Effect of pomegranate peel polyphenols on mTOR/HIF-1α/IL-17 signaling pathway in a rat auriclular model of acne / 中华皮肤科杂志

Objective:To explore the anti-inflammatory effect of pomegranate peel polyphenols on a rat auriclular model of acne and its mechanism of action.Methods:Totally, 36 specific-pathogen-free SD rats were randomly divided into 6 groups: blank group, model group, low-, medium- and high-dose pomegranate peel polyphenol groups and positive control group. In all groups except the blank group, 0.5 ml of 100% oleic acid was applied to the openings of bilateral auricular ducts once a day for 3 consecutive weeks, followed by subcutaneous injections of 50 μl of Propionibacterium acnes suspension at the oleic acid-applied sites once a day for 3 consecutive days, so as to establish a rat auriclular model of acne. After the model was confirmed to be successfully established by naked eyes, the low-, medium-, high-dose pomegranate peel polyphenol groups were topically treated with 0.5 mg of 1.4%, 2.8%, 5.6% (mass fraction) pomegranate peel polyphenol ointment respectively, the positive control group was topically treated with 0.5 mg of clindamycin hydrochloride gel, and the blank group and model group were topically treated with the same amount of distilled water. All the topical treatments were performed twice a day for 2 consecutive weeks. Twenty-four hours after the last topical treatment, abdominal aortic blood samples were collected, and enzyme-linked immunosorbent assay (ELISA) was conducted to detect the serum level of interleukin 17 (IL-17) in rats; rat auricular tissues were resected, hematoxylin-eosin (HE) staining was performed to observe histopathological changes of the skin tissues in each group, and immunohistochemical study to determine the expression of mammalian target of rapamycin (mTOR) , hypoxia-inducible factor-1α (HIF-1α) , and retinoic acid-related orphan receptor-γt (RORγt) in local tissues. Data meeting the assumptions of homogeneity of variances were analyzed by using one-way analysis of variance, and those that did not meet the assumptions of homogeneity of variances were analyzed by using Kruskal-Wallis H test; multiple comparisons were performed by using least significant difference- t test. Results:Compared with the model group, the pomegranate peel polyphenol groups and positive control group showed marked improvement in cysts, desquamation, crusts and epidermal keratinization, and reduced infiltration with inflammatory factors in the dermis at the modeling site. The serum level of IL-17 was significantly lower in the low-, medium- and high-dose pomegranate peel polyphenol groups (61.03 ± 5.99 ng/L, 55.35 ± 2.24 ng/L, 54.35 ± 4.29 ng/L, respectively) , positive control group (48.11 ± 4.07 ng/L) and blank group (42.10 ± 5.62 ng/L) than in the model group (70.24 ± 3.30 ng/L; t = 3.12, 5.34, 5.70, 8.29, 10.54, respectively, all P<0.05) . Immunohistochemical study revealed that the HIF-1α expression level was significantly lower in the low-, medium- and high-dose pomegranate peel polyphenol groups (0.29 ± 0.05, 0.29 ± 0.03, 0.33 ± 0.02, respectively) and positive control group (0.30 ± 0.01) than in the model group (0.41 ± 0.04; t = 4.89, 5.50, 3.62, 5.21, respectively, all P<0.05) ; the RORγt expression level was significantly lower in the low- and high-dose pomegranate peel polyphenol groups (0.28 ± 0.02, 0.31 ± 0.04, respectively) than in the model group (0.35 ± 0.02, t = 3.68, 2.18, respectively, both P<0.05) ; there was no significant difference in the mTOR expression level among these groups ( P = 0.119) . Conclusion:Pomegranate peel polyphenols could improve inflammatory reactions in the rat auriclular model of acne, which may be related to the down-regulation of HIF-1α/RORγt signaling pathway.

Effect of pomegranate peel polyphenols on sebaceous gland spots and AKT/Sox9 signaling pathway in golden hamsters / 中华皮肤科杂志

Objective:To evaluate the effect of pomegranate peel polyphenols on sebum secretion in golden hamsters, and to explore its possible mechanisms.Methods:Thirty golden hamsters were randomly and equally divided into 5 groups: (ointment) vehicle group, 0.48%-, 0.96%-, 1.92%-pomegranate peel polyphenol ointment groups, and retinoic acid cream group. Corresponding cream or ointments were applied to bilateral sebaceous gland spots of the golden hamsters at a dose of 1 gram twice a day for 4 consecutive weeks. The area of bilateral sebaceous gland spots was measured on days 0, 7, 14, 21 and 28 after the start of treatment, which was calculated by the maximum longitudinal diameter multiplied by the maximum transverse diameter. Twenty-four hours after the last treatment, immunohistochemical study was conducted to determine the expression of AKT/Sox9 signaling pathway in sebaceous gland spots resected from the golden hamsters. The area of sebaceous gland spots in these groups at different time points was compared by repeated measures analysis of variance, and other data were analyzed by one-way analysis of variance or Kruskal-Wallis H test. Results:The area of sebaceous gland spots was significantly smaller in the 0.96%-, 1.92%-pomegranate peel polyphenol ointment groups (50.48±2.41 mm 2, 48.24±2.56 mm 2, respectively) and retinoic acid cream group (48.31±2.76 mm 2) than in the vehicle group (57.99±3.29 mm 2; t=2.69, 3.98, 3.65, P=0.012, 0.001, 0.001, respectively) . Sox9 expression was significantly lower in the 1.92%-pomegranate peel polyphenol ointment group (0.39±0.04) and retinoic acid cream group (0.38±0.03) than in the vehicle group (0.44±0.02, P=0.040) . However, there was no significant difference in AKT expression among the 5 groups ( F=1.645, P=0.199) . Conclusion:Pomegranate peel polyphenols can reduce the sebaceous gland spot area and inhibit sebum secretion in golden hamsters, which may be related to the inhibition of Sox9 expression.

Analysis of Urinary Metabonomics Pathway in Rats with Chronic Glomerulonephritis Intervened by Tannins from Pericarpium Granati / 中国药师

Objective: To analyze the metabolicomics pathway in the rats with chronic glomerulonephritis intervened by tannins from Pericarpium Granati.Methods: Signal pathway analysis was carried out using the KEGG database and molecular metabolite annotation was performed using HMDB database.The enzyme or transporter and its related properties were analyzed.The metabolite path visualization was carried out by using MetPA network software.Results: The analysis of biological metabolism pathway showed that 12 metabolites involved in 16 metabolic pathways.The pathway of tryptophan metabolism, citrate cycle, phenylalanine metabolism, cysteine and methionine metabolism showed notable changes (P<0.05).Conclusion: The changes of phenylalanine metabolism, citrate cycle, tryptophan metabolism, cysteine and methionine metabolism in the rats with chronic glomerulonephritis intervened by tannins from Pericarpium Granati participate the pathological process.

Effect of Tannins from Pericarpium Granati on Hepatic Microsomal Enzyme in Rats / 中国药师

Objective:To study the effect of tannins from Pericarpium Granati on hepatic microsomal enzyme in rats. Methods:Thirty Wistar rats were randomly divided into five groups:the blank control group, high, medium and low dose groups of tannins from Pericarpium Granati, the positive control group of phenobarbital sodium. The blank control group was given physiological saline. The three different doses groups were respectively given tannins from Pericarpium Granati orally at the dose of 150,100 and 75 mg·kg-1 · d-1 for 7 days. The positive control group was given phenobarbital sodium 80 mg·kg-1 ·d-1 with intramuscular injection for 5 days. At the end of the experiment, the activity ofⅠandⅡphase metabolic enzymes in liver microsomes of each group was determined by UV. Results:Compared with the blank control group, the high, medium and low dose groups of tannins from Pericarpium Granati could sig-nificantly decrease the content of CYP 450 and CYPb5, and inhabit the activity of ADM (P<0. 01);the high and medium dose group could significantly inhibit ERD enzyme activity (P<0. 01);the high dose group could significantly reduce GST enzyme activity (P<0. 01). Conclusion:Tannins from Pericarpium Granati has notably inhibitory effect on hepatic microsomal enzyme in rats, which can reduce the expression of CYP3A and CYP2E1 in a dose-dependent manner.

ß-Cyclodextrin anchoring onto pericarpium granati-derived magnetic mesoporous carbon for selective capture of lopid in human serum and pharmaceutical wastewater samples.

Functionalized magnetic carbonaceous nanomaterials, which are important materials with many practical and research applications in biomedical, pharmaceutical and biological fields, have recently attracted much attention. In this study, a magnetic mesoporous carbon coated with ß-cyclodextrin (MMC@ß-CD) was synthesized for the first time from natural pericarpium granati (PG). The as-obtained MMC@ß-CD has high surface areas (203 m(2)g(-1)), large pore volumes (0.16 cm(3)g(-1)), relatively broad mesoporous sizes (6.8 nm) and a high saturation magnetization of 26.2 emu g(-1), which is sufficient for magnetic separation by an external magnetic field. The MMC@ß-CD was used as an innovative adsorbent for magnetic solid-phase extraction of lopid via host-guest interaction prior to spectrofluorometric analysis. The proposed method was successfully applied to analyze lopid in human serum and pharmaceutical wastewater samples with recoveries in the range of 85.0-103.5% for the spiked samples. Overall, this work not only provides an inexpensive and eco-friendly method to fabricate MMC@ß-CD (or MMC) from PG, but also develops a highly selective approach for capture of lopid in biological samples and environmental substances.

Polyethylene glycol as a novel solvent for extraction of crude polysaccharides from Pericarpium granati.

Polyethylene glycol aqueous solution (PEGs), an environmentally friendly solvent was used, for the first time, to develop an extraction method for crude polysaccharides from pericarpium granati with ultrasonic-microwave-assisted extraction (UMAE). Compared with other extraction solvents, PEGs, especially PEG400, displayed a higher extraction yield. The extraction conditions were an ultrasonic power of 240 W, a microwave power of 365 W, a PEG400 concentration of 30% and a ratio of liquid to raw material of 20 mL/g, optimized by using Box-Behnken experimental design. Under these conditions, an extraction yield of 7.94 ± 0.3% (n=3) was obtained, which is about 25% higher than that with water as a solvent, and the polysaccharides content in pericarpium granati was 6.56 ± 0.01 mg/g expressed as glucose.