Transdermal fennel essential oil nanoemulsions with promising hepatic dysfunction healing effect: in vitro and in vivo study.

Fennel (Foeniculum vulgare Mill.) is a member of family Apiaceae. Trans-anethole, the major component of Fennel essential oil (FEO), possesses antioxidant and hepatoprotective effects. Transdermal nanoemulsions (NEs) are advanced colloidal systems for systemic and controlled drug delivery through the stratum corneum barrier. FEO NEs were prepared using the oil Lauroglycol™ 90, as it provides a larger NE existence zone than Captex® 300, in the constructed phase diagrams. Six systems were prepared using Tween20/propylene glycol (S/CoS) in the ratios 2:1 and 3:1 with oil to S/CoS mass ratios 1:9, 2:8 and 3:7. Physicochemical characterization revealed optimum properties regarding thermodynamic stability, droplet size and pH with a Newtonian flow pattern. In vitro permeation study in rat skin revealed the highest cumulative amount permeated (µg/cm2), flux and permeability coefficient values for F4 made up of 2% FEO, 4.67% Lauroglycol™ 90, 60% S/CoS in the ratio 3:1. Results of the in vivo hepatic dysfunction study in rats indicate promising significant amelioration of liver function reflected in ALT, AST, ALP, bilirubin, albumin, malondialdehyde and ammonia plasma levels. The results signify the promising approach of FEO NEs in achieving remedy of liver toxicity. The most promising effect is inherent to F4 which imparts a more positive effect than FEO.

Role of Probiotic Mixture with and Without Green Tea Extract in Prevention of Hepatorenal Syndrome in Rat Model.

BACKGROUND AND OBJECTIVE: Hepatorenal syndrome (HRS) is a major public health problem in which both liver and kidney dysfunctions are encountered. The present research aimed to investigate the beneficial use of micro-encapsulated probiotic alone (Bifidobacterium bifidum, Lactobacillus delbrueckii and Streptococcus thermophilus mixture) or with green tea alcohol extract in HRS model in rats. MATERIALS AND METHODS: Flavonoids content and in vitro antioxidant activity of the extract were assessed. The animal experiment consisted of 4 groups; control healthy, control with HRS and two test groups with HRS and treated with either the encapsulated probiotic mixture alone or with green tea extract. After 3 weeks; urinary creatinine was determined in 24 h rat urine samples. Colonic microbiota was assessed in faeces. Plasma malondialdehyde, nitrite, C-reactive protein, creatinine, uric acid, urea and the activity of transaminases, catalase (CAT) and angiotensin-1 converting enzyme (ACE-1) were determined with calculation of creatinine clearance. RESULTS: Results showed significant increase in all biochemical parameters of HRS control except for ACE-1, CAT and creatinine clearance that experienced significant reduction along with dysbiosis compared to healthy control. Test groups showed improvement in all biochemical parameters with superiority to probiotic-green tea extract combination. Both treatments produced significant increase in fecal B. bifidum, S. thermophilus and L. bulgaricus and reduction of Staphylococci and Coliform. The effect of probiotic-green tea extract combination was more pronounced concerning the last three. Flavonoids and antioxidant activity of the extract were 1.325±0.01 mg quercetin/g and 98±1.66%, respectively. CONCLUSION: Administration of micro-encapsulated probiotic with or without alcohol green tea extract exerted significant prevention of HRS in rat with superiority to probiotic-green tea extract combination.

Application of liquisolid technology for promoting the renoprotective efficacy of walnut extracts in chronic renal failure rat model.

Chronic renal failure (CRF) is among the major health problems that could lead to increased morbidity and mortality among population. 'Nutraceuticals' is an emerging field for natural agents from plant foods that could reduce the progression of such disease. Many newly developed drugs are having bioavailability problems owing to their water insolubility. Liquisolid technique is one of the promising technological approaches to increase solubility and hence, drug absorption. The aim of the present research is to prepare and evaluate the renoprotective effect of the walnut extracts liquisolid formulations in CRF rat model. Saturation solubility study claimed PEG 400 and Tween 20 as good solubilizers for walnut extracts, thus chosen for preparation. The angle of slide was determined for the carrier; microcrystalline cellulose and coating material; silicon dioxide and liquid load factor was evaluated. Eight liquisolid systems were prepared employing 25% and 50% of liquid medication. Their flow and compressibility parameters showed good properties. Dissolution study was more in favor of formulations prepared using PEG 400. Of these, formulation F8 comprising carrier/coat ratio (10:1) and 50% liquid medication, showing superior dissolution properties was selected to perform stability and in-vivo evaluations. Two CRF induced rat groups received F8 at two oral doses (50 and 100 mg/kg). Biochemical and nutritional parameters were compared with both normal and CRF control rats. Results showed improvement of renal function, oxidative stress, antioxidant and inflammatory biomarkers as well as increased appetite and body weight gain on administration of both doses of walnut liquisolid formulation, F8.

Hepatic Regeneration and Reno-Protection by Fish oil, Nigella sativa Oil and Combined Fish Oil/Nigella sativa Volatiles in CCl4 Treated Rats.

The aim of the present research was to investigate the effect of fish oil, crude Nigella sative oil and combined fish oil/Nigella sative volatile oil as hepato-regenerative and renal protective supplements. The oils were administered as emulsions to rat model with liver injury induced by CCl4. Plasma activities of transaminases (AST and ALT) were evaluated as liver function indicators, while plasma creatinine and urea and creatinine clearance were determined as markers of kidney function. Plasma malondialdehyde (MDA), nitrite (NO) and tumor necrosis factor-α (TNF-α) were estimated to assess the exposure to oxidative stress and subsequent inflammation. Liver fat was extracted and their fatty acids´ methyl esters were determined using gas chromatography. Results showed that plasma activities of AST and ALT were significantly higher in CCl4 control group compared to control healthy group. Plasma levels of creatinine and urea increased significantly in CCl4 control, while creatinine clearance was reduced significantly in the same group. All rat treated groups given the three oil emulsions showed improvement in liver function pointing to the initiation of liver regeneration. The combination of fish oil/Nigella sative volatiles showed the most promising regenerative activity. Oxidative stress and inflammation which were increased significantly in CCl4 control group showed improvement on administration of the three different oil emulsions. Fatty acids methyl ester of liver fat revealed that rats treated with fish oil/Nigella sative volatile oil presented the highest content of unsaturated fatty acids (45.52% ± 0.81) while fish oil showed the highest saturated fatty acids (53.28% ± 1.68). Conclusion; Oral administration of oil emulsions of native fish oil, Nigella sative crude oil and combined fish oil/Nigella sative volatile oil reduced liver and kidney injury in rat model of CCl4 through exerting anti-inflammatory and antioxidant activity. Fish oil/Nigella sative volatile oil emulsion was the most promising hepato-regenerative and reno-protective formula among the different groups.

Prevention of renal dysfunction by nutraceuticals prepared from oil rich plant foods.

OBJECTIVE: To investigate the protective effect of extracts prepared from avocado, walnut, flaxseed and Eruca sativa seeds in a rat model of kidney dysfunction induced by intraperitoneal cisplatin. METHODS: Ethanol and petroleum ether extracts mixture was prepared from each plant. Six groups of rats were conducted; control healthy, cisplatin group and four test groups where rats were given daily oral dose of each extract mixture before cisplatin injection. Different biochemical and cytogenetic parameters and kidney histopathology were determined. Acute toxicity was tested for the nutraceuticals. Total phenolic contents, fatty acids (FA) and unsaponifiable matter were assessed in the extracts. RESULTS: Walnut ethanol extract showed the highest content of total phenolic. FA analysis revealed that all the studied plants were rich in unsaturated FA. Gas-liquid chromatographic investigation of the unsaponifiable matter showed the presence of campesterol, stigmasterol and ß-sitosterol in all the studied plants. Cisplatin treatment induced significant increase in plasma urea, creatinine and malondialdehyde along with significant reduction of plasma albumin, total protein, catalase and total antioxidant as well as reduction in creatinine clearance. Histopathological examination proved the induction of kidney dysfunction. Some sorts of chromosomal aberration and sperm-shape abnormalities were noticed after cisplatin treatment. Administration of extracts mixtures produced improvements in biochemical, histopathological and cytogenetic parameters. CONCLUSIONS: Administration of the studied nutraceuticals proved to possess protective role against cisplatin-induced nephrotoxicity, chromosomal aberration and abnormal sperms. All studied nutraceuticals showed complete safety.

Protective effect of clove oil and eugenol microemulsions on fatty liver and dyslipidemia as components of metabolic syndrome.

In the present research, the effect of clove essential oil (CO) and its major constituent, eugenol, formulated in water-based microemulsions, was studied on fatty liver and dyslipidemia in high-fructose-fed rats. Plasma and liver lipids, oxidative stress, inflammatory biomarker, and liver function were the assessed criteria. CO dispersed in water as conventional cloudy emulsion was also subjected to the same biological evaluations for comparison with the microemulsified form of this oil. Results showed that the particle size of CO microemulsion (COM) and eugenol microemulsion (EM) was 8.0 nm and 8.9 nm, respectively. Excess dilution and incubation of these microemulsions in 1.2 N HCl, that mimic stomach juice (without lipase), for 5 hours at 37 °C lead to the establishment of second population of larger particles with average diameter>100.0 nm. Biological evaluation revealed that rats of high fructose control group exhibited significant dyslipidemia, high plasma tumor necrosis factor-α, and elevated malondialdehyde. The same group of rats showed significant high liver total fat, triglycerides and cholesterol, and liver dysfunction compared to control normal rats fed balanced diet. Daily oral administration of CO conventional emulsion, COM, and EM produced significant improvement of all studied parameters. No significant change in all biochemical parameters was noticed when the groups given the different formulations were compared with each other. The study concluded that administration of CO conventional emulsion, COM, or EM produced significant improvement in fatty liver and dyslipidemia with consequent expected protection from cardiovascular diseases and other complications of fatty liver. Formulation of CO in microemulsion having particle size ∼ 8.0 nm did not enhance the protective effect compared with the same dose of CO dispersed in water as conventional macroemulsion, probably due to the ease of absorption of these bioactives in their native states. However, formulation in microemulsion provides a delivery system for oral administration of CO or eugenol in homogeneous, water-based, and thermodynamically stable dosage form during storage.

Epigenetic alterations by methylation of RASSF1A and DAPK1 promoter sequences in mammary carcinoma detected in extracellular tumor DNA.

Novel strategies for early detection of breast cancer, the most common and second most lethal cancer in women, are urgently needed. Silencing tumor suppressor genes via DNA methylation has established hypermethylation as one of the most frequent molecular alterations that may initiate and drive many types of human neoplasia including breast cancer. Detecting such epigenetic changes in DNA derived not only from tumor tissue, but also from bodily fluids, may be a promising target for the molecular analysis of cancer. In this study we examined serum, a readily accessible bodily fluid known to contain neoplastic DNA, from individuals with breast carcinoma. Using sensitive methylation-specific polymerase chain reaction, we searched for aberrant promoter hypermethylation of two normally nonmethylated genes: RAS association domain family member 1A (RASSF1A) and death-associated protein kinase 1 (DAPK1) in 26 patients with breast cancer, 16 patients with benign breast diseases, and 12 age-matched healthy controls. Hypermethylation of at least one gene was detected in 25/26 (96%) cancer patients, in 7/16 (43%) cases with benign breast diseases, and only 1/12 (8%) control subjects. Furthermore, methylation of both genes was found to be associated with ductal type of breast carcinoma. RASSF1A was hypermethylated in 18/26 cases (69%) and DAPK1 in 23/26 (88%). However, DAPK1 promoter methylation was more pronounced, as 12/23 DAPK1 methylated cases (52%) were strongly methylated (>75%) compared to the weaker methylation of RASSF1A (none of the cases with methylation at the level of >75%). These findings, if confirmed in studies of extended cohorts, may lead to useful clinical application in early diagnosis of breast cancer and better management of the neoplastic disease.