Anti-Tumorigenic Effects of Sage in Female Rate

Anti-Tumorigenic Effects of Salvia Officinalis Regarding the
important role of protease enzymes in various complications...

which is, in fact, an in-vitro carcinogen [4]. Numerous studies have reported 20 mg/ml DMBA can properly induce cancer [5]. DMBA was fed to animals three times a week for one month at the dosage of 10 mg/ml.

Garden Sage Preparation
Salvia officinalis seeds were purchased from Maxima Company.
Following cultivation in the greenhouse, its aerial parts including leaves were utilized. The plant extract was prepared using liquid nitrogen and delivered to medical sciences animal room in as a Hydro-alcoholic solution.
In this method, tannic acid was used as a standard. After plotting the calibration curve and determining the linear equation, the total phenolic content amount was expressed as mg tannic acid per liter of sample. To assess the anti-oxidant characteristic of the essence using DPPH strategy (the method developed by Ebrahim Abadi et al. [7]), beta-carotene/linoleic acid bleaching [8] and reduction potential [9] methods were implemented. The anti-oxidant activity was represented as IC 50 which indicates the required concentration of the essence to inhibit 50% of DPPH free radicals.
In the case of beta-carotene methodology, IC50 refers to the essence concentration capable of 50% prevention of beta-carotene/linoleic acid emulsion bleaching. Concerning the reduction potential, IC50 can be defined as the sample concentration at which the absorbance equals to 5.0. The AOCS standard method [10] was also employed to determine the amount of peroxide in oil samples packed by HPMC films.

Laboratory Rats
Female Wistar rats were acquired from Bushehr Medical Sciences Animal Department. Animals were kept in the animal room under controlled conditions involving 16:8 light and darkness cycles and temperature of 25℃.

Animal Classification
54 mature female rats were divided into 4 groups. The control group consisted of 8 rats, which received 3 ml of sunflower oil in two hours, every two days, for three consecutive weeks. The case group included 20 cancerous rats which orally received aqueous garden sage extract, every day. Rats were treated for 6 months and analyzed regarding their cancer stage (creation and progression).
The treatment group also encompassed 20 cancerous rats which only received distilled water on a daily basis.

Statistical Analysis
All the experiments were performed with three replications.
The obtained data were reported as mean plus standard deviation.

Results
The results were analyzed by SAS-2000 software (edition 9) using a factorial experiment based on a completely randomized design, and the least significant difference (LSD) mean comparison method at the probability level of 95%.

Findings Total Phenolic Content and Anti-oxidant Activity
Garden sage essence encompassed the total phenolic content of 5.276 milligrams of tannic acid per liter of sample. Its antioxidant activity is displayed in Table 1 based on IC 50 , through the implementation of DPPH free radical inhibition method, beta-carotene/linoleic acid emulsion bleaching prevention, and reduction potential, all of which were spectrophotometrically measured. Lower IC 50 indicates higher antioxidant activity of the essence.
In all the models, the lowest IC 50 belonged to BHT. Here, the necessary amount of garden sage to cause 50% inhibitory effect is higher in comparison with that of BHT, which implies higher BHT potential (p<0.005).

Study of DMBA-Induced Cancer Groups Receiving Garden Sage Aqueous Extract
Breast tissue changes in cancer-bearing rats were compared in both groups (treated with distilled water and garden sage aqueous extract). Lobules were counted using light microscopy T-test statistical analysis was also exploited to evaluate the changes. The results indicated a reduction in the number of lobules in the group receiving garden sage aqueous extract, and this reduction was statistically significant in the 4 th and 6 th months.

Total Phenolic Content and Antioxidant Activity
Total phenolic content is an indicator of the antioxidant capacity of the plant. Garden sage possesses higher phenolic content in comparison with many other medicinal herbs including Chamomile, Pot marigold, Rhubarb, and Lavender [11]. Garden sage essence also contains ~60% oxygen-bearing monotropins and about 20% oxygen-bearing hydrocarbons. Among the most prominent compounds of these groups, alpha-thujene, camphor, viridiflorol, borneol, 1,8-cineol, beta-thujene, and bornyl acetate can be mentioned whose contents vary depending on the circumstances.
Therefore, these groups may exhibit distinct antioxidant potential in different conditions [12]. Viado-Martos et al. [6] reported the total phenolic content of garden sage essence as 98.122 mg galic acid per liter of sample. Estrogenic compounds initiate the breast growth and lactation system and account for breast development in adult females. The phytoestrogens are structurally and functionally similar to 17-beta-sterol. They include several chemical groups among which isoflavonoid can be mentioned [13]. Phytoestrogens display estrogen-compatible effects at low blood estrogen concentrations. In contrast, they behave oppositely at high estrogen levels [14].
Phytoestrogen estrogenic activity is concentration-dependent [15]. At low concentrations, phytoestrogens induce growth and reproduction of breast tissues similar to estrogens, while at high concentrations, they prevent excessive growth and proliferation of breast tissues and DNA synthesis [16]. Garden sage contains genitein phytoestrogen [17]. Findings demonstrated that DMBA impact on breast tissue is similar to that of estrogen, and it increased the number of lobules in rat's breast tissue. However, rats treated by garden sage aqueous extract did not exhibit similar results. Garden sage extract led to the reduction of lobule counts in the breast tissue of DMBA-induced cancerous rats when compared with the group treated by DMBA and water. Such an effective therapeutic impact can be attributed to phytoestrogens present in the garden sage herb. DPPH: 2,2-diphenyl-1-picrylhydrazyl used to assess the radical scavenging potential of the substances. It is a dark-colored crystalline powder composed of stable free-radical molecules.

Acknowledgement
The author wish to thank Mrs zarei lab tecnition of research beesat center of shiraz,for running the GC/MS profiles.

Declaration
Availability of data and materials: "Data sharing not applicable to this article as no datasets were generated or analyzed during the current study.