动物实验,草本洁面乳 z-bo 2020-09-03 460

　　Background: A healthy facial cleanser has an herbal formula. The cleanser contains a variety of medicinal plant extracts with androgen, antipyretic, analgesic and anti-inflammatory potential. We are using male Wistar rats to evaluate its safety and warn that BSC has been widely used and has not been evaluated by the US Food and Drug Administration (NAFDAC). Following standard toxicity guidelines, a single BSC capsule (total 442 mg) was dissolved in distilled water (80 mg/ml) and administered to rats by gavage. Experimental design: Oral acute toxicity study: 30 male Wistar mice (average 20 g) were housed in captivity, randomly divided into 6 groups, and adapted for 2 weeks before administration. The oral BSC concentration is 250, 500, 1000, 2000 and 4000 mg/kg, and animals are allowed to eat freely. The control group took 0.1ml distilled water orally. Observe the mortality and behavioral changes of the mice within 24 hours after administration (hyperactivity, circulatory movement, tilted hind limbs, increased eating habits, scraping of the jaw, etc.). All animals were further monitored within 14 days after administration. Acute abdominal toxicity study: 30 male mice (average 20 g) were placed in different cages and randomly divided into 5 groups. BSC doses of 200, 400, 800 and 1600 mg/kg were given intraperitoneally. The control group took 0.1ml distilled water orally. Observe the mortality, behavioral changes and toxicity of the mice within 24 hours after administration. Animals with a BSC of 200 mg/kg showed restlessness, climbing, bulimia, tilting of hind limbs, and circular movement. Two hours later, animals given doses higher than 400 mg/kg showed signs of restlessness, weakness, flat abdomen and tearing. It was observed that weight loss occurred within approximately 6-10 hours, and most animals shivered. Observe and record the mortality of each group of animals 24 hours after administration. 14 days after administration, all animals were further monitored. The median lethal dose was estimated according to the Finney method. Subchronic toxicity study: 32 adult male Wistar rats weighing 150-300 grams were divided into 4 groups, each with 8 rats. The first group, the control group, drank 10 ml of distilled water every day. The second, third and fourth groups received 250 mg/kg, 500 mg/kg and 1000 mg/kg BSC, respectively. During the experiment, the rats were weighed weekly. Pay attention to changes in animal behavior. Collect blood samples and tissues for clinical testing: rats in the experimental group were given the drug for 60 days. Collect blood samples within 24 hours after the last dose and place them in anticoagulated and non-anticoagulated test tubes. Remove the neck and put the animal to death. The blood sample of the anticoagulant was centrifuged at 4200pm for 5 minutes to separate the serum for biochemical analysis. The liver, kidney, heart and brain were dissected, weighed and homogenized with 4 times the volume of buffer (0.1M, ph7.4). Some organs will be taken out for histological studies, while other organs will be homogenized for biochemical analysis.

　　Biochemical analysis: liver and kidney function test: evaluate liver function by measuring serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP). Serum creatinine (CREA) and blood urea nitrogen (BUN) levels were measured to assess renal function. Protein and uric acid: In order to evaluate the synthetic function of the liver, it is necessary to measure the levels of total serum protein (TP) and albumin (ALB), as well as the level of uric acid.

　　Blood lipids: Use the kit method to determine serum total cholesterol (TC) and triglycerides (TG). And measure the high density lipoprotein (HDL) and low density lipoprotein (LDL) in the serum. Antioxidant and oxidative stress: Measure glutathione (GSH) levels and glutathione-S-transferase (GST) activity. The activity of superoxide dismutase (SOD) and catalase were measured. Lipid peroxidation is determined by measuring the thiobarbituric acid reactant formed. Hematology evaluation: An automated hematology analyzer is used to measure red blood cells (PCV), hemoglobin (Hb), platelets (PLT) and total white blood cells (TWBC).

　　Electrolytes: the content of sodium ion (Na +), potassium ion (K +), chloride ion (Cl-), total calcium ion (tca2 +) and intracellular calcium ion (calcium) in the flame photometer. I measured histological evaluation : Collect liver, kidney or heart tissue samples, and conduct histological examination by methods such as fixation, dehydration, transparency, penetration, embedding, sectioning and staining. In order to ensure a good fixation, the tissue is trimmed to a thickness of approximately 5 mm to achieve a good fixation. Fix the tissue with 10% formaldehyde solution and transfer it to 50% (70%, 80%, 85%, 95, 100%) ethanol for 2 hours. The paraffin tissue was then placed on a wooden block and arranged in order of size. The rotary microtome is used for continuous sectioning, with a thickness of 10μm. Eight sections were obtained from each treated organ of each animal. After staining, DPX was used as a fixative and subjected to microscopic examination. Results: Acute oral toxicity test: Table 1 shows the results of the acute oral toxicity test of BSC at doses of 250, 500, 1000, 2000, and 4000 mg/kg. The animals did not die at the given dose. Compared with the control group, no changes in toxicity were observed at a dose of 250 mg/kg, but at a dose above 500 mg/kg, motor activity, mild scratching, and mild scratching within 2 hours after administration were reduced . I saw scratches. Overactivity and weakness. Acute abdominal toxicity test: Table 2 shows the acute abdominal toxicity test. We observed changes in mortality or toxicity in mice treated with BSC200, 400, 800, and 1600 mg/kg. At 200 mg/kg, the animal exhibited restlessness, climbing, bulimia, hindlimb tilting and circular movement. Two hours after administration, rats at a dose of 400 mg/kg showed signs of restlessness, weakness, flat abdomen, tearing, and shallow breathing. Most animals are shaking for about 6-10 hours. The average lethal dose (LD50) is 600 mg/kg. Liver function: Table 3 shows that compared with the control group, the serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) of the treatment group did not change significantly. (P\→0.05). 250 mg/kg BSC reduced ALT and ALP levels, respectively. When taking 250 mg/kg (32.4%), 500 mg/kg (39.2%) and 1000 mg/kg (51.4%), the dose-dependent increase in AST levels was statistically significant (P\0.05). At 500 mg/kg (41.4%) and 1000 mg/kg (29.3%) doses, ALT levels increased significantly (P\→ 0.05). Blood lipids: Table 4 shows a slight decrease in serum TG levels, down 11.6%, 9.8% and 1.5% respectively. However, the lowest dose of 250 mg/kg may slightly increase serum TC levels. At 250 mg/kg and 500 mg/kg doses, serum low-density lipoprotein (LDL) decreased by 14.7% and 4.6%, respectively, while the 1000 mg/kg dose increased LDL slightly by 28.5%. Renal function, protein synthesis and uric acid: After the administration of 500 mg/kg and 1000 mg/kg, blood urea nitrogen levels increased by 11.8% (P \u003c0.05) and 13.7% (P \u003c0.05). The creatinine level increased by 7.1% after taking 250 mg/kg. However, serum albumin (ALB), total protein (TP) and UA did not change significantly. After administration of 500 mg/kg and 1000 mg/kg, the levels of ALB, TP and UA increased. Hematology evaluation: After taking 250mg/kg (P \u003cu\→ 0.05) and 500mg/kg (P \u003cu003c0.05), the number of red blood cells can be increased. However, compared with the control group, the hemoglobin level did not change significantly. 250 mg/kg can increase the white blood cell count by 62.6%, while 500 mg/kg can increase the platelet count by 50.6%. Electrolytes: 250mg/kg orally? 1000mg/kg will not cause a significant increase in sodium ions. The calcium content decreased at 250 mg/kg, while the calcium content increased at medium and high doses. Serum potassium, chloride and intracellular calcium levels remained unchanged. Antioxidant in tissue: Antioxidant in liver: Under low dose (250mg/kg) and high dose (1000mg/kg), liver GSH content increased to 68% and 49.2%, respectively. Compared with the control group, there was no significant difference between GST and superoxide dismutase. However, 1000 mg/kg can increase liver CAT levels by 54.1%. Kidney antioxidants: The highest dose (1000mg/kg) can increase renal GSH levels. Renal GST levels decreased at all doses. Only the lowest dose (250 mg/kg) was shown to increase CAT levels. 250mg/kg and 500mg/kg can increase kidney SOD levels. Cardiac antioxidants: At the highest dose (1000mg/kg), glutathione levels increased by 9.7% and cardiac GST levels decreased by 54%. 500mg/kg and 1000mg/kg can increase CAT levels in the heart. The SOD level of the rat heart was significantly reduced by 30.1% (250 mg/kg), 40.5% (500 mg/kg) and 39.1% (1000 mg/kg). Brain antioxidants: The dose-dependent increase in GSH levels in brain tissue was 18.8%, 96.4% and 212.3%, respectively. 250 mg/kg reduces the level of GST in the brain. 500mg/kg and 1000mg/kg can increase the level of CAT in the brain. The brain SOD level of all treatment groups decreased.

　　Histological evaluation: control group (distilled water, 10ml/kg): control group (distilled water, 10ml/kg, A) and BSC (250mg/kg, B, 500mg/kg, C) normal heart has been indicated. However, BSC (1000mg/kg, D) does show cardiac inflammation. BSC (250mg/kg): The control group (distilled water, 10ml/kg, E) and BSC (250mg/kg, F; 500mg/kg, G; 1000mg/kg, H) showed normal kidneys. BSC (500mg/kg): The control group (distilled water, 10ml/kg, I) and BSC (250mg/kg, J, 500mg/kg, K) showed normal liver. However, BSC (1000 mg/kg, L) shows congestion of the liver sinusoids and central veins.

　　Conclusion: Using Wistar rat grade, BSC is considered relatively safe. Due to mild changes in the liver and heart, the herbal BSC is considered to be used with caution.