The Preclinical Preliminary Safety Evaluation On The Effective Constituent Of Mongolian Medicine Compound Terunshudule

Based on previous research in vitro and vivo on the antibacterial, anti-inflammatory and immune functions on cow mastitis of the active ingredients in Mongolian medication-Terunshudule compound, we developed acute, sub-acute and chronic toxicological experiments in vitro and vivo to evaluate its safety thus ensuring its nontoxicity to cows. We also examined the antimicrobial spectrum to provide evidence for clinical application.The results of our experiments were as follows:1.Acute toxicity test:maximum tolerance method was adopted by setting up four dose groups through single administration and the observation period was 7days. The maximum tolerate dose of the active ingredients in Terunshudule was 5000mg/kg b.w, which was the basis of the subsequent safety dose tests.2.Sub chronic toxicity test:the aim was to evaluate accumulative toxicity, tolerance and toxicity on key target organs of long-term administration. The groups were high-and low-dose groups and negative controls. Thirty-day feeding test. At the end of the entire administration, the spirits of rats were normal, and the general trend of their food intake and weights was increasing, which showed no statistical significant difference compared to controls (P>0.05). Viscera index was also normal. Although there are statistical differences in terms of blood routine examination and blood biochemical index, the fluctuations are all within the normal range. At the end of drug administration, no macroscopic lesions were found in the rat autopsy, and no obvious degeneration necrosis and hemorrhage were observed in the biopsy under a microscope, indicating that the drug was nontoxic to organs. The results proved that long-term drug administration presented no accumulative toxicity and had no damage to organs.3.Mutagenic test:we did sperm malformation test on mice by setting up high- and low-dose groups, and positive and negative controls. The drug was administered for a continuous 5 days. Mice were executed and their testicles were separated on the 35th day since drug administration. We observed morphology of sperm under a microscope. The results showed statistically significant difference between positive and negative controls (P<0.05). When compared to negative controls, there were no significant differences in each dose group (P>0.05). However, there were significant differences between each dose group and positive controls (P<0.05). The results indicated that the active ingredients in Terunshudule compound did not cause sperm deformity in mice, therefore presenting no damage to mice reproductive cells.4.Micronucleus test of bone marrow cells in mice:the groups were high-and low-dose groups, positive and negative controls. The drug was administered for a continuous 5 days, and mice were executed 6 hours after last administration. Bone marrow cells were observed under a microscope. The results showed that there was no significant difference between each dose group and negative control group (P>0.05). However, there was significant difference between each dose group and positive controls (P<0.05). Also, the difference between positive and negative controls was statistically significant (P<0.05). These results indicated that the Mongolian compound ingredients in Terunshudule did not cause chromosome fragmentation and spindle lesions in mice.5.Testicle chromosome aberration test in mice:we set up high- and low-dose groups, positive and negative controls. Twelve days after drug administration, testicles were separated, sliced and observed under a microscope. The results showed that the difference between each dose group and negative controls was not significant (P>0.05). However, the difference between each dose group and positive controls was significant (P<0.01). Also, the difference between positive and negative controls was significant (P<0.01). These results indicated that the compound in Terunshudule did not lead to chromosome abnormality and other genetic deformities.6.In vitro hemolysis test:we prepared red blood cell suspension using rabbit blood. Normal saline was used as negative control, and distilled water was used as the positive control, with different doses of compound Mongolian medicine added into the rest of the tubes. No macroscopic hemolysis was observed, which suggested that Terunshudule could be a reference for clinical injected medications.7.Allergy test:we used drug on rabbit skin after hair removed. The compound did not cause erythema, swelling and other allergic effects, indicating that this compound was non-allergic.