ANTICANCER ACTIVITY OF SEMEN PROTEIN NANO SUSPENSION IN MICE MODEL

 

ANTICANCER ACTIVITY

Semen is a complex substance created by the male reproductive organs. The fluid is made mostly of water, plasma, and mucus (a lubricating substance). It also contains 5 to 25 calories and is made up of small amounts of essential nutrients including:

Citrate

Protiens

Zinc

Calcium

Magnesium

Potassium

AntiCancerActivity

Animals 

A total of 60, 6–7-week-old, Swiss albino mice (weight range, 25–30 g) of both genders (Samtako Bio Korea, Osan, Korea), were divided into six groups, each consisting of 12 animals, and were used to assess biological activity. Animals were maintained in an air-conditioned room at a temperature of 22±1°C and a humidity of 55±1% with a 12 h light/dark cycle. They were fed a standard commercial rodent pellet diet (Samtako Bio Korea), and had ad libitum access to water. The animals were allowed to acclimate to the environment for seven days prior to the experimental session. All animal experiments were performed in accordance with the guidelines of the Institutional Animal Ethics Committee, Atish Dipankar University of Science & Technology (Dhaka, Bangladesh). Animal treatment and maintenance for acute toxicity and effects were strictly followed in accordance with the Principle of Laboratory Animal Care and the Animal Care and Use Guidelines of Atish Dipankar University of Science & Technology

Acute toxicity study 

A total of 30, 6–7-week-old, Swiss albino mice (weight range, 25–30 g) of both genders (Samtako Bio Korea), were divided into five groups (n=6), and were used to assess acute toxicity activity. The test was performed using increasing oral doses of SPNS in distilled water (50, 100, 200, 500 and 1,000 mg/kg body weight) that were administered orally at 20 ml/kg to each test group. The normal control group received distilled water (1 ml). Following treatment, mice were allowed to feed ad libitum and observed for 48 h for any mortality or behavioral changes.

SPNS - SEMEN PROTEIN NANO SUSPENSION

Determination of tumor and packed cell volume (PVC) 

Mice were dissected and ascetic fluid was collected from the peritoneal cavity. Volume was determined using a graduated conical centrifuge tube, following which the PCV was determined by centrifuging the fluid at 1,000 × g at 4°C for 5 min.

Viable and nonviable cancer cell count 

Ascetic fluid was collected in a white blood cell (WBC) pipette and diluted 100 times using PBS. A drop of the diluted suspension was subsequently placed on a Neubauer counting chamber, and the cells were stained with trypan blue (0.4% in normal saline). Cells that did not take up the dye were considered viable, whereas those that did were considered non-viable. These viable and non-viable cells were counted using the following equation:

Cell count = (number of cells × dilution factor)/(area × thickness of liquid film)

Determination of median survival time and percentage increase in lifespan 

Mortality was monitored by recording the percentage increase in lifespan (% ILS) and the median survival time (MST)

Estimation of hematological parameters 

Collected blood was used to estimate the hemoglobin (Hb) content, red blood cell (RBC) and WBC count. Differential counts of WBC were performed using Leishman stained blood smears

Treatment schedule 

Mice were divided into six groups (n=12) and provided with food and water ad libitum. All animals in each group received EAC cells (2×106 cells/mouse, i.p.) except group I, which served as the normal saline control (5 ml/kg body weight i.p.). Group II served as the EAC control. Following a 24-h period post-EAC transplantation, group VI received bleomycin 0.3 (i.p.) mg/kg body weight (positive control), and groups III, IV and V were treated with 50, 100 and 200 mg/kg body weight (p.o.) semen protein nano suspension (SPNS) respectively, for nine consecutive days. Following 24 h from the last dose, the animals were fasted for 18 h; six animals in each group were subsequently sacrificed by cardiac puncture to estimate hematological parameters, as well as to determine anticancer activity. The remaining mice were provided with food and water ad libitum and observed to determine if there were any changes in lifespan. The anticancer activity of the extract was evaluated in Semen Protiens nano suspension (SPNS) animals.

Statistical analysis 

All values are expressed as the mean ± standard error of the mean of three replicated experiments. The analysis was performed using SPSS statistical package for WINDOWS (version 16.0; SPSS, Inc., Chicago, IL, USA). Results associated with the reducing power activities were statistically analyzed by applying the Student's t-test, and P<0.001 was considered to indicate a statistically significant difference. All in vivo data were subjected to analysis of variance followed by Dunnett's test, and P<0.05 was considered to indicate a statistically significant difference

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