The effect of aqueous extract of Nepata glomerulosa Boiss. aerial parts on morphine withdrawal syndrome was investigated in mice. Dependence was induced by subcutaneous injections of morphine for 3 consecutive days. On day 4, morphine was injected 2 h prior to intraperitoneal injection of naloxone. The number of jumps during a 20 min. period after naloxone injection was considered as a measure of the withdrawal syndrome. Open field and rotarod tests were also performed. The results indicated that the aqueous extract (0.1 to 2.8 g/kg, s.c.) and diazepam (5 mg/kg) reduced the number of jumps (58±2.3 to 10±0.82, p<0.001 vs normal saline, 87±3.4). The extracts also reduced motion balance and locomotion activity. It is concluded that the aqueous extract of N. glomerulosa aerial parts could diminish morphine withdrawal syndrome.

Lung and Gastric cancer are one of the most common types of cancer in the world and also in Iran. For finding therapeutic ways, researchers have been focused on changes in carcinoma cells at molecular level. Some natural phytochemical compounds affect apoptosis path and play an important role in producing and regulating processes involved in cancer. In the present research, the toxic effect of hydro-alcoholic extract of Nepeta glomerulosa on A549 and AGS cell lines, and p53 expression changes have been evaluated. The plant shoots were collected from Binalud region from Khorasan Province and 80% ethanolic extract was prepared by maceration technique. The cytotoxic properties of extract on two cell lines in three times, i.e., 24, 48, and 72 hours were assessed by MTT assay. Apoptosis induction amount was evaluated by DAPI staining. p53 gene expression alterations were determined using Real-time PCR. According to the results, the IC50 was 15 and 7.5 mg/mL, for A549 and AGS respectively (P<0.05). The results of DAPI staining showed that under the influence of the extract at IC50 concentrations, apoptosis was induced in both strains compared to the control. The increasing effect of p53 gene expression on AGS was more that of reported for A549 cell line (P<0.05) that demonstrates the more antiproliferative effects of this species on gastric cancer cells through increasing p53 gene expression and apoptosis induction.

The current work aims to compare the effect of essential oil (pure and emulsified) Nepeta glomerulosa as natural preservatives for improving the shelf life of high-fat mayonnaise during 6 weeks of storage at 35 ºC. The nanoemulsion of Nepeta glomerulosa essential oil was prepared by the ultrasound cavitation method and physical properties were determined. Soy oil was substituted by 0.5 %w/w of the Nepeta essential oil or its nanoemulsion to compare the effect of natural and chemical (BHT and sodium-benzoate/potassium-sorbate) preservatives on the chemical (pH, acidity, peroxide-value, thiobarbituric acid-reactive-substances), microbial (acid-resistant bacteria, heterofermentative lactic acid bacteria, Escherichia coli, mold and yeast content, and salmonella) and sensory properties of mayonnaise. 1,8-Cineole (26.9%), α-Pinene (7.3%), and Limonene (6.8%) were the main compounds of this essential oil. The mean droplet diameter, PDI and viscosity of Nepeta nanoemulsions were 191.95 nm, 0.255, and 1.03 cP, respectively. The addition of the essential oil (pure or emulsified) significantly improved the oxidative stability of mayonnaise. There was no significant difference between the antioxidant activity of BHT and the essential oil. Over time, a decreasing and increasing trend was observed in pH and acidity, respectively, the slope of these trends was greater in the control sample. Nepeta essential oil and benzoate/sorbate significantly delayed microbial spoilage. The antimicrobial potential of the essential oil and its nanoemulsion were comparable with benzoate/sorbate activity (P > 0.05). At the end of storage, the overall-sensory scores of the sample containing the nanoemulsion were higher than the threshold-limit (>3) while in the 5th week, the control sample was introduced as unacceptable by evaluators. Nepeta nanoemulsion showed good potential to enhance the shelf-life of mayonnaise. Using essential oil in the form of nanoemulsion, not only reduced the consumption of essential oil but the bioactivity of the essential oil was protected for a longer time.

Introduction: The genus Nepeta with the common Persian name “ Pune-Sa” is one of the most important genera of Lamiaceae family. It has aromatic plants which are endemic of Iran. Its different species are distributed almost in all parts of Iran. Some of these species are valuable in medicine and used for medicinal purposes. Nepeta glomerulosa Boiss. is one of the most medicinal species of this family. This study was conducted to examine seed germination characteristics and to evaluate cardinal temperature of seed germination of N. glomerulosa. Materials and Methods: Seeds of N. glomerulosa were collected from Abade region of Fars province in the middle of September 2014. Treatments were including four levels of cold period (1, 2, 3 and 4 weeks at5º C) and four levels of potassium nitrate (0, 0. 1, 0. 2 and 0. 3 %). Factorial experiment with a completely randomized design in three replications was done, to evaluate seed germination percentage, germination rate, T50, mean germination time, root and shoot length, root and shoot fresh and dry weight. Also, the effects of 8 constant temperatures including 5, 10, 15, 20, 25, 30, 35 and 40 ˚ C and 3 variable temperatures including 5/15, 10/20 and 20/30 ˚ C (12h light/12h dark), was investigated to evaluate germination percentage, germination rate, mean germination time and cardinal temperatures of seed germination. Results and Discussion: The results showed that cold period and potassium nitrate interaction was significant only on root length. Potassium nitrate had no significant effect on germination rate, but the germination rate was increased by increasing the duration of being at5 ˚ C, so that the highest germination rate was observed in 3 and 4 weeks at5 ˚ C (1. 7 and 1. 2 /day respectively). The lowest germination rate (1. 0 /day) was observed in 2 weeks at5 ˚ C treatment. The maximum shoot length (51. 5 mm), shoot fresh weight (30 mg), shoot dry weight (1. 3 mg) and root dry weight (0. 6 mg) was obtained in 2 weeks at 5 ˚ C treatments and potassium nitrate 0. 1% was also the best treatment for abovementioned traits. Among the treatments, 3 weeks cold period and 0. 1 % potassium nitrate showed the most positive effect on seed germination characteristics of N. glomerulosa, although, had no significant effect on germination percentage. Based on the results of the second experiment, temperature had significant effect (P≤ 0. 01) on germination percentage, germination rate and mean germination time. The highest amount of germination percentage (76 and 77. 3 %), germination rate (4. 4 and 5 /day) and mean germination time (5. 1 and 5. 4 /day) were recorded in the range of 20 to 25 ˚ C. Germination rate and germination percentage were substantially reduced by increasing temperature above 25 ˚ C. The day and night variable temperatures had significant effect on seed germination percentage and germination rate of N. glomerulosa seeds. The highest amount of germination percentage (72 and 69. 3 %) and germination rate (4. 6 and 5. 7 /day) were observed at10/20 and 20/30 ˚ C. The results of the relationships between temperature and germination rate showed that base, optimum and maximum temperatures for seeds of N. glomerulosa were 15 ˚ C, 22 ˚ C and 46 ˚ C, respectively. Conclusion: The overall results showed that most chilling occurred under natural conditions during winter and early spring which caused increasing in seed germination of lots of plants. This also found that increasing in chilling period significantly improved the seed germination characteristics of N. glomerulosa. Also, treating with potassium nitrate had significant effects on germination properties of N. glomerulosa. Although applying potassium nitrate and cold period did not show interaction effects on seed germination. The effects of different day and night temperatures on seed germination rate and percentage have been reported in many medicinal and aromatic plants as may be due to the similarity of these conditions to the natural conditions. Comparing cardinal temperatures of N. glomerulosa with the other species of this genus indicated that they have different cardinal temperatures. Many of researchers reported that germination cardinal temperatures are significantly different among species and different genotypes of species. Determining of cardinal temperatures helps us to estimate temperature range of wild species in natural areas, planting time and also suitable areas for cultivation.

Nepeta glomerulosa Boiss. is a medicinal plant used in traditional medicine. The aim of this study was to evaluate the anti-melanogenesis inhibitory activity of Methanol (MeOH), n-hexane, acetate (EtAc), dichloromethane (CH2Cl2), n-butanol (BuOH), and H2O extracts isolated from N. glomerulosa in B16 melanoma cell line. The B16F10 cell line viability after treatment with increasing concentrations of different extracts of the plant (50-200 μ g/ml) was measured by using MTT. The inhibitory effect on synthesis of melanin, mushroom tyrosinase activity, cellular tyrosinase, and effect on oxidative stress was determined by the colorimetric and fluorometric method. The data showed that at concentrations <200 μ g/ml, all extracts did not show significant toxicity on melanoma cells. The amount of melanin synthesis by MeOH and CH2Cl2 extracts and mushroom tyrosinase activity by the MeOH extract declined in B16F10 cells. In addition to the capacity of MeOH, n-hexane and CH2Cl2 extracts in decreasing the amount of reactive oxygen species (ROS) in melanoma cells all extracts revealed the remarkable antioxidant activity. Our results showed the melanogenesis inhibitory and antioxidant effects of N. glomerulosa on B16F10 cells may recommend a novel agent in diminishing skin pigmentation and skin aging in cosmetic industry.

To investigate the cardinal temperature of three medicinal species of Nepata haussknechtii, N. pogonosperma and N. glomerulosa subsp. staffina and determining the appropriate temperature and time conditions for their planting, a factorial experiment was carried out in the form of a completely randomized design in three replications in 2023 in the Seed Technology Research Laboratory of the Gene Bank of Natural Resources of Iran. Three non-linear regression models, including dent-like, segmented and beta, were evaluated at eight temperature levels (5, 10, 15, 20, 25, 30, 35 and 40°C) to describe the response of the seed germination rate of three Nepeta species to temperature. Statistical indices such as root mean square (RMSE) and coefficient of explanation (R2) were used to compare the models.The results showed that the parameters estimated using the dent-like model in N. haussknechtii include base temperature, optimal lower temperature, optimal upper temperature, ceiling temperature, R2 and RMSE values were 9.01°C, 20°C, 25°C, 36.21°C, 0.94 and 0.11, respectively; segmented model in the N. pogonosperma include base temperature, optimal temperature, ceiling temperature, R2 and RMSE values were 9°C, 23°C, 36.14°C, 0.95 and 0.31, respectively; and beta model in the N. glomerulosa subsp. staffina include base temperature, optimal temperature, ceiling temperature, R2 and RMSE values were 9.84°C, 24.95°C, 37°C, 0.99 and 0.03, respectively had more confidence. The results of this experiment showed that in the different studied species of Nepeta, due to the similar initial growth rate, they have the same germination peak.