Protective Effects Of Different Concentrations Research Articles

Exploring Nitric Oxide as a Regulator in Salt Tolerance: Insights into Photosynthetic Efficiency in Maize.

The growing issue of salinity is a significant threat to global agriculture, affecting diverse regions worldwide. Nitric oxide (NO) serves as an essential signal molecule in regulating photosynthetic performance under physiological and stress conditions. The present study reveals the protective effects of different concentrations (0-300 µM) of sodium nitroprusside (SNP, a donor of NO) on the functions of the main complexes within the photosynthetic apparatus of maize (Zea mays L. Kerala) under salt stress (150 mM NaCl). The data showed that SNP alleviates salt-induced oxidative stress and prevents changes in the fluidity of thylakoid membranes (Laurdan GP) and energy redistribution between the two photosystems (77K chlorophyll fluorescence ratio F735/F685). Chlorophyll fluorescence measurements demonstrated that the foliar spray with SNP under salt stress prevents the decline of photosystem II (PSII) open reaction centers (qP) and improves their efficiency (Φexc), thereby influencing QA- reoxidation. The data also revealed that SNP protects the rate constants for two pathways of QA- reoxidation (k1 and k2) from the changes caused by NaCl treatment alone. Additionally, there is a predominance of QA- interaction with plastoquinone in comparison to the recombination of electrons in QA QB- with the oxygen-evolving complex (OEC). The analysis of flash oxygen evolution showed that SNP treatment prevents a salt-induced 10% increase in PSII centers in the S0 state, i.e., protects the initial S0-S1 state distribution, and the modification of the Mn cluster in the OEC. Moreover, this study demonstrates that SNP-induced defense occurs on both the donor and acceptor sides of the PSII, leading to the protection of overall photosystems performance (PIABS) and efficient electron transfer from the PSII donor side to the reduction of PSI end electron acceptors (PItotal). This study clearly shows that the optimal protection under salt stress occurs at approximately 50-63 nmoles NO/g FW in leaves, corresponding to foliar spray with 50-150 µM SNP.

Role of mitochondrial autophagy and the curative effect of rehmannia glutinosa leaves total glycoside capsules on nucleos(t)ide drug-induced renal injury

Objective: To study the curative effect of rehmannia glutinosa leaves total glycoside capsules and the role of mitochondrial autophagy on nucleos(t)ide drug-induced renal injury. Methods: Adefovir dipivoxil (ADV) was used to construct a hepatitis B virus (HBV) transgenic mouse model for renal injury. Renal function was measured in each group at one and two weeks of modeling. Mitochondrial autophagy indicators were measured at two weeks of modeling in renal tissue. Transmission electron microscopy was used to detect mitochondrial autophagy phenomena in renal tissue. The model was established for two weeks. Mouse with renal injury were treated with rehmannia glutinosa leaves total glycoside capsules or isotonic saline for eight weeks by intragastric administration. Renal function was measured. Renal tissue morphology was observed. Mitochondrial autophagy indicators were detected in renal tissue. The protective effect of different concentrations of verbascoside (the main active ingredient of rehmannia glutinosa capsule) was observed on HK-2 cell damage induced by ADV. HK-2 cells were divided into control, ADV, and ADV plus verbascoside groups. The effects of verbascoside at different times and concentrations were observed on the HK-2 mitochondrial autophagy indicators. Fifty patients with chronic hepatitis B were collected who presented with renal injury after treatment with nucleos(t)ide analogs. The random number method was used to divide 29 cases into a control group that received conventional treatment. The treatment group of 21 cases was treated with rehmannia glutinosa leaves total glycoside capsules on the basis of the control group. Serum creatinine (Scr) and urinary protein were detected at eight weeks.The χ(2) test or t-test was used for statistical analysis. Results: Compared with the control group, two weeks of modeling in the ADV group induced renal function injury in HBV mice. The expression of autophagy indicators was higher in the renal tissue of the ADV group than that of the control group. Transmission electron microscopy had revealed mitochondrial autophagy in the renal tissue of the ADV group. Compared with the control group, the renal function of HBV mice treated with rehmannia glutinosa leaves total glycoside capsules improved for two months, and the expressions of autophagy indicators were down-regulated.Verbascoside promoted proliferation in ADV-damaged HK-2 cells, and the expression of autophagy indicators was down-regulated compared with the ADV alone group. In 50 patients with renal function injury, the urinary protein improvement was significantly superior in the treatment group than that in the control group, with eighteen and three cases being effective and ineffective in the treatment group and 12 and 17 cases being effective and ineffective in the control group, with a statistically significant difference (χ(2) = 9.975 0, P = 0.001 6). Serum creatinine was decreased in the treatment group compared with the control group, with 11 and 10 cases being effective and ineffective in the treatment group and 12 and 17 cases being effective and ineffective in the control group, with no statistically significant difference (χ(2) = 0.593 5, P = 0.441 1). Conclusion: Rehmannia glutinosa leaves total glycoside capsule can improve the nucleos(t)ide drug-induced renal function injury in chronic hepatitis B, possibly playing a role via inhibiting PINK1/Parkin-mediated mitochondrial autophagy.

Hydroxysafflor Yellow A Inhibits Pyroptosis and Protecting HUVECs from OGD/R via NLRP3/Caspase-1/GSDMD Pathway.

To observe the protective effect and mechanism of hydroxyl safflower yellow A (HSYA) from myocardial ischemia-reperfusion injury on human umbilical vein endothelial cells (HUVECs). HUVECs were treated with oxygen-glucose deprivation reperfusion (OGD/R) to simulate the ischemia reperfusion model, and cell counting kit-8 was used to detect the protective effect of different concentrations (1.25-160 µ mol/L) of HSYA on HUVECs after OGD/R. HSYA 80 µ mol/L was used for follow-up experiments. The contents of inflammatory cytokines interleukin (IL)-18, IL-1 β, monocyte chemotactic protein 1 (MCP-1), tumor necrosis factor α (TNF-α) and IL-6 before and after administration were measured by enzyme-linked immunosorbent assay. The protein expressions of toll-like receptor, NOD-like receptor containing pyrin domain 3 (NLRP3), gasdermin D (GSDMD) and GSDMD-N-terminal domain (GSDMD-N) before and after administration were detected by Western blot. NLRP3 inflammasome inhibitor cytokine release inhibitory drug 3 sodium salt (CRID3 sodium salt, also known as MCC950) and agonist were added, and the changes of NLRP3, cysteine-aspartic acid protease 1 (Caspase-1), GSDMD and GSDMD-N protein expressions were detected by Western blot. HSYA inhibited OGD/R-induced inflammation and significantly decreased the contents of inflammatory cytokines IL-18, IL-1 β, MCP-1, TNF-α and IL-6 (P<0.01 or P<0.05). At the same time, by inhibiting NLRP3/Caspase-1/GSDMD pathway, HSYA can reduce the occurrence of pyroptosis after OGD/R and reduce the expression of NLRP3, Caspase-1, GSDMD and GSDMD-N proteins (P<0.01). The protective effect of HSYA on HUVECs after OGD/R is related to down-regulating the expression of NLRP3 inflammasome and inhibiting pyroptosis.

The effect of different concentrations of laminarin on the quality of cryopreserved ram semen

BACKGROUND: Increasingly, sheep breeders are using artificial insemination to produce lambs, so finding methods that preserve ram sperm can be useful. OBJECTIVE: To determine the protective effects of different concentrations of laminarin on ram sperm motility, viability, abnormalities, membrane, and DNA integrity, superoxide dismutase enzyme (SOD) activity, and malondialdehyde (MDA) production after freeze-thawing. MATERIALS AND METHODS: The ejaculates of four rams were collected and stored at 35 °C. Semen samples were diluted with a tris-base extender containing 100, 200, 400, and 800 μg/mL of laminarin and a control extender containing no laminarin, then frozen in liquid nitrogen after 4 h in the refrigerator. RESULTS: In the treatment of frozen-thawed spermatozoa with 800 μg/mL laminarin, motility, viability, membrane integrity, and DNA integrity were significantly higher than in the control. In spermatozoa that were exposed to 800 μg/mL laminarin after thawing, MDA production was significantly lower than in the control group. The percentage of abnormal spermatozoa in 800 μg/mL laminarin was significantly lower than that in the control. CONCLUSION: The addition of 800 μg/mL laminarin to the freezing extender increases motility, viability, SOD activity, and plasma membrane integrity, while reducing abnormality and MDA production in freeze-thawed ram semen.

α-tocopherol stabilization by soybean oil and glyceryl monostearate made oleogel: Dynamic changes and characterization for food application

α-tocopherol loaded soybean oil and monostearin-made oleogel were fabricated to explore the protective effect of different concentrations of monostearin as an oleogelator in preserving α-tocopherol. Five oleogel samples were prepared and analyzed after 12 h incubation under 4 °C. The prepared oleogel's physicochemical characteristics were analyzed from thermos-dynamic, microstructural, oxidative, and textural perspectives. The optical microscopy and Cryo-SEM results revealed the presence and formation of needle-like crystals and lamella layers, which the XRD propounds the oleogel's stabilization by the monostearin crystals' network. The oil binding capacity (OBC), gel strength, and whiteness of the prepared oleogels improved (P < 0.05) as monostearin concentration increased. The thermo-rheological analysis also indicated robust pseudo-plastic oleogels with shear thinning behavior. Additionally, the 15-day light stability analysis of α-tocopherol content in the oleogels ameliorated significantly as the monostearin concentration increased. The results are useful in formulating α-tocopherol loaded oleogels for food applications and nutraceutical delivery in which the monostearin content can be adjusted as desired.

In vitro evaluation of sodium hyaluronate protective effect against benzalkonium chloride toxicity

AbstractPurpose: To evaluate the protective effect of different concentrations of sodium hyaluronate (SH) on the Benzalkonium chloride (BAK)‐induced toxicity using an in vitro model.Methods: The NAV14 cell line (SV40‐immortalized murine conjunctival epithelium) was used. Cell monolayers were exposed to different combinations of BAK (0.001%; 0.005%; 0.01%) and SH (0.2%; 0.3%; 0.4%) for 15 minutes; then cells were washed, and fresh culture media was added. Cell viability was evaluated after 2 h by resazurin reduction and lactate dehydrogenase (LDH) release. Also, cell migration and proliferation over 24 hours were determined by the scratch wound‐healing assay. Data were analysed by two‐way ANOVA and are shown as mean ± SD of two independent experiment with 4–6 replicates each.Results: BAK induced a concentration‐dependent decrease on cell viability (BAK 0.001%: 91 ± 14%, BAK 0.005%: 45 ± 9% and BAK 0.01%: 22 ± 10% of control cells, p &lt; 0.001) and an increase in LDH release (no BAK:0.22 ± 0.02, BAK 0.001%: 0.31 ± 0.02, BAK 0.005%: 1.14 ± 0.05 and BAK 0.01%: 1.21 ± 0.05, p &lt; 0.001). Conversely, SH neutralized these effects also in a concentration‐dependent manner (p &lt; 0.001). In the presence of SH 0.4% (highest effect), cell viability was BAK 0.001%: 104 ± 22%, BAK 0.005%: 109 ± 9% and BAK 0.01%: 75 ± 13% of control cells (p &lt; 0.001 for BAK 0.005–0.01%) while LDH release was no BAK: 0.24 ± 0.03, BAK 0.001%: 0.26 ± 0.01, BAK 0.005%: 0.37 ± 0.02 and BAK 0.01%: 0.49 ± 0.22, (vs. no SH: p &lt; 0.001 for BAK 0.005–0.01%). BAK also reduced wound closure in vitro (after 24 h, no BAK: 76 ± 14%, BAK 0.001%: 42 ± 12%, BAK 0.005%: 17 ± 16% and BAK 0.01%: 0.0% wound closure, p &lt; 0.001 for all BAK). Conversely, SH neutralized this effect in a concentration‐dependent fashion (p &lt; 0.001). In the presence of SH 0.4% (highest effect), wound closure at 24 h was: no BAK: 81 ± 15%, BAK 0.001%: 58 ± 6%, BAK 0.005%: 63 ± 10%, BAK 0.01%: 60 ± 8% (vs no SH: p &lt; 0.001 for BAK 0.005–0.01%).Conclusions: Sodium hyaluronate neutralized BAK toxicity on conjunctival epithelial cells in a concentration‐dependent manner. SH 0.4% was even protective at the highest preservative concentrations. These findings support the use of SH to mitigate BAK toxicity in long‐term antiglaucomatous treatment, although more studies are needed.

Aspergillus awamori attenuates ochratoxin A-induced renal and cardiac injuries in rabbits by activating the Nrf2/HO-1 signaling pathway and downregulating IL1β, TNFα, and iNOS gene expressions.

Ochratoxin A (OTA) is one of the most dangerous and that pollute agricultural products, inducing a variety of toxic effects in humans and animals. The current study explored the protective effect of different concentrations of Aspergillus awamori (A. awamori) against OTA (0.3 mg/kg diet) induced renal and cardiac damage by exploring its mechanism of action in 60 New Zealand white male rabbits. Dietary supplementation of A. awamori at the selected doses of 50, 100, and 150 mg/kg diet, respectively, for 2 months significantly improved the rabbit’s growth performance; modulated the suppressed immune response and restored the altered hematological parameters; reduced the elevated levels of renal injury biomarkers such as urea, creatinine, and alkaline phosphatase; and increased serum total proteins concentrations. Moreover, it also declined enzymatic activities of cardiac injury biomarkers, including AST, LDH, and CK-MB. A. awamori alleviated OTA-induced degenerative and necrotic changes in the kidney and heart of rabbits. Interestingly, A. awamori upregulated Nrf2/OH-1 signaling pathway. Therefore enhanced TAC, CAT, and SOD enzyme activities and reduced OTA-induced oxidative and nitrosative stress by declining iNOS gene expression and consequently lowered MDA and NO levels. In addition to attenuating renal and cardiac inflammation via reducing IL-1β, TNF-α gene expressions in a dose-dependent response. In conclusion,this is the first report to pinpoint that dietary incorporation of A. awamori counteracted OTA-induced renal and cardiac damage by potentiating the rabbit’s antioxidant defense system through its potent antioxidant, free radical scavenging, and anti-inflammatory properties in a dose-dependent response. Based on our observations, A. awamori could be utilized as a natural protective agent against ochratoxicosis in rabbits.Graphical abstract

Zhitong Jiangu decoction mitigates osteoarthritis in rabbits via regulation of NF-κB signaling pathway

&#x0D; &#x0D; &#x0D; &#x0D; Purpose: To investigate the effect of Zhitong Jiangu decoction (ZJD) on osteoarthritis rabbits, and the mechanism of action involved.&#x0D; Methods: Chondrocytes were obtained from the knees of osteoarthritic rabbits. These chondrocytes were randomly assigned to 7 groups: sham, 5, 10 and 20 % normal serum groups; 5 % ZJD, 10 % ZJD and 20 % ZJD groups. The gross and histopathological features of the rabbit cartilage were examined by microscopy. Each group was treated with a different concentration of rabbit normal serum or rabbit drug-containing serum. The protective effect of different concentrations of ZJD on the cells were determined. Cell proliferation and concentrations of IL-1 and MMP-3 were determined using cell counting kit (CCK) 8 and enzyme-linked immunosorbent assay (ELISA), respectively. The mRNA and protein expressions of inhibitor of nuclear factor-κB kinase α (IKK-α) and nuclear factor kappa B p65 (NF- κB p65) were determined by quantitative reverse transcription polymerase chain reaction (qRT- PCR) and immunoblotting, as appropriate.&#x0D; Results: Gross and histopathological examinations of rabbit cartilage showed that osteoarthritis was successfully established in rabbit knee joint. Cell proliferation significantly and time- and concentration-dependently increased in drug-containing serum groups, relative to sham and normal serum-containing groups. However, expressions of NF-κB p65, IL-1, MMP-3 and IKK-α were markedly and time- and concentration-dependently reduced in drug-containing serum groups, relative to sham and normal serum-containing groups (p &lt; 0.05).&#x0D; Conclusion: These results indicate that ZJD mitigates osteoarthritis in rabbits via regulation of NF-κB signaling pathway. Thus, it can potentially be developed for the management of osteoarthritis.&#x0D; &#x0D; &#x0D; &#x0D;

Protective effects of different concentrations of selenium nanoparticles on rainbow trout (Oncorhynchus mykiss) primary hepatocytes under heat stress

Heat stress leads to altered expression of associated heat shock proteins (HSPs), which are critical molecular chaperones related to cellular function in living organisms. Selenium nanoparticles (SeNPs), a nanocomposite form of Se, have a protective effect against heat stress-induced cellular damage. In this study, primary rainbow trout hepatocytes were isolated to identify the protective function of SeNPs in rainbow trout hepatocytes. Experiments were divided into five groups and SeNPs were added at concentrations of 0, 2.0, 3.0, 5.0 and 8.0 μg/mL and incubated at 18 ℃ for 4, 8, 12, 24 and 48 h respectively. Hepatocyte viability, GSH-Px and SOD activity were enhanced and MDA content was reduced following the addition of SeNPs. Expression of GSH-P1 and genes related to HSPs (including HSP70a, HSP60, HSP90β, HSP10 and HSP47) were significantly increased and the optimal concentration of SeNPs for adding to hepatocytes was identified as 5.0 µg/mL. Adding 5.0 µg/mL SeNPs following heat stress (24 ℃) increased hepatocyte viability, GSH-Px and SOD activity, while MDA levels first decreased and then increased. Expression of GSH-P1 and genes related to HSPs (including HSP70a, HSP60, HSP90β, HSP10 and HSP47) were significantly higher than controls. In summary, SeNPs and slight heat stress synergistically enhanced the expression of GSH-P1 and HSPs and protected hepatocytes from heat stress damage, suggesting that SeNPs is a potential hepatocyte protective therapeutic agent.

Everolimus attenuates glutamate-induced PC12 cells death

Background Glutamate-induced neuronal cell death plays a key role in neurodegenerative diseases such as Alzheimer’s and Parkinson’s diseases. Some recent studies reported the potential immunomodulatory and neuroprotective properties of inhibitors of serine-threonine kinase, mTOR (mammalian target of rapamycin). However, no study was conducted about the neuroprotective potential of everolimus (EVR), a selective and potent mTOR inhibitor. Therefore, this study was planned to investigate whether EVR has protective effects against glutamate-induced toxicity in PC12 cells, which are used as model for neurons injury, and to elucidate the underlying mechanism. Methods PC12 cells were concurrently treated with glutamate (8 mM) and EVR (0-40 nM) for 24 h. Then, the cells viability, apoptosis rate, and apoptosis-related proteins (caspase-3, bax and bcl-2) were measured using MTT, annexin V/PI and immunoblotting assays. Results Analyzing the protective effect of different concentrations of EVR (0-40 nM) against glutamate-induced cytotoxicity revealed a significant increase in cell viability in co-treatment regimen (p < 0.01). Also, EVR (40 nM) significantly (p < 0.01) inhibited glutamate-induced apoptosis through depressing the elevation of bax/bcl-2 ratio and expression of cleaved caspase-3, concentration depend. Conclusion The results demonstrated, for the first time, that EVR could protect against glutamate-mediated PC12 cell death via inhibiting apoptosis.

6-Gingerol Pretreatment Alleviates Hypoxia/Reoxygenation-induced H9C2 Cardiomyocyte Injury by Inhibiting Oxidative Stress and Inflammation

To observe the effect of 6-gingerol (6-G) pretreatment on hypoxia/reoxygenation (H/R) induced injury in H9C2 myocardial cell and investigate its related mechanism. The H/R in vitro model of cardiomyocytes was prepared by conventional methods. In detail, H9C2 cells were added with the nitrogen-saturated hypoxic liquid, and placed in an incubator, mixed with gas (1% O 2, 5% CO 2, 94% N 2) applying for 15 min. After culturing for 3 h, the cells were taken out and placed in an incubator (37℃, 5% CO 2) for 1 h. Before establishing the cell model, the cells were pretreated with 6-G, and the cell viability was measured by MTT method to observe the protective effect of different concentrations of 6-G on H/R-induced cell damage. The 6-G mass concentration for pretreatment that led to the highest cell viability was used for follow-up experiments. DCFH-DA fluorescent probe was used to detect the effect of 6-G pretreatment on H9C2 oxidative stress level, and the intracellular oxidative stress was observed with fluorescence microscope and flow cytometry. Western blot method was used to detect the expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in H/R-induced cell inflammatory responses. Compared with the H/R group, the cell viability of the 6-G+H/R group began to increase when the concentration of 6-G promoted to50 μg/mL. The cell viability was the highest after pretreated with 200 μg/mL 6-G. Therefore, 200 μg/mL was considered as the best 6-G intervention concentration for subsequent experiment. The content of reactive oxygen species (ROS) in the 200 μg/mL 6-G group had no significant changes compared with the control group (P>0.05), and the ROS fluorescence peak did not migrate significantly. However the ROS content in the H/R group increased significantly compared with the control (P<0.05), and the ROS fluorescence peak shifted to the right. Compared with the H/R group, the ROS content of the 6-G+H/R group decreased (P<0.05), and the ROS fluorescence peak shifted to the left. Compared with the control group, the expressions of TNF-α, IL-6, IL-1β in the 6-G group had no significant changes (P>0.05); the expressions of TNF-α, IL-6, IL-1β in the H/R group increased (P<0.05). Compared with H/R group, the expressions of TNF-α, IL-6 and IL-1β in 6-G+H/R group decreased (P<0.05). 6-G pretreatment can alleviate H/R-induced H9C2 myocardial injury, which may be related to the inhibition of oxidative stress and inflammatory responses.

Low-dose chloroquine mediated neuroprotection against n-methyl-d-aspartate induced excitotoxicity in adult mice

Objective To investigate the protective effects of different concentrations of chloroquine on RGC in n-methyl-d-aspartate (NMDA) injured mice and its possible mechanisms. Methods Fifty-four healthy male C57/BL6 mice were randomly divided into three groups, 18 in each group. The mice in low-dose chloroquine group were intraperitoneally injected with chloroquine solution at a dose of 10 mg/kg daily. Mice in high-dose chloroquine group were intraperitoneally injected with chloroquine solution at a dose of 100 mg/kg, and the mice in control group were intraperitoneally injected with the same volume of PBS. NMDA intravitreal injection was performed 2 days after intraperitoneal injection, 5 nmoles NMDA was injected into the left eye, and the same volume of PBS was injected into the right eye as a control. The RGC staining of retinal plaques were performed 7 days after NMDA injection, and the number of alive RGC was calculated. The visual acuity and electroretinogram were used to evaluate the electrophysiological functions of RGC at 9 and 10 days after modeling. Real-time quantitative PCR and retinal frozen sections and glial fibrillary acidic protein (GFAP) immunofluorescence staining were performed 11 days after NMDA injection to evaluate the glial activation of the retina. The density, visual acuity, and the amplitude of PhNR-wave of RGC between groups were compared by one-way analysis of variance. Results At 7 days after NMDA injection, the density of RGC in retinal patch of low-dose chloroquine group was significantly higher than that of intraperitoneal injection of PBS control group (F=54.41, P 0.05). The visual acuity was higher than control group, and the difference was statistically significant (F=9.10, P<0.05). The amplitude of PhNR-wave was significantly higher in low-dose chloroquine group than that of the control group (F=17.60, P<0.01). The mRNA level of inflammatory factor and GFAP positive signal was also significantly lower than that of the control group (F=23.66, P<0.05). The amplitude of PhNR-wave, the expression of GFAP (F=110.20, P<0.01) and the mRNA level of inflammatory factors (F=167.60, 17.78; P<0.01) in the high-dose chloroquine group were higher than the other two groups, and the differences were statistically significant. Conclusions In NMDA injury retinal model, low-dose chloroquine significantly increased the survival and physiological function of RGC, and the mechanism may be related to the inhibition of glial activation and inflammatory response. High-dose of chloroquine would aggravate the apoptosis of RGC. Key words: Chloroquine; Retinal ganglion cells; N-methylaspartate; Toxic actions; Animal experimentation

The protective effect and mechanism of rapamycin in the rat model of IgA nephropathy

Background: The pathogenesis of the development of IgA nephropathy has not been clear up to now. At present, some studies revealed that the mTOR pathway may participate in IgA nephropathy; however, the mechanism has not been systematically studied. In this study, we established an IgAN rat model to investigate the protective effects of rapamycin as a new type of immunosuppressant, as well as its therapeutic mechanisms.Methods: After the establishment of IgA nephropathy model, rats were treated with different concentrations of rapamycin, and the protective effect of different concentrations of rapamycin on renal function of the rats was observed. The deposition of IgA was observed by immunofluorescence. The kidney expression of Akt and p70S6k proteins in mTOR pathway was examined using the western blot assay after rapamycin treatment.Results: Morphology and immunofluorescence confirmed that the rat model of IgA nephropathy was successfully established. In particular, the level of proteinuria decreased with the increase of the dose of rapamycin, as well as the deposition of IgA in glomeruli. Moreover, the western blot analysis indicated that the expression of p70S6K in the downstream of mTOR pathway decreased and the upstream protein AKT of the mTOR pathway was overexpressed in the rats model.Conclusion: We found that rapamycin has protective effects in the IgA nephropathy rat model in a dose-dependent manner. In addition, the result of western blot assay suggested that rapamycin may display its therapeutic effects through interfering the AKT-mTOR-p70S6K signaling pathway.

Protective effect of different concentrations of normobaric oxygen in early cerebral ischemic-reper-fusion injury rat model

Objective To observe the protective effect of different concentrations of normobaric oxygen (NBO) on rats with acute cerebral ischemia reperfusion in early time.Methods 120 rats were divided randomly into high,medium and low NBO treatment groups and model group.Every group was divided into 3,6,12,24,48,and 72 h subgroups after ischemia reperfusion (n =5 each).The neurologic impairment scores were assessed in rats before and after oxygen inhalation,and the expression of Caspase-9 and Caspase-3 protein was detecte 4 d by using immunohistochemistry.Results Neurologic impairment scores in NBO treatment groups were decreased to varyoing degrees.There was statistically significant difference beween the high NBO treatment group (1.40 ±0.55,1.20 ±0.45,1.40 ±0.55,1.20 ±0.84,1.20 ±0.84,1.00±0.71) and the model group (2.20 ±0.45,2.20 ±0.84,2.20 ±0.45,2.20 ±0.45,2.80 ±0.84,2.60 ± 0.54) at each time point (P ＜ 0.05).The expression of Caspase-9 and Caspase-3 protein in the high,medium and low NBO treatment groups was lower than in the model group (P ＜ 0.05) at each time point.Conclusion The high concentration of NBO could reduce the expression of Caspase-9 and Caspase-3 protein obviously in rats with acute cerebral ischemia reperfusion and enhance the neurological function in early time,and medium and low concentrations of NBO had obvious effect delay as compared with high concentration of NBO. Key words: Normobaric oxygen; Cerebral ischemia; Reperfusion injury; Caspase-9 ; Caspase-3

Protective effect of Punica granatum L. (pomegranate) fruit extracts and ellagic acid against cytotoxicity induced by methyl tertiary butyl ether in isolated rat hepatocytes

Methyl tertiary butyl ether (MTBE) is the most widely used motor vehicle fuel oxygenate since it reduces automobile emissions of carbon monoxide and compounds involved in photochemical smog formation. Although recently banned in some developed countries, the increasing use of MTBE in Iran and many developing countries raised concern over its health safety. The protective effects of different concentrations (100, 150, 200 or 300 μg/ml) of aqueous and ethanolic extracts of Punica granatum L. fruit and also ellagic acid (120 μM) as one of its main components were examined against oxidative stress toxicity induced by MTBE in isolated rat hepatocytes. Addition of MTBE to isolated rat hepatocytes results in reactive oxygen species (ROS) formation, rapid glutathione (GSH) depletion, mitochondrial membrane potential collapse and lysosomal membrane rupture before hepatocyte lysis ensues. All Punica extracts and ellagic acid protected the hepatocyte against all cytotoxicity markers, including cell lysis, ROS generation, lipid peroxidation, GSH depletion, mitochondrial membrane potential decrease and lysosomal membrane oxidative damage. Punica aqueous extract (100, 150, 200 or 300 μg/ml) was more effective than its ethanolic extract (100, 150, 200 or 300 μg/ml) but similar to ellagic acid (120 μM) at protecting hepatocytes against MTBE-induced cytotoxicity, ROS generation, lipid peroxidation, GSH depletion, mitochondrial membrane potential reduction and lysosomal membrane damage. However, there were no significant differences between both Punica granatum L. fruit extracts and ellagic acid at hydrogen peroxide (H2O2) scavenging activity. These results suggest a hepatoprotective role for Punica extracts against liver injury resulting from MTBE-induced oxidative stress.

Protective Effect of Alpha-Tocopherol Isomer from Vitamin E against the H2O2Induced Toxicity on Dental Pulp Cells

The aim of this study was to evaluate the protective effects of different concentrations of vitamin E alpha-tocopherol (α-T) isomer against the toxicity of hydrogen peroxide (H2O2) on dental pulp cells. The cells (MDPC-23) were seeded in 96-well plates for 72 hours, followed by treatment with 1, 3, 5, or 10 mM α-T for 60 minutes. They were then exposed or not to H2O2 for 30 minutes. In positive and negative control groups, the cells were exposed to culture medium with or without H2O2 (0.018%), respectively. Cell viability was evaluated by MTT assay (Kruskal-Wallis and Mann-Whitney tests; α = 5%). Significant reduction of cell viability (58.5%) was observed in positive control compared with the negative control. Cells pretreated with α-T at 1, 3, 5, and 10 mM concentrations and exposed to H2O2 had their viability decreased by 43%, 32%, 25%, and 27.5%, respectively. These values were significantly lower than those observed in the positive control, thereby showing a protective effect of α-T against the H2O2 toxicity. Overall, the vitamin E α-T isomer protected the immortalized MDPC-23 pulp cells against the toxic effects of H2O2. The most effective cell protection was provided by 5 and 10 mM concentrations of α-T.

Nicotine-induced cessation of embryonic development is reversed by γ-tocotrienol in mice.

BackgroundThis study aimed to evaluate the adverse effects of various doses of nicotine and protective effects of different concentrations of γ-tocotrienol (γ-TCT) on in vitro embryonic development and lipid peroxidation in mice.Material/MethodsA) Effects of various doses of nicotine on in vitro embryonic development: Female mice were treated with 1.0, 3.0, or 5.0 mg/kg/day nicotine for 7 consecutive days. Animals were superovulated, cohabited overnight, and sacrificed. Embryos were cultured in vitro. Plasma was assayed. B) Effects of concomitant treatment of nicotine concurrently with various doses of γ-TCT on in vitro embryonic development: Female mice were treated with nicotine (5.0 mg/kg/day), gavaged γ-TCT of 30, 60, or 90 mg/kg/day or nicotine concurrently with γ-TCT of 3 different doses for 7 consecutive days. Animals were superovulated, cohabited overnight, and sacrificed. Embryos were cultured and plasma was assayed.ResultsA) Effects of various doses of nicotine on in vitro embryonic development: Number of hatched blastocysts decreased in 1.0 and 3.0 mg/kg/day nicotine groups. Nicotine at 5.0 mg/kg/day stopped embryo development at morula. MDA concentrations increased following all nicotine doses. B) Effects of concomitant treatment of nicotine concurrently with various doses of γ-TCT on in vitro embryonic development: Embryo development was completed in all groups. MDA concentration increased only in the group treated with nicotine concurrently with 30 mg/kg/day γ-TCT.ConclusionsNicotine impairs in vitro embryo development and increases MDA in plasma. The deleterious impact of nicotine on embryo development is reversed by supplementing γ-TCT concurrently with nicotine.

Protective Efficacy of Aqueous Extract of Naringi Crenulata Leaves against Shampoo (Endocrine Disruptor) Induced Toxicity in Labeo Rohita

The protective effect of different concentrations of leaf extracts of N. crenulata was tested against shampoo induced toxicity. 250mg/L/day of leaf extract brought the protein (25.7 ± 0.11), carbohydrate (30.7 ± 0.8) and lipid (5.13 ± 0.4) contents in its normal level in shampoo intoxicated fish. However, in shampoo intoxicated fish lipid content was found highest and decreasing with the increasing concentration of leaf extracts. Length and weight increased with the increase of leaf extract concentration and was observed equal to normal at 250 mg/L (length 6.63 ± 0.2, weight 3.7 ± 0.7). The results also showed that there is positive correlation between length and weight gain, leaf extract concentration and length gain. These results indicate that aqueous leaf extract of N. crenulata acts as an antidote against shampoo toxicity in L. rohita.

Protective effects and mechanism of hyperin on CoCl2-induced PC12 cells

To investigate the neuroprotective effects and mechanism of hyperin on CoCl2-induced hypoxic/ischemic PC12 cells. CoCl2 was used to treat rat pheochromocytoma cell line (PC12) to investigate the protective effects of different concentrations of hyperin. Hyperin could significantly inhibit CoCl2-induced cytotoxicity and apoptosis on PC12 cells in a dose-dependent manner, by inhibiting reactive oxygen species (ROS) production, increasing mitochondrial membrane potential (MMP), and inactivating caspase-3 and poly ADP-ribose polymerase (PARP). Hyperin could inhibit CoCl2-induced cytotoxicity and apoptosis on PC12 cells, show neuroprotective effects on hypoxic/ischemic neural injuries.

A Search for Hepatoprotective Activity of Fruit Extract of Mangifera indica L. Against Oxidative Stress Cytotoxicity

Mango (Mangifera indica L.) and their components are commonly used in folk medicine for many curative effects. The protective effects of different concentrations of aqueous extract of Mangifera indica L. fruit (Mango Extract) (20, 50 and 100 microg/ml) and also gallic acid (100 microM) as a pure compound in the extract were examined against oxidative stress toxicity induced by cumene hydroperoxide (CHP) in isolated rat hepatocytes. The extracts and gallic acid (100 microM) protected the hepatocyte against all oxidative stress markers including cell lysis, ROS generation, lipid peroxidation, glutathione depletion, mitochondrial membrane potential decrease, lysosomal membrane oxidative damage and cellular proteolysis. Mango Extracts (20, 50 and 100 microg/ml) were more effective than gallic acid (100 microM) in protecting hepatocytes against CHP induced lipid peroxidation. On the other hand gallic acid (100 microM) acted more effective than Mango Extracts (20, 50 and 100 microg/ml) at preventing lysosomal membrane damage. In addition H(2)O(2) scavenging effect of all extracts were determined in hepatocytes and compared with gallic acid (100 microM). There were no significance differences (P<0.05) between all plant extracts and gallic acid (100 microM) in H(2)O(2) scavenging activity. These results suggest a hepatoprotective role for Mango Extract against liver injury associated with oxidative stress.