Protective and Anti-Aging Effects of 5 Cosmeceutical Peptide Mixtures on Hydrogen Peroxide-Induced Premature Senescence in Human Skin Fibroblasts.

Skin aging usually leads to the excessive deterioration of the dermal extracellular matrix, loss of antimicrobial function, loss of skin barrier function, and a series of inflammatory processes. Bioactive peptides have been widely used in cosmetics due to their protective effects on skin and efficient absorption. Combination of different peptides may lead to synergistic or antagonistic effects, so different formulas need to be designed and tested properly. In this study, 5 functional cosmeceutical peptides were tested on their individual and mixed activities to detect a suitable anti-aging and protective formula from our experiments. After the individual activity test, the optimal concentration is 200 µg/mL of carnosine for the superoxide dismutase (SOD) activity, 200 µg/mL of GHK peptide for the hydroxyproline (HYP) content activity, 100 µg/mL of acetyl tetrapeptide-5 for the angiotensin-converting enzyme 1 activity, 400 µg/mL of hexapeptide-11 for the HYP content activity, and 400 µg/mL of acetyl hexapeptide-3 for the catecholamine content activity. According to the optimal concentration of these 5 cosmeceutical peptides, 6 formulations of peptide mixtures were designed and tested for their anti-aging activities and protective effects against hydrogen peroxide-induced premature senescence in human skin fibroblasts. One of the cosmeceutical peptide mixtures (carnosine + acetyl tetrapeptide-5 + hexapeptide-11 + acetyl hexapeptide-3) significantly reduced the intracellular malondialdehyde and hydroxyl free radical contents and increased the HYP and human elastin contents as well as the enzymatic activities of SOD and glutathione peroxidase. Our study suggests that this formula of cosmeceutical peptide mixtures could be a promising agent for use in anti-aging and protective cosmetics.

Extraction and Identification of Two Flavonoids in Phlomoides hyoscyamoides as an Endemic Plant of Iran: The Role of Quercetin in the Activation of the Glutathione Peroxidase, the Improvement of the Hydroxyproline and Protein Oxidation in Bile Duct-Ligated Rats.

BACKGROUND: Cholestatic liver disease, a serious chronic condition that develops progressive hepatic degeneration through free radicals. OBJECTIVE: The present study was designed to extract and identify two flavonoids in Phlomoides hyoscyamoides plant, native to Iran and evaluate the role of quercetin identified on the liver injury among bile ductligated rats. METHODS: This study was conducted on 25 male Wistar rats within three groups of sham control, mere bile duct-ligated, and bile duct-ligated with quercetin. The bile duct-ligated animals received quercetin at a dose of 50 mg/kg/day for 10 days, followed by biochemical tests, oxidative stress markers, activity of antioxidant enzymes and hematoxylin and eosin staining. Molecular docking was used to explore the interactive behavior of quercetin with glutathione peroxidase. RESULTS: According to analyses of the obtained extract, two main active ingredients of P. hyoscyamoides were rutin and quercetin. Bile duct-ligated group showed a significant liver necrosis, a clear increase in plasma and tissue oxidative stress parameters, and a decrease in glutathione peroxidase activity as compared to sham control group. Quercetin injection in bile duct-ligated rats resulted in significant decrease in hydroxyproline, protein carbonyl and histopathologic indexes and significant increase in glutathione peroxidase activity (P-value≤0.05). Based on the molecular docking, the quercetin was able to regulate the glutathione peroxidase activity. CONCLUSION: The quercetin acts as an enzyme inducer by renewing the glutathione peroxidase activity and inhibiting the oxidation of proteins and hence decreases the oxidative stress. These results could be a sign of confirming the positive role of quercetin in attenuating the liver damage and degeneration.

A Chinese Traditional Therapy for Bleomycin-Induced Pulmonary Fibrosis in Mice.

Pulmonary fibrosis is a chronic and fatal disease of lung tissue with high incidence and mortality in the world. The exploration of effective treatment for pulmonary fibrosis remains an urgent challenge. In our study, Qingfei Xieding was investigated as a novel Chinese traditional patent medicine against pulmonary fibrosis. A pulmonary fibrosis mouse model was constructed by injecting with bleomycin sulfate. Following Qingfei Xieding administration, lung samples were collected to assess pulmonary phenotype changes by analyzing lung coefficient, wet/dry, and histopathologic section. Levels of nitric oxide (NO), hydroxyproline (HYP), malondialdehyde (MDA), and total antioxidant capacity were measured to evaluate the degree of oxidation. A single-cell gel electrophoresis (SCGE) assay was used to evaluate bleomycin-induced DNA damage. Western blotting and real-time quantitative PCR were performed to determine the abundance of inducible nitric oxide synthase (iNOS), connective tissue growth factor (CTGF), alpha smooth muscle actin (α-SMA), and fibronectin (FN). In the present study, Qingfei Xieding administration significantly attenuated bleomycin-induced pulmonary fibrosis in mice by reducing lung coefficient, wet/dry, NO, HYP, and MDA as well as the expression of iNOS, CTGF, α-SMA, FN, and DNA damage. The results indicated that Qingfei Xieding is effective to resist oxidative damage and histopathologic lesion, serving a protection role on bleomycin-induced pulmonary fibrosis.

Appraisal on the wound healing activity of different extracts obtained from Aegle marmelos and Mucuna pruriens by in vivo experimental models.

AIM: The use of a simple and reproducible model is inevitable for an objective statement of the effects of external factors on wound healing. Hence, the present study was conducted to evaluate wound healing activities of sequential different extracts of Aegle marmelos leaves (AM) and Mucuna pruriens seeds (MP) by in vivo experimental models. MATERIALS AND METHODS: Wistar albino rats were subjected to excision, incision and dead space wounds measuring approximately 250 mm2, 3 cm and implanting sterilized polyvinyl chloride tube on the back of each rat near either side of the vertebral column respectively. The experimental animals were randomized into eight groups (n = 6), control, standard and treatment groups. Hydrogel of different extracts were applied topically once daily. The parameters observed were percentage of wound contraction, epithelization period, tensile strength, hydroxyproline content of the granulation tissue, and histological changes during wound healing. RESULTS: The statistical study revealed that in excision, incision, and dead space wound models all formulations have significant (P < 0.01) wound healing potential. However, methanolic extract formulation was found to be superior to all other treatments as evidenced by rapid wound contraction, lesser number of days required for complete epithelization, increased tensile strength and significant increase in hydroxyproline content. CONCLUSIONS: As compared to the reference standard treated group the wound healing process of the experimental groups was decelerated. All extracts obtained from AM and MP facilitated the wound healing process in all experimental models.

Combination of sorafenib and gadolinium chloride (GdCl3) attenuates dimethylnitrosamine(DMN)-induced liver fibrosis in rats.

BACKGROUND/AIMS: Liver sinusoidal endothelial cells (SECs), hepatic stellate cells (HSCs) and Kupffer cells (KCs) are involved in the development of liver fibrosis and represent a potential therapeutic target. The therapeutic effects on liver fibrosis of sorafenib, a multiple tyrosine kinase inhibitor, and gadolinium chloride (GdCl3), which depletes KCs, were evaluated in rats. METHODS: Liver fibrosis was induced in rats with dimethylnitrosamine, and the effects of sorafenib and/or GdCl3 in these rats were monitored. Interactions among ECs, HSCs and KCs were assessed by laser confocal microscopy. RESULTS: The combination of sorafenib and GdCl3, but not each agent alone, attenuated liver fibrosis and significantly reduced liver function and hydroxyproline (Hyp). Sorafenib significantly inhibited the expression of angiogenesis-associated cell markers and cytokines, including CD31, von Willebrand factor (vWF), and vascular endothelial growth factor, whereas GdCl3 suppressed macrophage-related cell markers and cytokines, including CD68, tumor necrosis factor-α, interleukin-1ß, and CCL2. Laser confocal microscopy showed that sorafenib inhibited vWF expression and GdCl3 reduced CD68 staining. Sorafenib plus GdCl3 suppressed the interactions of HSCs, ECs and KCs. CONCLUSION: Sorafenib plus GdCl3 can suppress collagen accumulation, suggesting that this combination may be a potential therapeutic strategy in the treatment of liver fibrosis.

Effect of SMAD7 gene overexpression on TGF-ß1-induced profibrotic responses in fibroblasts derived from Peyronie's plaque.

Transforming growth factor-ß1 (TGF-ß1) has been identified as one of the most important fibrogenic cytokines associated with Peyronie's disease (PD). The mothers against decapentaplegic homolog 7 (SMAD7) is an inhibitory Smad protein that blocks TGF-ß signaling pathway. The aim of this study was to examine the anti-fibrotic effect of the SMAD7 gene in primary fibroblasts derived from human PD plaques. PD fibroblasts were pretreated with the SMAD7 gene and then stimulated with TGF-ß1. Treated fibroblasts were used for Western blotting, fluorescent immunocytochemistry, hydroxyproline determination, and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assays. Overexpression of the SMAD7 gene inhibited TGF-ß1-induced phosphorylation and nuclear translocation of SMAD2 and SMAD3, transdifferentiation of fibroblasts into myofibroblasts, and quashed TGF-ß1-induced production of extracellular matrix protein and hydroxyproline. Overexpression of the SMAD7 gene decreased the expression of cyclin D1 (a positive cell cycle regulator) and induced the expression of poly (ADP-ribose) polymerase 1, which is known to terminate Smad-mediated transcription, in PD fibroblasts. These findings suggest that the blocking of the TGF-ß pathway by use of SMAD7 may be a promising therapeutic strategy for the treatment of PD.

The inhibitory effect of proanthocyanidin on soluble and collagen-bound proteases.

OBJECTIVE: This study evaluated the inhibitory effect of proanthocyanidin (PA), a natural collagen cross-linker, on soluble and matrix-bound proteases, which are responsible for progressive degradation of exposed collagen fibrils within the hybrid layer and resin-dentine bond failure over time. METHODS: The inhibitory effects of PA (1%, 2%, 3%, 4.5% and 6%) on soluble recombinant matrix metalloproteinases (MMP-2, -8 and -9) and cysteine cathepsins (cathepsin B and K) were evaluated using MMP and cysteine cathepsins fluorometric assay kits. Chlorhexidine (CHX) was used as an inhibitor control. The effect of PA on endogenous matrix-bound proteases was examined by determining the change in dry mass of demineralized dentine beams and solubilized collagen peptides over 30 days. Two-way ANOVA and Tukey multiple comparison tests were used to analyze the effect of PA and proteases on the percentage inhibition of soluble proteases (α=0.05). Kruskal-Wallis one-way ANOVA and Dunn's multiple comparison tests were used to analyse the effect of PA on loss of dry mass and hydroxyproline content over time (α=0.05). RESULTS: Proanthocyanidin inactivated more than 90% of soluble recombinant MMP-2, -8 and -9 and around 75-90% of cysteine cathepsin B and K, which was significantly higher than CHX (P<0.05). The inhibition of endogenous proteases by PA increased in a dose-dependent manner. The loss of dry mass and hydroxyproline release in the medium over time was the lowest in dentine beams pretreated with PA

Effects of zoledronate on irradiated bone in vivo: analysis of the collagen types I, V and their cross-links lysylpyridinoline, hydroxylysylpyridinoline and hydroxyproline.

Radiotherapy can lead to a reduction of bone density with an increased risk of pathological fractures. Bisphosphonates may represent a preventive treatment option by increasing the density of anorganic bone mineral. Yet it is unknown how bisphosphonates act on irradiated collagen cross-links, which play an essential role for the mechanical stability of bone. The aim of this study was to evaluate the effects of zoledronate on bone collagens and their cross-links after irradiation. The right femur of 37 rats was irradiated with a single dose of 9.5 Gy at a high dose rate using an afterloading machine. Half of the rats (n=18) received additionally a single dose zoledronate (0.1 mg/kg body weight). Fourteen and 100 days after irradiation the femora were collected for histologic evaluation and determination of the collagen cross-links lysylpyridinoline, hydroxylysylpyridinoline, and hydroxyproline. The collagen types were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Fourteen days after treatment the lysylpyridinoline levels of all treatment groups were significantly lower compared to the untreated control. After 100 days, in the combined radiotherapy+zoledronate group significantly lower lysylpyridinoline values were determined (p=0.009). Radiotherapy and/or zoledronate did not change significantly the level of hydroxylysylpyridinoline. The concentration of hydroxyproline was 14 days after irradiation significantly higher in the combined treatment group compared to the control. No significant differences were observed 100 days after treatment. Zoledronate does not have the ability to restore the physiological bone collagen cross-link levels after radiotherapy. However, this would be necessary for regaining the physiological mechanical stability of bone after irradiation and therefore to prevent effectively radiation-induced fractures.

The effects of tacrolimus on colonic anastomotic healing in rats.

AIM: The aim of this experimental study is to investigate the effects of tacrolimus on colonic anastomotic healing after subcutaneous administration. MATERIALS AND METHODS: Forty Albino-Wistar male rats were divided into two groups, with two equal subgroups each. They all underwent colonic resection followed by a single-layer, inverted colon anastomosis and were injected subcutaneously with either 1 ml of 0.9% NaCl solution or tacrolimus (0.1 mg/kg body weight) depending on their group. Half of the rats were sacrificed on the fourth postoperative day, while the remaining half were sacrificed on the eighth postoperative day. Macroscopical and histological assessment was performed, while anastomotic bursting pressures and the tissue concentrations in hydroxyproline and collagenase I were evaluated. RESULTS: On the fourth postoperative day, the bursting pressures (217.00 ± 11.12, p < 0.001), the fibroblast activity (2.80 ± 0.42, p = 0.022), the neoangiogenesis (2.10 ± 0.32, p = 0.007) and the tissue hydroxyproline concentration (254.23 ± 67.10, p = 0.001) were significantly higher in the tacrolimus-treated animals. Furthermore, tacrolimus significantly decreased the inflammatory cell infiltration (1.50 ± 0.53, p < 0.001) and the tissue collagenase I concentration (4.16 ± 0.76, p = 0.002). On the eighth day, the bursting pressure (264.00 ± 32.61, p < 0.001) and the hydroxyproline tissue concentration (331.04 ± 55.56, p = 0.002) were significantly higher in the tacrolimus subgroups. The inflammatory cell infiltration (1.20 ± 0.42, p < 0.001) and the collagenase I concentration (1.61 ± 0.83, p < 0.001) were significantly lower. In addition, the adhesion formation score was significantly lower (1.20 ± 0.92, p = 0.065). CONCLUSION: Tacrolimus, when injected subcutaneously, promotes healing of colonic anastomoses in rats. It impairs not only inflammatory response but also collagen degradation, resulting to increased anastomotic strength on the fourth as well as on the eighth postoperative day.

Intraperitoneally administered irinotecan with 5-fluorouracil impair wound healing of colonic anastomoses in a rat model: an experimental study.

AIM: The aim of this experimental study is the assessment of the effects of the immediate post-operative intraperitoneal administration of 5-fluorouracil and irinotecan on the healing process of large bowel anastomoses in rats. MATERIALS AND METHODS: Sixty male Wistar rats were divided into 4 groups of 15 rats each. The rats underwent large bowel resection and anastomosis, followed by the intraperitoneal administration of normal saline (group 1), 5-fluorouracil (group 2), irinotecan (group 3) or the combination of 5-fluorouracil and irinotecan (group 4). All animals were killed on the eighth post-operative day. During post-mortem examination, the anastomoses were assessed macroscopically for a possible anastomotic leak and the extent of adhesion formation. Subsequently, the anastomotic bursting pressure was measured, and the anastomoses were assessed histologically. RESULTS: No anastomotic dehiscence was observed in the rats of group 1. In groups 2 and 3, we observed 3 anastomotic leaks in each group, and in group 4, we observed 5 leaks (P = 0.111). The mean bursting pressure of the anastomoses in group 1 was significantly higher compared to groups 2, 3 and 4 (P < 0.001). The least inflammatory cell infiltration score was observed in group 1 (P < 0.001). The lowest neoangiogenesis score was observed in group 2 and the highest in group 4. The collagen formation in group 1 was significantly higher compared to the other 3 groups (P < 0.001). Similar results were observed for the fibroblast activity, where group 1 revealed significantly higher fibroblast scores compared to groups 2, 3 and 4 (P < 0.001). Finally, groups 2, 3 and 4 showed significantly lower hydroxyproline levels compared to the control group (P < 0.001). CONCLUSION: The immediate, post-operative intraperitoneal administration of 5-fluorouracil or irinotecan had a negative effect on the healing process of the large bowel anastomoses in rats. The negative effects of the combination of 5-fluorouracil and irinotecan were statistically more significant compared to the single use of 5-fluorouracil or irinotecan.

The effects of the intraperitoneal administration of oxaliplatin and 5-FU on the healing of colonic anastomoses: an experimental study.

BACKGROUND: The purpose of this experimental study was to assess the effects of the immediate postoperative intraperitoneal administration of oxaliplatin and 5-FU on the healing of colonic anastomoses in rats. METHODS: Sixty rats were randomized into 4 groups of 15 rats each and were subjected to colonic anastomoses. To the 1st group, saline solution was administered immediately postoperatively, intraperitoneally. To the 2nd group, 5-FU was administered, to the 3rd group oxaliplatin and to the 4th group 5-FU and oxaliplatin were administered immediately postoperatively, intraperitoneally. After killing the rats on the 8th postoperative day, the anastomoses were examined macroscopically and the anastomotic bursting pressures were measured. The anastomoses were also examined histologically and the hydroxyproline contents were determined. RESULTS: Rupture of the anastomosis was observed in no rats of the 1st group, in 3 rats of the 2nd group, in 4 rats of the 3rd group and in 7 rats of the 4th group (P = 0.016). The bursting pressure (P < 0.001), the hydroxyproline content (P < 0.001) and the concentration of collagen (P < 0.001) and fibroblasts (P < 0.001) were significantly lower in the 2nd, 3rd and 4th group in comparison with the 1st group. The formation of adhesions and the leukocytosis on the anastomoses were significantly higher in the 2nd, 3rd and 4th group than in the 1st group (P < 0.001). CONCLUSIONS: The immediate postoperative, intraperitoneal administration of oxaliplatin, 5-FU or the combination of 5-FU and oxaliplatin impairs the healing of colonic anastomoses in rats.

The effects of iloprost on colonic anastomotic healing in rats.

PURPOSE: The purpose of this experimental study is to investigate the effects of iloprost on colonic anastomotic healing in rats, after intraperitoneal administration. METHODS: Forty male Albino-Wistar rats were randomized into two groups of twenty animals each. They all underwent colonic resection followed by an inverted anastomosis. The rats of Group A (control) received 3 ml of NaCl intraperitoneally, while those of Group B (iloprost) received iloprost (2 µg/kg body weight), immediately postoperatively and daily until killed. Each group was further divided into two equal subgroups, depending on the day of killing. The animals of subgroups 1 were killed on the fourth postoperative day, while those of subgroups 2 on the eighth. Macroscopical and histological assessments were performed. Besides, anastomotic bursting pressures and the tissue concentrations in hydroxyproline and collagenase I were also evaluated. RESULTS: No anastomotic dehiscence was noted. The mean bursting pressure was higher in the iloprost group compared with the control group, but a significant difference was revealed only on the fourth postoperative day. Furthermore, iloprost significantly increased the new vessel formation on the fourth, as well as on the eighth postoperative day. CONCLUSION: Iloprost enhances the early phase of colonic anastomotic healing in rats.

[Diabetes mellitus ulcers treatment with Bletilla striata polysaccharide].

The aim of this study was to evaluate the efficacy of Bletilla striata polysaccharide on diabetes mellitus ulcers. Diabetes mellitus animal model was established by single ip injection of streptozotocin (STZ, 50 mg x kg(-1)) with the criteria of blood glucose > or = 16.7 mmol x L(-1) after 72 h. 4 weeks after STZ injection, each animal received two full thickness incisional wounds (1.8 cm in diameter). The wounds then were divided into B. striata polysaccharide group and PBS group. Wound closure rate, fibroblast (FB) infiltration, hydroxyproline (OHP) content and myeloperoxidase (MPO) levels were examined on day 3, 7, 14, 21 post wound. The treatment of B. striata polysaccharide significantly facilitated diabetes mellitus ulcers healing compared to PBS group. Histological analysis showed that B. striata polysaccharide markedly increased inflammatory cell infiltration in wound area. The herb also strongly evaluation of FB, OHP demonstrated a significantly increased in B. striata polysaccharide group. B. striata polysaccharide group promoted wound closure by means of enhanced inflammatory cell infiltration and re-epithelialization, and the promotion of FB and OHP levels.

Protective effect of quercetin against paraquat-induced lung injury in rats.

AIMS: Paraquat (PQ) is known to induce pulmonary injury via a redox cyclic reaction. The present study was aimed to determine the protective effects of quercetin against PQ-induced pulmonary injury in association with its antioxidant activity. MAIN METHODS: Male rats were challenged acutely by PQ (50 mg/kg, i.p.) with or without quercetin post-treatment. Pulmonary heme oxygenase-1 (HO-1) expression, malondialdehyde (MDA) level, and the total oxyradical scavenging capacity (TOSC) toward hydroxyl, peroxyl radicals and peroxynitrite were measured 24 h after PQ treatment. Different groups of rats were instilled with PQ (0.5 mg/kg) directly into the right lung. Quercetin was administered to the rats daily for 14 days after PQ instillation. Serum NO, pulmonary glutathione (GSH) and 4-hydroxyproline (4-HP) concentrations were quantified in conjunction with histopathological examination to determine the fibrotic changes in lung. KEY FINDINGS: Pulmonary MDA level and HO-1 expression were elevated and the TOSC was reduced rapidly by an intraperitoneal dose of PQ. These changes were inhibited by quercetin post-treatment. In rat lungs instilled with PQ 14 days before, NO, MDA and 4-HP were elevated, and GSH was reduced, which were all inhibited significantly by daily quercetin treatment. Histopathological examination also revealed that quercetin ameliorated the increase in fibroblast distribution and collagen deposition in the lungs instilled with PQ. SIGNIFICANCE: The present results demonstrate that quercetin administration to rats effectively inhibits the development of PQ-induced pulmonary injury most probably via its antioxidant activity.

Taurine attenuates radiation-induced lung fibrosis in C57/Bl6 fibrosis prone mice.

INTRODUCTION: The amino acid taurine has an established role in attenuating lung fibrosis secondary to bleomycin-induced injury. This study evaluates taurine's effect on TGF-beta1 expression and the development of lung fibrosis after single-dose thoracic radiotherapy. METHODS: Four groups of C57/Bl6 mice received 14 Gy thoracic radiation. Mice were treated with taurine or saline supplementation by gavage. After 10 days and 14 weeks of treatment, TGF-beta1 levels were measured in serum and bronchoalveolar lavage fluid (BALF). Lung collagen content was determined using hydroxyproline analysis. RESULTS: Ten days post radiotherapy, serum TGF-beta1 levels were significantly lower after gavage with taurine rather than saline (P = 0.033). BALF TGF-beta1 at 10 days was also significantly lower in mice treated with taurine (P = 0.031). Hydroxyproline content was also significantly lower at 14 weeks in mice treated with taurine (P = 0.020). CONCLUSION: This study presents novel findings of taurine's role in protecting from TGF-beta1-associated development of lung fibrosis after thoracic radiation.

The effect of cyanoacrylate in esophagocutaneous leakages occurring after esophageal anastomosis.

OBJECTIVE: Esophageal leakage (EL) continues to be a challenging pediatric surgical problem. The aim of this study was to investigate the effect of cyanoacrylate (Cy) in EL followed esophageal repair (ER). METHODS: Twelve rabbits were divided into control (C) and leakage (L) groups. A 1cm-length transverse esophageal incision at the level of the cervical region was made. In both groups, feeding was started orally 24h after the surgery for leakage creation. On postoperative day 7, primary repair was carried out in the C group and Cy instillation was performed in the L group. Esophagographic analysis was carried out on postoperative day 9 and the animals were fed orally on the same day on the condition that there was no esophageal leakage. The rabbits were sacrificed to measure diameters of the OR line, bursting pressure (BP), and hydroxyproline (HP) levels in the repaired cervical esophageal segment (RCES) 2 months later. RESULTS: The values of BP and HP in the C group were significantly higher than those in the L group. The diameters of the OR line in the L group were significantly greater compared to those in the C group. CONCLUSIONS: Cy glue instillation seems to be the ideal treatment for esophageal anastomosis leakages as shown by increased diameters of the OR line and decreased HP levels.

Preventive effects of curcumin on different aspiration material-induced lung injury in rats.

PURPOSE: We have studied whether curcumin protects different pulmonary aspiration material-induced lung injury in rats. MATERIALS AND METHODS: The experiments were designed in 60 Sprague-Dawley rats, randomly allotted into one of six groups (n=10): normal saline (NS, control), enteral formula (Biosorb Energy Plus, BIO), hydrochloric acid (HCl), NS+curcumin-treated, BIO+curcumin-treated, and HCl+curcumin-treated. NS, BIO, HCl were injected in to the lungs. The rats received curcumin twice daily only for 7 days. Seven days later, both lungs in all groups were examined histopathologically, immunohistochemically, and biochemically. Histopathologic examination was performed according to the presence of peribronchial inflammatory cell infiltration, alveolar septal infiltration, alveolar edema, alveolar exudate, alveolar histiocytes, interstitial fibrosis, granuloma, and necrosis formation. Immunohistochemical assessments were examined for the activity of inducible nitric oxide synthase (iNOS) and the expression of surfactant protein D (SP-D). Malondialdehyde (MDA), hydroxyproline (HP), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) activity were measured in the lung tissue. RESULTS: Our findings show that curcumin inhibits the inflammatory response reducing significantly (P<0.05) all histopathological parameters in different pulmonary aspiration models. Pulmonary aspiration significantly increased the tissue HP content, MDA levels and decreased the antioxidant enzyme (SOD, GSH-Px) activities. Curcumin treatment significantly decreased the elevated tissue HP content, and MDA levels and prevented inhibition of SOD, and GSH-Px enzymes in the tissues. Furthermore, our data suggest that there is a significant reduction in the activity of iNOS and a rise in the expression of SP-D in lung tissue of different pulmonary aspiration models with curcumin therapy. CONCLUSION: Our findings support the use of curcumin as a potential therapeutic agent in acute lung injury.

A TSP-1 synthetic peptide inhibits bleomycin-induced lung fibrosis in mice.

Bleomycin showed toxicity to lung and was recognized to induce a well model of lung fibrosis. Activated alveolar macrophages released increased amounts of transforming growth factor-beta1(TGF-beta1) in response to bleomycin-induced lung injury. Thrombospondin-1(TSP-1) was involved in the activation of latent TGF-beta1(L-TGF-beta1) through the association of the TSP-1/L-TGF-beta1 complex with the cell receptor of TSP-1, CD36. The antagonistic effects of the synthetic peptides were studied by the administration of TSP-1 (447-452) synthetic peptides to the mouse model. The hydroxyproline contents of the TSP-1-treated groups were significantly lower than those of other experimental groups. Inflammation, fibrotic degree and distribution of collagen fibers in the interstitial and alveolar in the TSP-1-treated groups were less than those of the other experimental groups. The expressions of collagen I and III in TSP-1-treated groups were significantly lower than in the other experimental groups. TSP-1 synthetic peptide reduced the tissue fibrotic pathologies and collagen accumulation in the model, resulting in the decreased severity of bleomycin-induced lung injury.

Metallothionein attenuates carmustine-induced oxidative stress and protects against pulmonary fibrosis in rats.

The present study was carried out to evaluate the effect of exogenously administered metallothionein (MT) against carmustine (BCNU)-induced lung toxicity in rats. A total of 60 rats were randomly divided into four groups (15/group): control group in which the animals received 0.5 ml physiologic saline containing 10% ethanol (IP) weekly, MT-administered group in which rats received MT (30 micromol/kg, IP) weekly, BCNU-administered group in which rats received BCNU (5 mg/kg, IP) weekly and MT + BCNU group in which rats received weekly doses of BCNU (5 mg/kg, IP) followed 24 h later by MT (30 micromol/kg, IP). At the end of the experiment (after 6 weeks), lung histological changes, collagen staining, the activity of glutathione reductase (GR) and contents of reduced glutathione (GSH) and hydroxyproline (Hpr) in the lung as well as serum level of tumor necrosis factor-alpha (TNF-alpha) were evaluated. The obtained data revealed that BCNU induced pathological changes and markedly increased lung collagen and level of Hpr but decreased GSH content and GR activity and increased serum TNF-alpha compared to both control and MT-administered rats. Administration of MT + BCNU markedly improved histological features and decreased staining of collagen along with increased GR activity, GSH content but decreased level of Hpr in lung tissue as well as decreased serum level of TNF-alpha compared with BCNU-treated rats. Based on our results, it is possible to postulate that exogenous MT can act against BCNU-induced lung toxicity by a mechanism related, at least in part, to its ability to decrease oxidative stress and fibrosis.

Aquilegia vulgaris extract attenuates carbon tetrachloride-induced liver fibrosis in rats.

Six groups of male Wistar rats were treated as follows: in groups II, III and V liver damage was induced by CCl(4) (per os, 1590 mg/kg b.w.day) given 2 days a week for 6 weeks; group III was treated simultaneously with ethanol extract of Aquilegia vulgaris (100 mg/kg b.w.day) for 6 weeks; group V with silymarin, positive control, at a dose of 100 mg/kg b.w.day for 6 weeks; and groups IV and VI received only the extract or silymarin, respectively. Microsomal lipid peroxidation in the liver increased following CCl(4) treatment by 61-213% and was not changed significantly by the extract. The effect of silymarin was more pronounced, 19-52% decrease in the lipid peroxidation level. Hepatic glutathione was depleted by 22% in CCl(4)-treated rats. The extract tested did not change this parameter. The activity of antioxidant enzymes was significantly reduced after CCl(4) administration, by 42-63%. Co-administration of the extract or silymarin resulted in significant increase in these enzymes activity; however, the basal level was not reached. Hepatic hydroxyproline concentration was elevated over 5-fold in comparison with controls. Co-administration of the extract or silymarin decreased the level of hydroxyproline by 66% and 55%, respectively. Activity of serum hepatic enzymes was elevated in rats treated with CCl(4) by 47-8700%. Both the extract and silymarin reduced significantly these enzymes' activity. The extract caused a fall in bilirubin and cholesterol level in rats treated with CCl(4) by 42% and 17%, respectively. Histopathological examination revealed less-severe fibrosis in rats co-administered the extract or silymarin when compared to animals treated with CCl(4) alone.