamicin and OLE significantly decreased serum creatinine, malondialdehyde, tubular necrosis, and renal malondialdehyde, and increased renal glutathione, catalase, superoxide dismutase, volume density of proximal convoluted tubules, and creatinine clearance in comparison with gentamicin-only treated group. Serum malondialdehyde, serum creatinine, tubular necrosis, and volume density of proximal convoluted tubules were maintained at the same level as that of the control group by cotreatment of gentamicin and OLE. CONCLUSIONS: Olive leaf extract ameliorates gentamicin nephrotoxicity via antioxidant activity, increase of renal glutathione content, and increase of renal antioxidant enzymes activity, except for glutathione peroxidase.

Hepatoprotective and Hypolipidemic Effects of Satureja Khuzestanica Essential Oil in Alloxan-induced Type 1 Diabetic Rats.

In the present study, we examined the antioxidative activities of Satureja khuzestanica essential oil (SKE) and possible protective effect of SKE on lipid profile, atherogenic index and liver enzyme markers in Alloxan-induced Type 1 diabetic rats. Thirty male rats were randomly divided into three groups; group one as control, group two diabetic untreatment, and group three treatments with SKE by 500 ppm in drinking water, respectively. Diabetes was induced in the second and third groups by alloxan injection subcutaneously. After 8 weeks, the levels of fasting blood glucose (FBG), triglyceride (TG), cholesterol (C), low density lipoprotein (LDL), very low density lipoprotein (VLDL), high density lipoprotein (HDL), atherogenic index and the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) of all groups were analyzed. Data were analyzed through non-parametric Man Whitney test (using SPSS 13 software) and p < 0.05 was considered significant. SKE inhibited significantly the activities of ALT and ALP and decrease FBG, TG, C, LDL and VLDL. HDL level was significantly increased when treated with the extract. The activities of AST stayed unaltered. Moreover, total antioxidant capacity of SKE was 3.20 ± 0.40 nmol of ascorbic acid equivalents/g SKE. This study showed that SKE is a source of potent antioxidants. The findings of the present study also suggest that SKE exert beneficial effects on the lipid profile, atherogenic index and liver enzymes activity in Alloxan-induced Type 1 diabetic rats.

Inhibition of gentamicin-induced renal tubular cell necrosis.

Gentamicin nephrotoxicity limit its usage against gram negative bacteria. Most researches showed that antioxidant agents improved gentamicin nephrotoxicity. According to these investigations oxidative stress play a central role in the mechanism of gentamicin induced nephrotoxicity. Recently Rafieian-Kopaei and colleagues showed that erythropoietin significantly ameliorated serum creatinine, blood urea nitrogen and tubal necrosis in gentamicin induced nephrotoxicity in rat. One of the advantages of this study is treatment of rats for 10 days by erythropoietin after inducing gentamicin nephrotoxicity and besides co- treatment of gentamicin and erythropoietin at 10 days simultaneously. They showed that erythropoietin improved significantly serum creatinine and blood urea nitrogen in gentamicin injected rats simultaneously and even after gentamicin nephrotoxicity induction. This study also showed that erythropoietin ameliorates histopathological injuries especially tubular cell necrosis that induced by gentamicin. Although the detailed renoprotective mechanisms of erythropoietin cannot be fully explained by this study but histological and biochemical results are satisfactory.

Effect of rosmarinic acid on inhibition of gentamicin induced nephrotoxicity in rats.

The present investigation reports the effect of rosmarinic acid (RA), an antioxidant on gentamicin sulphate (GS)-induced renal oxidative damage in rats. Rosmarinic acid (RA) has been demonstrated to have antioxidant, free radical scavenger and anti-inflammatory effects. Twenty-eight Sprague-Dawley rats were divided in to four equal groups as follows: group 1 (control), group 2 (GS 100 mg/kg/d ip), group 3 (GS 100 mg/kg/d ip+RA 50 mg/kg/d) and group 4 (GS 100 mg/kg/d ip+RA 100 mg/kg/d). Treatments were administrated once daily for 12 days. After 12 days 24h urine was collected, blood was sampled and kidneys were removed. Serum and kidney tissue MDA assessed by thiobarbituric acid. Kidney paraffin sections (5 µm thickness) from the left kidney stained with periodic acid Schiff. Tubular necrosis was studied semiquantitatively and glomerular volume and volume density of proximal convoluted tubule (PCT) estimated stereologically. Kidney homogenize were prepared from right kidney. Serum creatinine, urea and kidney antioxidant enzymes activity were assessed by special kits. Data were compared by SPSS 13 software and Mann-Whitney test at p < 0.05. Co treatment of GS and RA (High dose) significantly decreased serum creatinine, MDA, urea, tubular necrosis (p < 0.05) and increase renal GSH, GPX, CAT, SOD, volume density of PCT and creatinine clearance significantly in comparison with GS group (p < 0.05). Treatment with RA (high dose) maintained serum creatinine, volume density of PCT, renal GSH, GPX, SOD and MDA as the same level as control group significantly (p < 0.05). In conclusion, RA alleviates GS nephrotoxicity via antioxidant activity, increase of renal GSH content and increase of renal antioxidant enzymes activity.

Satureja khozestanica essential oil ameliorates progression of diabetic nephropathy in uninephrectomized diabetic rats.

Diabetic nephropathy is the common cause of leading to end stage of renal disease (ESRD). Satureja khozestanica essential oil (SKEO) was used as an antioxidant and antidiabetic for the inhibition of diabetic nephropathy. Forty male rats were uninephrectomized and divided in four groups randomly; group one as control, group two diabetic untreatment, groups three and four treatment with SKEO by 250 or 500 ppm in drinking water, respectively. Diabetes was induced in the second, third and fourth groups by alloxan injection subcutaneously. After eight weeks treatment, serum malondialdehyde, serum creatinine and serum urea were measured. The kidney paraffin sections were stained by periodic acid Schiff method. Glomerular volume and glomerular number were estimated by stereological rules. Glomerular sclerosis was studied semi-quantitatively. The means were compared by SPSS 13 software and Mann-Whitney test at p<0.05. Satureja khozestanica essential oil (250 or 500 ppm) significantly inhibited the progression of glomerular hypertrophy, glomerular number loss, glomerulosclerosis, lipid peroxidation, serum urea and creatinine compared with the diabetic untreated group. The level of glomerular number, serum malondialdehyde, serum creatinine and urea in the treated groups was significantly maintained at the same level as that of the control group. In conclusion, satureja essential oil significantly can ameliorate glomerular hypertrophy, loss of glomerular number, glomerulosclerosis and attenuated serum urea and serum creatinine in diabetic rats.

Effects of epigallocatechin gallate on tissue protection and functional recovery after contusive spinal cord injury in rats.

Recent studies revealed the neuroprotective effects of epigallocatechin gallate (EGCG) on a variety of neural injury .The purpose of this study was to determine the effects of EGCG on the tissue protection and behavioral improvement after spinal cord injury (SCI). Rats were randomly divided into four groups of 18 rats each as follows: sham-operated group, trauma group, and EGCG treatment groups (50 mg/kg, i.p., immediately and 1 hour after SCI). Spinal cord samples were taken 24 hours after injury and studied for determination of malodialdehyde (MDA) levels, immunohistochemistry of Bax and Bcl-2, and TUNEL reaction. Behavioral testing was performed weekly up to 6 weeks post-injury. Then, the rats were euthanized for histopathological assessment. The results showed that MDA levels were significantly decreased in EGCG treatment groups. Greater Bcl-2 and attenuated Bax expression could be detected in the EGCG-treated rats. EGCG significantly reduced TUNEL-positive rate. Also, EGCG significantly reduced the percentage of lesion area and improved behavioral function than the trauma group. On the basis of these findings, we propose that EGCG may be effective in protecting rat spinal cord from secondary injury.