Effect of ammonium pyrrolidine dithiocarbamate (PDTC) on NF-κB activation and CYP2E1 content of rats with immunological liver injury

Context: Ammonium pyrrolidine dithiocarbamate (PDTC) is a potent inhibitor of nuclear factor-κB (NF-κB). Recent studies have shown that NF-κB plays an essential role in the regulation of genes whose products are involved in the pathogenesis of immunological liver injury.

Objective: To study the function of NF-κB in immunological liver injury of rat model and its effect on CYP2E1 content and metabolic activity.

Materials and methods: The present study investigated the effect of passivating NF-κB activation on CYP2E1 using Bacillus calmette Guérin (BCG)-induced immunological liver injury in Sprague–Dawley rats measured in terms of enzyme levels. The degree of hepatic injury of rats was measured by using biochemical parameters, hepatic tissue pathological changes, and physiological parameters. Protein localization of liver NF-κB was detected by immunohistochemical assay. Western blot analysis was used to detect the protein expression of NF-κB, IκBα, iNOS, and CYP2E1. The content of CYP2E1 of homogenate in the rat liver was detected by ELISA assay and the enzyme kinetics of CYP2E1 probe drug chlorzoxazone was evaluated by high-performance liquid chromatography (HPLC) assay.

Results: The results showed that BCG-pretreatment (125?mg/kg) significantly (p?p?in vivo. Moreover, PDTC (ED₅₀: 76?mg/kg) dose dependently inhibited down-regulation of CYP2E1 (p?Conclusion: Passivation of NF-κB can inhibit the down-regulation of CYP2E1 and iNOS to induce in rat liver tissue with immunological liver injury; NF-κB may be involved in the CYP2E1 regulation through iNOS.     

Anti-inflammatory activity of Ternstroemia gymnanthera stem bark extracts in bacterial lipopolysaccharide-stimulated RAW264.7 murine macrophage cells

Context: Ternstroemia gymnanthera Sprague (Theaceae) possesses various known pharmacological properties. However, its anti-inflammatory activity has not been reported.

Objective: The anti-inflammatory activity of Ternstroemia gymnanthera stem bark aqueous extract (TGSBE) was evaluated using LPS-stimulated RAW264.7 macrophages.

Materials and methods: Cytotoxicity was assessed by MTT assay after 24?h with TGSBE (25–200?μg/mL). Further testing used TGSBE at 100 and 200?μg/mL. Griess and ELISA methods after 24?h with TGSBE determined NO and cytokine levels, respectively; then, mRNA levels (iNOS &; cytokines) were analyzed by Quantitative-PCR after 12?h. NF-κB and MAPK were assessed by immunoblotting after TGSBE treatment for 12?h, followed by LPS for 30?min. Immunofluorescence assay was also performed for NF-κB. ROS and MMP, after 12?h with TGSBE, were determined by flow cytometry. The antioxidant potential of TGSBE was analyzed by ABTS assay. The Folin–Ciocalteu method determined the total phenolic content of TGSBE. LPS concentration was 0.5?μg/mL.

Results: TGSBE at 200?μg/mL showed about 96.2% viability while suppressing the production of NO (88.99%), TNFα (24.38%), IL-6 (61.70%) and IL-1β (55.12%) and gene expression by 67.88, 45.24, 65.84, and 70.48%, respectively. TGSBE decreased ROS (79.26%) and improved MMP (48.01%); it inhibited translocation of NF-κB and MAPK activation. Radical scavenging activity was 50% at 402.17?μg/mL (ascorbic acid standard: 88.8?μg/mL). Total phenolic content was 240.9?mg GAE/g.

Discussion and conclusion: TGSBE suppresses the inflammatory response by inhibiting the NF-κB and MAPK cascades exhibiting therapeutic potential to treat inflammatory diseases associated with increased activation of macrophages.     

Implication of thymoquinone as a remedy for polycystic ovary in rat

Context: Thymoquinone (TQ), an active component of Nigella sativa L. (Ranunculaceae), possesses anti-inflammatory and anti-oxidative properties. Polycystic ovary syndrome exhibits chronic inflammatory behavior, thus might involve nuclear factor kappa B (NF-κB) signaling and related molecular factors.

Objective: The objective of the present study is to investigate and validate the effect of TQ in polycystic ovary (PCO) rat.

Materials and methods: To validate the effect of TQ (1?µM/ml), NF-κB activation, COX2 (cyclooxygenase-2) expression and reactive oxygen species (ROS) induction were studied in the KK1 cell line. To evaluate the effect of TQ (2?mg/200?µl olive oil/rat; sc) with an in vivo system, ovulation rate, levels of key ovulation mediators, and ovarian gelatinases activity were compared in superovulated, PCO, and RU486?+?TQ-treated Wistar rats.

Results: In vitro studies showed that NF-κB nuclear translocation, COX2, and ROS expression were repressed via TQ supplementation in RU486-treated KK1 cells. Pretreatment of TQ in the PCO rat model induced significant restoration of normal physio-molecular behavior of ovary, such as reduced cysts formation, increased ovulation rate, and normalization of key ovarian factors [like TNF-α-stimulated gene/protein 6, hyaluronan, hyaluronan-binding protein 1, COX2, matrix metalloproteinases (membrane type 1-matrix metalloproteinase, MMP9 and MMP2)], tissue inhibitor of metalloproteinases (TIMP-1 and TIMP-2), and gelatinases (like MMP9 and -2) activity during follicular maturation.

Discussion and conclusion: Overall, most of the above molecular changes are regulated via NF-κB pathway, thus TQ, due to its modulatory effect on the NF-κB signaling, could elevate normal ovarian phenotype and physiological function in the PCO model, indicating its remarkable potential as a remedy for rat PCO.     

Altered oxidative status and integrin expression in cyclosporine A-treated oral epithelial cells

Abstract

Background and objective: Cyclosporine A (CsA) is an immunosuppressive agent administered to transplant patients. A well-known reported oral side effect of CsA consumption is gingival overgrowth (GO). Changes in the expression of integrins occurring in the gingiva following CsA treatment have been reported but these reports are mainly concerned with the connective tissue of the gingiva. In this study we targeted the alterations in the oral epithelium using KB cells, an oral epithelial cell line.

Methods: Cultured oral epithelial cells were treated with increasing concentrations of CsA (0.1, 1 and 10?µg/mL) and the molecular changes involving antioxidant enzymes [glutathione peroxidase (GPx) and glutathione reductase (GR)] and the level of reactive oxygen species (ROS) were measured. Quantitative real-time PCR was used to assess the expression of selected integrins (α₂, α₅ and β₁).

Results: At CsA concentration above 0.1?µg/mL GPx demonstrated an increase in activity while GR activity and the level of reduced glutathione were diminished (p?<?0.05). α₅ and β₁ integrin were downregulated at all treatment concentrations of CsA while α₂ integrin presented this effect at concentrations above 1?µg/mL (p?<?0.05).

Conclusion: The results suggest a possible role for oxidative stress and the altered expression of integrins in the pathology of CsA-induced gingival overgrowth.     

Effects of erdosteine on cyclosporine-A-induced hepatotoxicity in rats

Cyclosporine A (CsA) is a potent immunosuppressive agent used for organ transplantations and various autoimmune disorders. However, hepatotoxicity due to CsA remains one of the major side effects. The use of antioxidants reduces the adverse effects of CsA. The aim of this study was to determine the protective effects of erdosteine on CsA-induced liver injury through tissue oxidant/antioxidant parameters and to evaluate light microscopic alterations in rat-liver tissues. Rats were randomly divided into four experimental groups: The control group received sunflower oil (2?mL/kg/day, per orally; p.o.), while the other groups were treated with CsA (25?mg/kg/day, p.o.) or erdosteine (10?mg/kg/day, p.o.) or CsA+erdosteine, respectively. Serum aspartate aminotransferase and alanine aminotransferase levels, tissue malondialdehyde and nitric oxide levels, and superoxide dismutase, glutathione peroxidase and catalase enzyme activities were measured. Histological examination was performed. CsA caused a significant deterioration in the hepatic function tests, morphology, and gave rise to severe oxidative stress in the liver. Erdostein significantly improved the functional and histological parameters and attenuated the oxidative stresss induced by CsA. Erdostein protects liver tissue against oxygen free radicals and prevents hepatic dysfunction and morphological abnormalities associated with chronic CsA administration.     

Cannabinoid receptor agonist WIN55,212-2 and fatty acid amide hydrolase inhibitor URB597 ameliorate neuroinflammatory responses in chronic cerebral hypoperfusion model by blocking NF-κB pathways

The present study explored the protective effects of cannabinoid receptor agonist WIN55,212-2 (WIN) and fatty acid amide hydrolase inhibitor URB597 (URB) against neuroinflammation in rats with chronic cerebral hypoperfusion (CCH). Activated microglia, astrocytes, and nuclear factor kappa B (NF-κB) p65-positive cells were measured by immunofluorescence. Reactive oxygen species (ROS) was assessed by dihydroethidium staining. The protein levels of cluster of differentiation molecule 11b (OX-42), glial fibrillary acidic protein (GFAP), NF-κB p65, inhibitor of kappa B alpha (IκB-a), IκB kinase a/β (IKK a/β), phosphorylated IKK a/β (p-IKK a/β), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-α, and interleukin-1β (IL-1β) were examined by western blotting or enzyme-linked immunosorbent assay. All the protein levels of OX-42, GFAP, TNF-a, IL-1β, COX-2, and iNOS are increased in CCH rats. WIN and URB downregulated the levels of OX-42, GFAP, TNF-α, IL-1β, COX-2 and iNOS and inhibited CCH-induced ROS accumulation in CCH rats, indicating that WIN and URB might exert their neuroprotective effects by inhibiting the neuroinflammatory response. In addition, the NF-κB signaling pathway was activated by CCH in frontal cortex and hippocampus, while the aforementioned changes were reversed by WIN and URB treatment. These findings suggest that WIN and URB treatment ameliorated CCH-induced neuroinflammation through inhibition of the classical pathway of NF-κB activation, resulting in mitigation of chronic ischemic injury.

     

Angiotensin-converting enzyme-2 overexpression attenuates inflammation in rat model of chronic obstructive pulmonary disease

Abstract

Objective: To investigate the anti-inflammatory effects of angiotensin-converting enzyme 2 (ACE2) overexpression on rat model of chronic obstructive pulmonary disease (COPD), and explore underlying mechanism.

Methods: The rat COPD model was established by cigarette smoking using a total body exposure method. A total of 64 male Wistar rats were randomly divided into four groups: normal, COPD, Ad-ACE2 and Ad-EGFP groups. The COPD model rats (including COPD, Ad-ACE2 and Ad-EGFP groups) received an intratracheal injection of normal saline, Ad-ACE2 and Ad-EGFP, respectively. The normal group underwent the same procedure but received an intratracheal injection of normal saline only. Pulmonary function tests, lung histopathology analysis, malondialdehyde (MDA) and reactive oxygen species (ROS) level, ACE2 mRNA and protein expression level, inflammatory cytokines and related signaling pathway proteins were measured.

Results: COPD rats showed impairment of lung function as evidenced by decreased ratio of forced expiratory volume at 0.3?s and forced vital capacity (FEV_0.3/FVC) and dynamic lung compliance (Cldyn), increased resistance inspiration (Ri) and resistance expiration (Re) as compared with the normal group, accompanying with reduced ACE2 mRNA expression, elevated ROS and MDA, elevated inflammatory cytokines levels (tumor necrosis factor α, TNF-α; interleukin-8, IL-8; IL-2 and IL-1β) and activation of nuclear factor-κB (NF-κB) and p38 MAPK (mitogen activated protein kinases) pathway in lung tissues. ACE2 overexpression through Ad-ACE2 infusion significantly attenuated the inflammatory response in lung tissues of COPD model rats.

Conclusion: ACE2 could attenuate COPD inflammatory process induced by cigarette smoke through reduction of oxidative stress and inhibition of NF-κB and p38 MAPK pathway activation.     

Synergistic protective effect of folic acid and vitamin B12 against nicotine-induced oxidative stress and apoptosis in pancreatic islets of the rat

Context: Nicotine is an abundant and most significant component of cigarette smoke. Epidemiological evidence strongly suggests an association between cigarette smoking and pancreatic injury, although effects of smoking on endocrine pancreas are still controversial.

Objective: We examined the impact and underlying mechanisms of action of folic acid and vitamin B₁₂ on nicotine-induced damage in pancreatic islets of rats.

Materials and methods: Male Wistar rats were treated with nicotine (3?mg/kg body weight/d, intraperitonealy) with or without folic acid (36?µg/kg body weight/d, orally) and vitamin B₁₂ (0.63?µg/kg body weight/d, orally) for 21?d. Fasting blood glucose, oral glucose tolerance test, HBA_1c, insulin, oxidative stress parameters, proinflammatory cytokines, and CRP level were measured. Histological evaluation, TUNEL assay, and immunohistochemical staining of NF-κB and caspase-3 were also performed.

Results: Folic acid and vitamin B₁₂ blunted the nicotine-induced impairment in fasting blood glucose (51–56% recovery), HbA_1c (64–76% recovery), oral glucose tolerance, insulin level (23–40% recovery), and islet cell counts (26–74% recovery) in rats. Moreover, folic acid in combination with vitamin B₁₂ also attenuated the nicotine-induced changes in markers of oxidative stress (17–88% recovery), TNF-α (40–99% recovery), and IL-6 level (47–65% recovery), CRP level (59–73% recovery), expression of NF-κB and caspase-3, and apoptosis in pancreatic islet cells.

Discussion and conclusion: The present study shows that folic acid and vitamin B₁₂ supplementation can reduce nicotine-induced impairment in glucose homeostasis and apoptosis and damage of pancreatic islet cells by modulating oxidative stress, levels of proinflammatory cytokines, and expression of NF-κB.     

钝化NF-κB的活化对酒精性肝损伤大鼠CYP3A的影响

目的研究核转录因子-κB(nuclear fastor kappa B,NF-κB)在酒精性肝损伤大鼠模型中的作用及对细胞色素P4503A(cytochrome P450 3A,CYP3A)代谢活力的影响。方法采用白酒灌胃法复制大鼠酒精性肝损伤模型,肝脏组织HE染色和血清中ALT和AST水平测定观测大鼠肝损伤情况,采用免疫组化法检测肝脏NF-κB的蛋白表达,通过HPLC法检测CYP3A的探针药物咪哒唑仑的血浆药物浓度,采用热板法观察大鼠舔足反应来体现咪哒唑仑的代谢情况,从而反映咪哒唑仑特异性代谢酶CYP3A的代谢活力。结果酒精性肝损伤模型组表现为轻度脂肪变性和炎症坏死,NF-κB明显活化,CYP3A代谢活力增强(P<0.05);钝化NF-κB的活化后,肝脏脂肪变性程度明显减轻,CYP3A代谢活力下调(P<0.05)。结论钝化NF-κB活化,可以减轻酒精性肝损伤的程度,并下调酒精性肝损伤导致的CYP3A代谢活性的增强。     

Protection by genistein on cortical neurons against oxidative stress injury via inhibition of NF-kappaB,JNK and ERK signaling pathway

Abstract

Context: Genistein, one of the isoflavones derived from soybean seeds, has been reported to exert multiple bioactivities. However, the mechanism of its action on the central nervous system is not fully understood.

Objective: To investigate the cytoprotection of genistein and its molecular mechanism against H₂O₂-induced cell death in primary rat cortical neurons.

Materials and methods: Genistein (0.01, 0.1, and 1?μM) were added into the primary rat neurons 24?h before and co-cultured with 500?μM H₂O₂ for 1?h. Neuronal injury was assessed by MTT, lactate dehydrogenase (LDH) assay, and Hoechst33258 staining. Intracellular reactive oxygen species (ROS) generation induced by H₂O₂ was determined. Neuronal apoptosis was evaluated by Bcl-2/Bax ratio as well as by caspase-9 and caspase-3 activities. The protein levels and phosphorylation of NF-κB/p65, IκB, JNK, and ERK were detected by western blots.

Results: Genistein pretreatment attenuated H₂O₂-mediated neuronal viability loss, nuclear condensation, and ROS generation in a concentration-dependent manner. Genistein exerted anti-apoptotic effects by reversing the apoptotic factors Bcl-2 and Bax ratio, along with the suppression of caspase-9 and caspase-3 activities. In addition, genistein down-regulated the expression of NF-κB/p65, and suppressed the phosphorylation of p65 and IκB. Genistein also inhibited H₂O₂-induced activation of the MAPK-signaling pathway including JNK and ERK.

Discussion and conclusion: The results indicated that genistein effectively protects cortical neurons against oxidative stress at least partly via inactivation of NF-κB as well as MAPK-signaling pathways, and suggested the possibility of this antioxidant for the prevention and treatment of stroke.     

Therapeutic effect of liposomal-N-acetylcysteine against acetaminophen-induced hepatotoxicity

Abstract

Background: Acetaminophen (APAP) is an antipyretic analgesic drug that when taken in overdose causes depletion of glutathione (GSH) and hepatotoxicity. N-acetylcysteine (NAC) is the antidote of choice for the treatment of APAP toxicity; however, due to its short-half-life repeated dosing of NAC is required.

Purpose: To determine whether a NAC-loaded liposomal formulation (Lipo-NAC) is more effective than the conventional NAC in protecting against acute APAP-induced hepatotoxicity.

Methods: Male Sprague–Dawley rats were challenged with an intragastric dose of APAP (850?mg/kg b.wt.); 4?h later, animals were administered saline, NAC, Lipo-NAC or empty liposomes and sacrificed 24?h post-APAP treatment.

Results: APAP administration resulted in hepatic injury as evidenced by increases in plasma bilirubin, alanine (AST) and aspartate (ALT) aminotransferase levels and tissue levels of lipid peroxidation and myeloperoxidase as well as decreases in hepatic levels of reduced GSH, GSH peroxidase and GSH reductase. Treatment of animals with Lipo-NAC was significantly more effective than free NAC in reducing APAP-induced hepatotoxicity. Histological evaluation showed that APAP caused periacinar hepatocellular apoptosis and/or necrosis of hepatocytes around the terminal hepatic venules which was reduced by NAC treatment, the degree of reduction being greater for Lipo-NAC.

Conclusion: These data suggest that administration of Lipo-NAC ameliorated the APAP-induced hepatotoxicity.     

Effect of iNOS and NF-κB gene silencing on β-cell survival and function

Purpose: Type I diabetes results from β-cell death and dysfunction, induced by infiltration of immune cells and local production of inflammatory cytokines. Therefore, we investigated the effect of iNOS and NF-κB gene silencing on β-cell survival and function.

Methods: Rat insulinoma INS-1E cells were transfected with chemically synthesized siRNA after complex formation with Lipofectamine 2000. Cells were then treated with a cocktail of inflammatory cytokines (IL-1β+ TNF-α+ IFN-α), and glucose stimulated-insulin response and viability were determined. iNOS and NF-κB gene expression was assessed at mRNA level by real time RT-PCR. The effect of gene silencing was also correlated with cytokine-induced NO production and apoptosis.

Results: Transfection of INS-1E cells with siRNAs silenced iNOS and NF-κB gene expression and reduced NO production in a sequence-specific manner without causing significant loss of cell viability and function. However, the abrogation of NO production did not prevent INS-1E cells from cytokine-induced apoptosis, suggesting that this event may not be totally dependent on NO production.

Conclusion: The gene silencing approach presented here is capable of attenuating the effects of inflammatory cytokines, such as iNOS expression and NO production and it will help to identify new target genes to improve islet transplantation.     

Anti-inflammatory effects of Cynanchum taiwanianum rhizome aqueous extract in IL-1β-induced NRK-52E cells

Context: Cynanchum taiwanianum T. Yamaza (Asclepiadaceae) is a medicinal herb used in folk medicine for the treatment of several inflammation-related diseases such as hepatitis and dermatitis in Taiwan.

Objective: In the present study, we investigated the anti-inflammatory effect of C. taiwanianum T. Yamaza rhizome aqueous extract (CTAE).

Materials and methods: The present study investigated the anti-inflammatory effect of CTAE using IL-1β-induced NRK-52E cells. Production of NO and PGE₂ by ELISA, the mRNA and protein expression of iNOS and COX-2, phosphorylation of IκBα, and activation of NF-κB by RT-PCR and western blotting were determined.

Results: The CTAE significantly (P?<?0.05) inhibited NO and PGE₂ production (decreased by 46.1% and 51%, respectively), and also significantly (P?<?0.05) attenuated protein and mRNA expression of iNOS and COX-2 (decreased by 90% and 55% for iNOS and by 72% and 74%% for COX-2, respectively) in IL-1β-induced NRK-52E cells, in a dose-dependent manner, without obvious cytotoxic effects. Furthermore, the CTAE suppressed the NF-κB nuclear translocation, in terms of inhibition of IκBα phosphorylation.

Discussion and conclusion: Our results provided evidence for its folkloric uses and suggest that the anti-inflammatory activities of CTAE may result from the inhibition of inflammatory mediators, such as NO and PGE₂, and an upstream suppression of a NF-κB-dependent mechanism, might be involved.     

Macrophage activation by polysaccharides from Atractylodes macrocephala Koidz through the nuclear factor-κB pathway

Abstract

Context: Atractylodes macrocephala Koidz is a traditional herb. Atractylodes macrocephalaon polysaccharides (AMP) have been found to enhance immunity and improve heart function. However, the mechanisms of the immunomodulatory effect have not been investigated.

Objective: We examined whether AMP activated macrophages and explored the mechanisms of activation.

Materials and methods: AMP was prepared and evaluated its immunomodulatory activity (25, 50, 100, and 200?μg/mL) by detecting the phagocytosis and the production of tumor necrosis factor-α (TNF-α), IFN-γ, and nitric oxide (NO) in RAW264.7 macrophages. Furthermore, the role of nuclear factor-κB (NF-κB) pathway was examined in regulating TNF-α and NO production.

Results: The phagocytosis of macrophages was enhanced by AMP in a dose-dependent manner and the maximal phagocytosis of macrophages occurred at concentrations of 100 and 200?μg/mL. NO, TNF-α, and IFN-γ release was also found to be dose dependent by increasing concentrations of AMP and reached the peak at a concentration of 200?μg/mL. In addition, AMP induced inhibitor kappaB (IκB) degradation and the activation of NF-κB by p65 nuclear translocation, and then the activation of NF-κB in nucleus peaked at a concentration of 200?μg/mL. Besides, NF-κB-specific inhibitor pyrrolidine dithiocarbamate (PDTC) decreased AMP-induced NO and TNF-α production.

Discussion and conclusion: These data suggest that AMP may modulate macrophage activities by stimulating NF-κB or activating NF-κB-dependent mechanisms.     

The protective effect of erdosteine against cyclosporine A-induced cardiotoxicity in rats

Cyclosporine A (CsA) is a frequently used immunosuppressive agent in transplant medicine to prevent rejection and in the treatment of autoimmune diseases. However, CsA generates reactive oxygen species, which causes nephrotoxicity, hepatotoxicity and cardiotoxicity. The use of antioxidants reduces the adverse effects of CsA. The aim of this study is to determine the protective effects of erdosteine on CsA-induced heart injury through tissue oxidant/antioxidant parameters and light microscopic evaluation in rats. CsA cardiotoxicity was induced by administrating an oral dose of 15mg/kg CsA daily for 21 days. The rats were divided into four groups: control group (n=4), CsA administrated group (15mg/kg, n=5), CsA+erdosteine administrated group (10mg/kg day orally erdosteine, n=4) and only erdosteine administrated group (10mg/kg day orally n=5). CsA treated rats showed increase in the number of infiltrated cells and disorganization of myocardial fibers with interstitial fibrosis. The number of infiltrated cells, disorganization of myocardial fibers and interstitial fibrosis was diminished in the hearts of CsA-treated rats given erdosteine. The malondialdehyde, the protein carbonyl content and nitric oxide levels were increased in the cyclosporine A group in comparison with the control and CsA plus erdosteine groups. The activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were higher in CsA plus erdosteine group than CsA group. However, the CAT, GSH-Px and SOD activities were significantly lower in CsA group than in control group and erdosteine group. These results suggest that erdosteine has protective effect against CsA-induced cardiotoxicity.     

Hepatoprotective effect of Stachys pilifera ethanol extract in carbon tetrachloride-induce hepatotoxicity in rats

Context: Stachys pilifera Benth (Lamiaceae) has long been used to treat infectious diseases, respiratory and rheumatoid disorders in Iranian folk medicine. Antitumor and antioxidant activity of the plant have been reported.

Objective: The study was designed to assess the hepatoprotective activity of ethanol extract of Stachys pilifera in carbon tetrachloride (CCl₄)-induced hepatotoxicity in rats.

Materials and methods: The rats were randomly divided into six equal groups (n?=?7). Group I was treated with normal saline; Group II received CCl₄ (1?mL/kg. i.p., twice a week) for 60 consecutive days; Groups III, IV and V were given CCl₄ plus Stachys pilifera (100, 200 and 400?mg/kg/d,p.o.); Group VI received the extract (400?mg/kg/d, p.o.). Histopathological analysis and measurement of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), malondialdehyde (MDA), total protein (TP) and albumin (ALB) were performed.

Results: CCl₄ caused a significant increase in the serum levels of AST, ALT, ALP and MDA as well as decreased ALB, and TP serum levels (p?4-elevated levels of ALT, AST, ALP and MDA (p?4 group (p4.

Discussion: The results revealed that the Stachys pilifera extract could provide considerable protection against CCl₄ hepatotoxicity in rats that may be related to its antioxidant properties.     

Effects of linalool on inflammation,matrix accumulation and podocyte loss in kidney of streptozotocin-induced diabetic rats

Abstract

Context: This study aimed to evaluate the renoprotective action of linalool (LIN) on streptozotocin (STZ)-induced diabetic rats.

Objective: The pathological changes in diabetic nephropathy (DN) include oxidative stress, renal injury, matrix accumulation and podocyte abnormalities. We investigated the renoprotective actions of LIN, a monoterpene alcohol, present in herbal essential oils in STZ-induced diabetic rats with renal injury.

Materials and methods: STZ-diabetic rats were administered LIN (25?mg/kg) for 45 days, after which the activities of key enzymes of glucose metabolism, collagen content, oxidative damage and expression of glucose transporter-1 (GLUT-1), transforming growth factor-β₁ (TGF-β₁), nuclear factor-κBp65 (NF-κB p65) and nephrin were analyzed.

Results: Diabetic rats displayed altered glucose metabolism, collagen accumulation and increased TGF-β₁ and NF-κB expression in kidney. LIN treatment restored glucose-metabolizing enzymes, collagen content and GLUT-1 expression and also prevented nephrin loss. LIN also rescued kidney from oxidative stress and inflammation by decreasing the expression of TGF-β₁ and NF-κB. Ultrastructural changes such as basement membrane thickening, reduction in podocyte number and loss of filtration barrier integrity in diabetic rats were mitigated by LIN.

Discussion and conclusion: The results of this study suggest that LIN can attenuate nephropathic changes induced in kidney of diabetic rats. These findings highlight the utility of LIN as a potential drug to treat renal damage in diabetic subjects.     

Puerarin mediates hepatoprotection against CCl4-induced hepatic fibrosis rats through attenuation of inflammation response and amelioration of metabolic function

This study was designed to evaluate the potential effects of puerarin (PR), an effective isoflavonoid compound purified from Pueraria lobata, in treating hepatic fibrosis (HF) rats induced by carbon tetrachloride (CCl₄, 2 mL kg⁻¹ d⁻¹). Compared to model control, PR treatment effectively lowered the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (Alb), total protein (TP) in HF rats. Masson stained analysis showed that the condition of HF rats was mitigated. Meanwhile, the tumor necrosis factor alpha (TNF-α), nuclear factor-kappa B (NF-κB) expressions were significantly down-regulated at protein level by PR intervention. Additionally, the activity of superoxide dismutase (SOD) was elevated, while the content of malondialdehyde (MDA) was lessened in liver tissue. As revealed by immunohistochemistry assay, PR therapy resulted in reduced production of transforming growth factor-βl (TGF-βl). Moreover, it also was attributed to decreased mRNA level of inducible nitric oxide synthase (iNOS) using RT-PCR analysis. These findings demonstrate that puerarin successfully reverses hepatotoxicity in CCl₄-induced HF rats via the underlying mechanisms of regulating serum enzymes and attenuating TNF-α/NF-κB pathway for anti-inflammation response, as well as improving metabolic function in liver tissue.