Protective Effects Of Rosmarinic Acid Research Articles

Rosmarinic acid alleviated intestinal barrier damage caused by Escherichia coli by regulating the gut microbiota and inhibiting the NF-κB signalling pathway in mice.

Escherichia coli (E. coli) is a common zoonotic foodborne pathogen that poses a major threat to public health and economic development. Rosmarinic acid (RA) can inhibit intestinal inflammation; however, the protective effect of RA against the intestinal barrier damage induced by E. coli in mice and the underlying mechanism have not been elucidated. In this study, mice were orally administered with RA (20 mg kg-1) by gavage for one week and then were intraperitoneally challenged with E. coli. Mouse colonic epithelial cells (MCECs) were pretreated with RA for 6 h and challenged with E. coli (MOI = 1000) for 3 h. The results revealed that RA alleviated E. coli-induced weight loss in mice; reduced the increase in the levels of TNF-α, IL-6 and IL-1β in the serum; alleviated the decrease in ZO-1 protein expression; and increased intestinal permeability by inhibiting the NF-κB signalling pathway both in vivo and in vitro. Moreover, RA relieved the increase in intestinal permeability, reversed the structural damage to the mouse gut microbiota caused by E. coli, and increased the abundance of beneficial bacteria, including Lachnospiraceae_NK4136_group. Additionally, RA lost its protective function against E. coli infection in a pseudosterile mouse model, suggesting that the protection induced by RA was dependent on the gut microbiota. In conclusion, these results indicate that RA alleviates E. coli-induced inflammatory damage to the intestinal barrier by inhibiting the NF-κB signalling pathway and maintaining gut microbiota homeostasis. These findings provide new ideas and foundations for the application of RA as protection against E. coli.

Protective effects of rosmarinic acid against autistic-like behaviors in a mouse model of maternal separation stress: behavioral and molecular amendments.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder with worldwide increasing incidence. Maternal separation (MS) stress at the beginning of life with its own neuroendocrine changes can provide the basis for development of ASD. Rosmarinic acid (RA) is a phenolic compound with a protective effect in neurodegenerative diseases. The aim of this study was to determine the effect of RA on autistic-like behaviors in maternally separated mice focusing on its possible effects on neuroimmune response and nitrite levels in the hippocampus. In this study, 40 mice were randomly divided into five groups of control (received normal saline (1 ml/kg)) and MS that were treated with normal saline (1 ml/kg) or doses of 1, 2, and 4 mg/kg RA, respectively, for 14 days. Three-chamber sociability, shuttle box, and marble burying tests were used to investigate autistic-like behaviors. Nitrite level and gene expression of inflammatory cytokines including TNF-α, IL-1β, TLR4, and iNOS were assessed in the hippocampus. The results showed that RA significantly increased the social preference and social novelty indexes, as well as attenuated impaired passive avoidance memory and the occurrence of repetitive and obsessive behaviors in the MS mice. RA reduced the nitrite level and gene expression of inflammatory cytokines in the hippocampus. RA, probably via attenuation of the nitrite level as well as of the neuroimmune response in the hippocampus, mitigated autistic-like behaviors in maternally separated mice.

The ameliorative effect of rosmarinic acid and epigallocatechin gallate against doxorubicin-induced genotoxicity

Doxorubicin (Dox), an effective anticancer agent, is known for its genotoxic effects on normal cells. Phenolic compounds, renowned for their antitumor, antioxidant, and antigenotoxic properties, have gained prominence in recent years. This study investigates the individual and combined protective effects of rosmarinic acid (RA) and epigallocatechin gallate (EGCG) against Dox-induced genotoxicity using various in vitro test systems. The synergistic/antagonistic interaction of these combinations on Dox’s chemotherapeutic effect is explored in breast cancer cell lines. Both RA and EGCG significantly mitigate Dox-induced genotoxicity in comet, micronucleus, and Ames assays. While Dox exhibits higher selectivity against MCF-7 cells, EGCG and RA show greater selectivity against MDA-MB-231 cells. The coefficient of drug interaction reveals a synergistic effect when RA or EGCG is combined with Dox in breast cancer cells. In conclusion, both EGCG and RA effectively reduce Dox-induced genetic damage and enhance Dox’s cell viability-reducing effect in breast cancer cells.

Rosmarinic Acid Liposomes Downregulate Hepcidin Expression via BMP6-SMAD1/5/8 Pathway in Mice with Iron Overload.

The objective of this study is to examine the potential protective effect of rosmarinic acid (RosA) encapsulated within nanoliposomes (RosA-LIP) on hepatic damage induced by iron overload. The characteristics, stability, and release of RosA-LIP in vitro were identified. The mice were randomly assigned to five groups: Control, Model, Model+DFO (DFO), Model+RosA (RosA), and Model+RosA-LIP (RosA-LIP). The iron overload model was induced by administering iron dextran (i.p.). The DFO, RosA, and RosA-LIP groups received iron dextran and were subsequently treated with DFO, RosA, and RosA-LIP for 14 days. We developed a novel formulation of RosA-LIP that exhibited stability and controlled release properties. Firstly, RosA-LIP improved liver function and ameliorated pathological changes in a mouse model of iron overload. Secondly, RosA-LIP demonstrated the ability to enhance the activities of T-SOD, GSH-Px, and CAT, while reducing the levels of MDA and 4-HNE, thereby effectively mitigating oxidative stress damage induced by iron overload. Thirdly, RosA-LIP reduced hepatic iron levels by downregulating FTL, FTH, and TfR1 levels. Additionally, RosA-LIP exerted a suppressive effect on hepcidin expression through the BMP6-SMAD1/5/8 signaling pathway. Furthermore, RosA-LIP upregulated FPN1 expression in both the liver and duodenum, thereby alleviating iron accumulation in these organs in mice with iron overload. Notably, RosA exhibited a comparable iron chelation effect, and RosA-LIP demonstrated superior efficacy in mitigating liver damage induced by excessive iron overload. RosA-LIP exhibited favorable sustained release properties, targeted delivery, and efficient protection against iron overload-induced liver damage. A schematic representation of the proposed protective mechanism of rosmarinic acid liposome during iron overload. Once RosA-LIP is transported into cells, RosA is released. On the one hand, RosA attenuates the BMP6-SMAD1/5/8-SMAD4 signaling pathway activation, leading to inhibiting hepcidin transcription. Then, the declined hepcidin contacted the inhibitory effect of FPN1 in hepatocytes and duodenum, increasing iron mobilization. On the other hand, RosA inhibits TfR1 and ferritin expression, which decreases excessive iron and oxidative damage.

Rosmarinic Acid Alleviates Radiation-Induced Pulmonary Fibrosis by Downregulating the tRNA N7-Methylguanosine Modification-Regulated Fibroblast-to-Myofibroblast Transition Through the Exosome Pathway.

Radiation-induced pulmonary fibrosis (RIPF) is a common complication after radiotherapy in thoracic cancer patients, and effective treatment methods are lacking. The purpose of this study was to investigate the protective effect of rosmarinic acid (RA) on RIPF in mice as well as the mechanism involved. m7G-tRNA-seq and tRNA-seq analyses were conducted to identify m7G-modified tRNAs. Western blotting, immunohistochemistry, northwestern blotting, northern blotting, immunofluorescence, wound-healing assays and EdU experiments were performed to explore the molecular mechanism by which RA regulates fibroblast-to-myofibroblast transformation (FMT) by affecting the exosomes of lung epithelial cells. Ribo-seq and mRNA-seq analyses were used to explore the underlying target mRNAs. Seahorse assays and immunoprecipitation were carried out to elucidate the effects of RA on glycolysis and FMT processes via the regulation of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) acetylation. We found that RA had an antifibrotic effect on the lung tissues of RIPF model mice and inhibited the progression of FMT through exosomes derived from lung epithelial cells. Mechanistically, RA reduced the transcription and translation efficiency of sphingosine kinase 1 in lung fibroblasts by decreasing N7-methylguanosine modification of tRNA, downregulating the expression of tRNAs in irradiated lung epithelial cell-derived exosomes, and inhibiting the interaction between sphingosine kinase 1 and the N-acetyltransferase 10 protein in fibroblasts. Furthermore, the acetylation and cytoplasmic translocation of PFKFB3 were reduced by exosomes derived from irradiated lung epithelial cells, which following RA intervention. This suppression of the FMT process, which is triggered by glycolysis, and ultimately decelerating the progression of RIPF. These findings suggest that RA is a potential therapeutic agent for RIPF.

Rosmarinic acid alleviates intestinal inflammatory damage and inhibits endoplasmic reticulum stress and smooth muscle contraction abnormalities in intestinal tissues by regulating gut microbiota.

The host-bacterial interactions play the key role in inflammatory bowel disease (IBD). Dysbiosis of the intestinal flora can lead to pathological changes in the intestine. Rosmarinic acid (RA) is a natural phenolic acid compound with antioxidant, anti-cancer, anti-inflammatory, anti-apoptotic, anti-fibrotic, and anti-bacterial activities that has a palliative effect on acute IBD. We have established an in vivo model for mice. Histological staining was performed to directly observe RA alterations in the intestinal tract. The alteration of RA on mouse intestinal flora was observed by 16S rRNA high-throughput sequencing, and the effect of RA on intestinal mechanism of action was detected by qPCR and western blot. The results showed that RA had a significant protective effect on the intestine. RA upregulated the abundance of Lactobacillus johnsonii and Candidatus Arthromitus sp SFB-mouse-NL and downregulated the abundance of Bifidobacterium pseudolongum, Escherichia coli, and Romboutsia ilealis. RA downregulated the expressions of ROCK, RhoA, CaM, MLC, MLCK, ZEB1, ZO-1, ZO-2, occludin, E-cadherin, IL-1β, IL-6, TNF-α, GRP78, PERK, IRE1, ATF6, CHOP, Caspase12, Caspase9, Caspase3, Bax, Cytc, RIPK1, RIPK3, MLKL, and upregulated the expression of IL-10 and Bcl-2. These results displayed that RA inhibited the inflammation, which is caused by tight junction damage, by repairing intestinal flora dysbiosis, relieved endoplasmic reticulum stress, inhibited cell death, and corrected smooth muscle contractile dysregulation. The results of this study revealed RA could have a protective effect on the small intestine of mice by regulating intestinal flora. IMPORTANCE Inflammatory bowel disease (IBD) is a chronic, relapsing, remitting disorder of the gastrointestinal system. In this study, we investigated the protective effects of rosmarinic acid on the intestinal tract. The results showed that RA was effective in reducing inflammatory damage, endoplasmic reticulum stress, smooth muscle contraction abnormalities, and regulating intestinal flora disorders.

Investigation of the protective effect of rosmarinic acid in rats given high dose gentamicin

Abstract
 Purpose: Gentamicin is an antibiotic, used to treat infections caused by microorganisms. Endogenous antioxidants are strengthened by using antioxidant protectors against the toxic effects caused by drugs. Rosmarinic acid has antioxidative, anti-inflammatory, antiapoptotic, and antitumoral effects. In our study, we aimed to investigate the protective effect, immunohistochemical changes, oxidative markers, histopathological changes and inflammation related Ifi44 expression of rosmarinic acid against gentamicin induced nephrotoxicity in rats.
 Materials and methods: In our study, 32 rats randomly divided into four groups. These groups are consisted of control group, gentamicin group received gentamicin 100 mg/kg/day, gentamicin + rosmarinic group acid received gentamicin 100 mg/kg/day and rosmarinic acid 50 mg/kg/day, and rosmarinic acid group received rosmarinic acid 50 mg/kg/day. At the end of the study, histopathological, immunohistochemical, and biochemical changes in kidney tissues evaluated. 
 Results: Rosmarinic acid reduced creatine, urea, blood urea nitrogen, and total oxidative stress in blood serum. The toxic effect of gentamicin caused severe histopathological changes in the kidneys. A slight decrease in histopathological changes observed in the gentamicin + rosmarinic acid group. Antiproliferative Ifi44 expression was higher in the gentamicin group and gentamicin + rosmarinic acid group.
 Conclusion: As a result of the application of rosmarinic acid, a decrease in oxygen radicals and an increase in antioxidant levels observed. When used in combination with gentamicin and rosmarinic acid, the protective effect of rosmarinic acid was partially observed, but it could not provide full protection.

Protective effect of rosmarinic acid on the transmembrane transporter Ctr1 expression in cisplatin-treated mice.

Cisplatin (cis-diamminedichloroplatinum(II), CP) is a platinum-based anticancer drug widely used in the treatment of solid malignancies. However, its side effects, particularly nephrotoxicity, are limiting factors in its clinic use. Rosmarinic acid (RA), a natural antioxidant compound, is reported to attenuate oxidative stress and associated pathophysiological outcomes. Our study aimed to explore the protective effect of RA against CP-induced acute kidney injury (AKI). We investigated the effect of RA at the dose of 100 mg/kg on AKI induced by CP (20 mg/kg) in mice. Various parameters of nephrotoxicity such as levels of serum electrolytes, albumin, and globulin were measured using standardized methods. Besides, a specific biomarker of damage to proximal tubular cells, kidney injury molecule-1 (Kim-1), was measured in the serum by ELISA. mRNA expression of Kim-1 and a transmembrane transporter, copper transporter 1 (Ctr1), was analyzed by quantitative reverse transcriptase-polymerase chain reaction. CTR1 expression was also analyzed by western blot technique. RA treatment restored the downregulated CTR1 , a renal transmembrane transporter in CP-treated mice. It was accompanied by a reduction in the level of serum albumin and globulin. Serum electrolytes such as Na+, K+, and Ca2+ in CP-treated mice were found to be restored with RA treatment. Moreover, RA also significantly downregulated the increased expression of nephrotoxicity biomarker KIM-1. Overall, RA proved to be an effective nephroprotective compound which afforded protection at cellular and subcellular levels with an appreciable modulatory effect on a transmembrane transporter.

Protective Efficacy of Rosmarinic Acid on Acute Pancreatitis in Rats

Abstract
 Aim: Acute pancreatitis is a serious disease, with an incidence of 5 - 35 in 100,000 individual. New studies are constantly planned for the treatment of pancreatitis. Many studies have shown that Rosmarinic acid has antioxidant properties. In this study, we examined the protective effect of Rosmarinic acid on acute pancreatitis.
 Material and Methods: A total of 28 animals were used during the experiment, and 4 groups were formed with 7 animals in each group. Group 1 is the control group. The rats in Group 2 were administered 75 μg/kg Cerulein every hour intraperitoneally at one hour intervals, a total of four times. Group 3 experimental animals were given 50 mg/kg Rosmarinic acid by per oral gavage. The rats in group 4 were given 50 mg/kg Rosmarinic acid per oral gavage after 75 μg/kg Cerulein was injected intraperitoneally every hour for a total of four times. Afterwards, all animals were sacrificed by exsanguination, blood samples and pancreatic tissue were taken for examination.
 Results: Examination of pancreatic tissues revealed necrosis, edema and inflammation in the acute pancreatitis group. Both histopathological and serum values of the rosmarinic acid group were close to the control group. The use of Rosmarinic acid after acute pancreatitis had a positive effect on the pacreatic tissues and blood values, but still did not cause complete recovery.
 Conclusion: In the case of acute pancreatitis, it was concluded that rosmarinic acid has a partial curative effect, but still does not provide a full recovery.

Protective Effect of Rosmarinic Acid on Influenza Virus-induced Pneumonia in Mice

Protective Effect of Rosmarinic Acid on Influenza Virus-induced Pneumonia in Mice

Investigation of the Effect of Rosmarinic Acid on Cyclophosphamide-Induced Gonadal Toxicity

The aim of this study was to investigate the effect of rosmarinic acid against gonadal toxicity caused by cyclophosphamide, an important anticancer drug. A total of 28 rats were divided into 4 groups, with 7 animals in each group. The groups were created as follows; Group 1 (control) (n = 7): Subjects received only 1 ml of 0.9% saline solution per day intraperitoneally for 14 days. Group 2 (Rosmarinic Acid) (n = 7): The subjects were given 20 mg/kg Rosmarinic acid intraperitoneally for 14 days. Group 3 (Cyclophosphamide) (n = 7): Only 1 ml of 0.9% saline solution was administered intraperitoneally to the subjects for the first 7 days. Cyclophosphamide 20 mg/kg per day was administered intraperitoneally for the last 7 days (from the 8th day). Group 4 (Rosmarinic Acid + Cyclophosphamide) (n = 7): The subjects were given 20 mg/kg Rosmarinic acid daily intraperitoneally for 14 days. From the 8th day of the experiment to the end of the experiment (last 7 days of the experiment), 20 mg/kg cyclophosphamide was given intraperitoneally daily. At the end of the experiment, body weights of all rats were measured first. Afterwards, the weights of testicular tissue samples were measured and the averages of the weights were taken. Routine tissue follows up was performed on the testicular tissues taken. Hematoxylin-eosin staining was applied to tissue sections of 5 μm thickness. As a result of the statistical analysis, it was determined that cyclophosmadine decreased body and testicular weight, but rosmarinic acid had a protective effect on the contrary. It was concluded that cyclophosmadine caused damage to the basement membrane structure, Sertoli, Leydig and germ cells, but these structures were preserved due to the protective effect of rosmarinic acid. Despite the toxic effect of CP, rosmarinic acid is thought to have a significant curative effect on the spermatogenetic process and seminiferous tubule structure in the gonads.

Protective effects of rosmarinic acid against azoxymethane-induced colorectal cancer in rats.

Colorectal cancer (CRC) incidence is increasing gradually and has been become one of the most common cancers worldwide. Hence, it is important to discover cheap, naturally occurring compounds to be effective in suppressing the devastating effect of colon-related tumors. Rosmarinic acid (RA), one of the compounds of plant origin, possesses attractive features for use as an agent for cancer prevention and treatment. This study investigated the ability of RA to prevent azoxymethane (AOM)-induced rat colon carcinogenesis by evaluating the effect of RA on tumor formation and circulatory oxidant-antioxidant status. Moreover, plasma levels of adiponectin (APN) monocyte chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6) were detected by enzyme linked immunosorbent assay. The animals were divided into three groups: Control, AOM, and AOM + RA. Rats were fed a modified pellet diet (15.8% peanut oil was added to the standard diet) during the experimental period. Colon cancer was formed by applying 15 mg/kg AOM intraperitoneal once a week for 4 weeks in both the CRC group and AOM + RA group. Besides AOM, AOM + RA group received 5 mg/kg body weight RA orally every day during the study. The results showed that adenocarcinoma rates formed87.5% ofthe AOM group. With treatment of RA, a reduction in the incidence of adenocarcinoma was observed in the AOM + RA group. The plasma MCP-1, IL-6, and TO levels were significantly higher, APN and TAS levels were significantly lower in the AOM group with respect tocontrols. In addition, there was a significant increase in TAS levels in the RA treatment group compared to the AOM group. These findings suggested that RA may be beneficial in preventing AOM-induced colon carcinogenesis formation.

In Vivo and In Vitro Protective Effects of Rosmarinic Acid against Doxorubicin-Induced Cardiotoxicity

Doxorubicin (DOX) is an anticancer medicine that may trigger cardiomyopathy. Rosmarinic acid (RA) has shown antioxidant, anti-inflammatory, and anticancer effects. This investigation assessed the cardioprotective effect of RA on DOX-induced-toxicity in both in vivo and in vitro experiments. Male rats were randomized on 7 groups: (1) control, (2) DOX (2 mg/kg, per 48 h, 12d, i.p), (3) RA (40 mg/kg, 12d, i.p.), (4–6) RA (10, 20, 40 mg/kg, 16d, i.p.)+ DOX, (7) Vitamin E (200 mg/kg, per 48 h, 16d, i.p.) + DOX and then indices of cardiac function were estimated. Also, DOX and rosmarinic acid effects were examined on MCF7 cells (breast cancer cells line) to clarify that both cardiotoxicity and anticancer effects were analyzed. DOX increased heart to body weight ratio, RRI, QA, STI, QRS duration and voltage, attenuated HR, blood pressure, Max dP/dt, Min dP/dt, LVDP, enhanced MDA, declined GSH amount, and caused fibrosis and necrosis in cardiac tissue. Administration of RA ameliorated the toxic effects of DOX. In vitro studies showed that RA did not affect the cytotoxic effect of DOX. RA as an antioxidant, anti-inflammatory, and cardioprotective compound could be a promising compound to help minimize DOX-induced cardiotoxicity.

The protective effects of rosmarinic acid on ethanol-induced gastritis in male rats: antioxidant defense enhancement.

Background and purpose:Gastritis is one of the most current gastrointestinal disorders worldwide. Alcohol consumption is one of the major factors, which provides gastritis. Rosmarinic acid (RA) is found in many plants and has powerful antioxidant and anti-inflammatory effects. In this study, the protective effect of RA was evaluated on the histopathological indices, antioxidant ability, and prostaglandin E2 (PGE2) secretion in male rats.Experimental approach:Forty-two animals were divided into control, ethanol-induced gastritis, and RA groups, 6 each. The protective groups included RA administration before gastritis induction at 50 mg (R-G50), 100 mg (R-G100), 150 mg (R-G150), and 200 mg (R-G200) doses. Gastritis was induced by gavage of 1 mL pure ethanol in fasted animals. After 1 h of gastritis induction, the rats were sacrificed and stomach tissue was removed.Findings/Results:Histological evaluation revealed that RA significantly attenuated gastric ulcers, leucocyte infiltration, and hyperemia. It also increased mucosal layer thickness and restored gastric glands. Furthermore, RA decreased malondialdehyde level, increased superoxide dismutase, catalase, and glutathione in the stomach tissue, and raised gastric PGE2 level.Conclusion and implications:Our study demonstrated that rosmarinic acid has a notable effect on gastritis protection that could be due to increased antioxidant defense and PGE2 secretion, eventually maintenance of mucosal barrier integrity and gastric glands.

Rosmarinic acid inhibits high glucose-induced cardiomyocyte hypertrophy by activating Parkin-mediated mitophagy

To evaluate the effect of rosmarinic acid (RA) on mitophagy and hypertrophy of cardiomyocytes exposed to high glucose (HG). Rat cardiomyocytes (H9c2) exposed to HG (25 mmol/L) were treated with 50 μmol/L RA or with both RA treatment and Parkin siRNA transfection, with the cells cultured in normal glucose (5.5 mmol/L) and HG as the controls. The expressions of PINK1, Parkin and LC3II/LC3I in the cells were detected by Western blotting. The formation of mitochondrial autophagosomes was observed by transmission electron microscope. Flow cytometry was employed to detect the level of reactive oxygen species (ROS) and apoptotic rate of the cells. The activities of respiratory chain complex enzymes were measured by spectrophotometry. Fluorescence enzyme labeling and 3H-leucine labeling were used for determining the level of membrane potential and protein synthesis rate, respectively. The cell surface area was observed by light microscopy. RA treatment significantly increased the expression levels of PINK1, Parkin and LC3-II/I (P < 0.05), promoted the formation of mitochondrail autophagosome, inhibited the production of reactive oxygen species (P < 0.05), restored the activities of mitochondrial respiratory chain complex enzymes and mitochondrial membrane potential (P < 0.05), inhibited apoptosis (P < 0.05), and reduced the cell surface area and protein synthesis rate of H9c2 cells induced by HG exposure (P < 0.05). The protective effects of RA against HG-induced oxidative stress and cardiomyocyte hypertrophy was obviously blocked by inhibition of mitophagy mediated by transfection with Parkin siRNA (P < 0.05). RA can protect rat cardiomyocytes against oxidative stress injury and cardiomyocyte hypertrophy induced by HG by activating Parkin-mediated mitophagy.

Rosmarinic acid inhibits oxLDL-induced inflammasome activation under high-glucose conditions through downregulating the p38-FOXO1-TXNIP pathway

Elevated glucose levels in diabetes mellitus is associated with increased oxidized low density lipoprotein (oxLDL). High glucose (HG) and oxLDL are key inducers of oxidative stress and inflammatory processes responsible for diabetic vascular disorders. Rosmarinic acid is a polyphenol with antioxidant, anti-inflammatory and insulin-sensitizing effects. However, whether rosmarinic acid protects against diabetic atherosclerosis remains unknown. In this study, we aimed to investigate the protective effect of rosmarinic acid against diabetic atherosclerosis and the related signaling pathway. oxLDL-mediated oxidative stress upregulated thioredoxin-interacting protein (TXNIP) and subsequently induced binding of TXNIP to NLRP3 to mediate NLRP3 inflammasome assembly and activation under HG conditions in ECs. Reactive oxygen species (ROS) scavengers, p38 and FOXO1 inhibitors and TXNIP siRNA inhibited TXNIP protein upregulation and NLRP3 inflammasome assembly and activation. Rosmarinic acid abrogated TXNIP protein upregulation and the interaction between TXNIP and NLRP3 to attenuate NLRP3 inflammasome assembly and activation and eventually IL-1β secretion in ECs through downregulating ROS production, p38 phosphorylation and FOXO1 protein induction in ECs. These findings show that rosmarinic acid inhibits endothelial dysfunction which is shown in diabetic atherosclerosis through downregulating the p38-FOXO1-TXNIP pathway and inhibiting inflammasome activation.

The protective effect of rosmarinic acid on myotube formation during myoblast differentiation under heat stress.

High ambient temperature is one of the most important environmental factors that caused the reduction of livestock productivity and the increase of mortality. It has been shown that heat stress could affect the meat quality characteristics by physiological and metabolic perturbations in live livestock. Rosmarinic acid (RA) is a natural polyphenolic phytochemical compound that has many important biological activities, such as antioxidant, antimutagenic, and antitumor. The purpose of this study was to investigate the possible function and mechanism of RA on myoblast proliferation and differentiation under heat stress condition. The results showed that heat stress reduced the viability of myoblast and increased the percentage of apoptotic cells, and it also disrupted myotube formation by altering the expression of myogenic regulatory factors MyoD, myogenin, and MyHC. However, pretreatment of RA can protect C2C12 cells from heat stress-induced apoptosis, and it also increased the expression level of MyoD, myogenin, and MyHC under heat stress, which indicated that RA have protective effect on heat stress-caused failure of myotube formation during myoblast differentiation. Above all, our finding demonstrated that RA can promote the differentiation of C2C12 myoblast and maintain the formation of myotubes even under heat stress condition.

Protective roles and mechanisms of rosmarinic acid in cyclophosphamide-induced premature ovarian failure.

The aim of this study was to investigate the protective effect of rosmarinic acid (RA) in a premature ovarian failure (POF) mouse model and the potential mechanisms. The POF model was induced by a single intraperitoneal injection of 120 mg/kg cyclophosphamide (CP). Additionally, 40 mg/kg RA was administered for 7 days before CP injection. The concentration of sex hormones was determined by fluorescence immunohistochemistry. Histological analysis was performed after ovarian tissue sections were stained with hematoxylin and eosin. The expression of the NLRP3 inflammasome was examined by western blot analysis and polymerase chain reaction. The expression of apoptosis markers of cytochrome c and caspase-3 was also detected by western blot analysis and immunohistochemistry. The results showed that RA not only decreased the ovarian index in POF mice but also improved the abnormal secretion of reproductive hormones associated with POF. Treatment with RA suppressed the ovarian expression of the NLRP3 inflammasome and regulated the ovarian expression of apoptosis-related proteins. The results suggested that RA exhibited a protective effect against CP-induced POF potentially by suppressing apoptosis and the NLRP3 inflammasome.

Investigation of antioxidant effects of rosmarinic acid on liver, lung and kidney in rats: a biochemical and histopathological study.

The aim of the study was to investigate the protective effects of rosmarinic acid in rats exposed to hepatic ischaemia/reperfusion (I/R) injury. Thirty-two rats were randomly classified into four groups of 8 rats each: laparotomy without medication, rosmarinic acid (dose of 50 mg/kg via oral gavage) followed by laparotomy, laparotomy followed by hepatic I/R, and hepatic I/R with rosmarinic acid. Serum aspartate aminotransferase, alaninę aminotransferase, and malondialdehyde levels and total oxidant activity and total antioxidant capacity levels of the liver, lung, and kidney were assessed. The histopathologic assessment was also performed. Rosmarinic acid significantly reduced liver function test parameters and decreased oxidative stress and abnormal histopathologic findings in the liver. The oxidative stress in the lung significantly increased in the I/R group but significantly decreased in the I/R + rosmarinic acid group due to the addition of rosmarinic acid. Rosmarinic acid led to no reduction in oxidative stress in kidney following hepatic I/R injury. There were no statistically significant differences among the groups regarding histopathologic changes in kidney and lung sections. Rosmarinic acid has antioxidant properties and is an effective hepatoprotective agent. However, although rosmarinic acid provides useful effects in the lung by increasing antioxidant capacity and reducing oxidative stress after I/R injury, it does not ameliorate histopathologic changes. These findings suggest that rosmarinic acid is likely to provide favourable outcomes in the treatment of hepatic I/R injury.

The Protective Effect of Rosmarinic Acid Against Unfavorable Influence of Methylparaben and Propylparaben on Collagen in Human Skin Fibroblasts.

Parabens, which are widely used in food, medicines and cosmetics, have a harmful effect on human health. People are most exposed to parabens transdermally by using cosmetic products containing these preservatives. The purpose of this study was to estimate the influence of parabens (methylparaben—MP and propylparaben—PP) on the metabolism of collagen in the human skin fibroblasts and above all, to assess whether rosmarinic acid (RA—50, 100, or 150 μM) can protect these cells from the adverse effects of parabens (0.001% MP and 0.0003% PP, 0.003% MP and 0.001% PP, and 0.01% MP and 0.003% PP). The possible mechanisms of RA action were estimated as well. Parabens decreased the expression of collagen type I and III at mRNA and protein levels, while RA (depending on the concentration) provided partial or total protection against these changes. The effective protection against the adverse effects of parabens on cell viability and proliferation was also provided by RA. The beneficial impact of RA on collagen and the fibroblasts resulted from an independent action of this compound and its interaction with parabens. This study allows us to conclude that this polyphenolic compound may protect from unfavorable health outcomes caused by lifetime human exposure to parabens contained in cosmetic products.