Preliminary Evaluation On Toxic Reaction Of Tripterygium Polyglycoside In Bi Syndrome Rats

ObjectiveUsing the rats with adjuvant arthritis as Bi syndrome animal model, to evaluate the toxic reaction of rats with Bi syndrome induced by Tripterygium Wilfordii Polyglycoside (TWP) preliminarily from the blood biochemistry, pathology , immunology and metabolomics.MethodsMale SD rats were selected as experimental object, and 40 rats were induced as AA model rats with the method of subcutaneous injecting freund ' s complete adjuvant in right hind. 40 AA model rats and 40 normal rats were randomly divided into 4 groups seperately, ten in each group, including model blank control group, groups with low, middle and high dosages, normal blank control group and groups with low, middle and high dosages. Blank group rats were given physiological saline; low, middle and high dose group rats were administered with TwP of 7mg/kg (equivalent to the clinical therapeutic dose of people), 70mg/kg, 105mg/kg seperately, once a day, observing and measuring the weight and articular swelling degree of the rats. Animals were killed after 2 weeks, then taking blood for biochemical and blood routine tests; measuring the weight of spleen, thymus, liver, kidney, testis, epididymis and prostate, and calculating the organ coefficient; observing the pathological damage of the ankle joint and organ with light microscope; detecting the changes of T lymphocyte subsets by streaming technique, observing the expression of endothelial nitric oxide synthase (eNOS), Neuronal nitric oxide synthase (nNOS) and inducible nitric oxide synthase (iNOS) in testicular tissue by Immunohistochemistry, testing the change of iNOS, TAOC in plasma by colorimetric method, inspecting the situation of small molecular in rat plasma by metabolomics method.Results 1. The degree of ankle swellingThe ankle swelling degree in inflammation side and Secondary side of the rats in the model group was increased significantly than that of the normal group. After giving medicine, ankle swelling degree of rats in the model low-dose group showed a decreasing tendency compared to model group.2. Organ coefficientLiver coefficient was increased with dosage increasing in the model group, no significant changes in the normal group. Testis coefficient were increased with decreasing dose in both normal group and model group. Spleen, kidney, epididymis and prostate coefficient can not be compared because they did not conform to generalized linear models.3. Blood biochemical and routine tests resultsAspartate aminotransferase (AST) showed an increasing tendency with dosage increasing in the normal group, while no change in the model group. Alkaline Phosphatase (ALP) showed a decreasing tendency with dosage increasing in the model group, while i had an increasing tendency n the normal group. There was no significant effect on Creatinine (CRE) in different groups with different doses. However, there was difference in UAC. The normal group was significantly increased compared to the model group, especially at middle dose. Glucose (GLU) in the model group had no change with dosage increasing, while had an increasing tendency in the normal group.There was no significant effect on Hemoglobin (HGB), Red blood cell disposition width (RDW), Lymphocyte (LY) and Mean corpuscular volume (MCV) in different groups with different doses. PLT had an increasing tendency with dosage increasing in the model group, while there was no change in the normal group.4.T cell subsets in peripheral bloodCompared with the model group, the normal group showed differences in peripheral blood CD3. With the dosage increasing, the value of CD3 had decreasing trend in the normal group, nevertheless it unchanged in the model group. 5.Expression of iNOS,eNOS and nNOS in testicular tissueAt low dose, there was no significant difference between the normal group and the model group. At middle dose, the expression of eNOS and nNOS in the model group was increased compared to the normal group, while vice versa in expression of iNOS. At high dose, both the expression of eNOS and nNOS decreased significantly, while iNOS increased obviously.6. iNOS and TAOC in plasma testing result.INOS cloud not be compared because it did not conform to generalized linear models. Total anti-oxide capability (TAOC) have the elevation tendency with dosage increasing in the normal group, while there was no change in the model group.7.Metabolomics Study ResultThere was no significant effect on Lysophosphatidyl choline (LPC) C16:0 and LPC C18:1 in different groups with different doses. With the dose change, Ursodeoxycholic acid (UDCA) and Chenodexycholic acid (CDCA) had the decreasing trend.8. Light microscope resultsJoint : Pathological change of the joints in the model low dose group was slighter than model blank control group.Organ: There was no abnormal change of liver in rats of normal group, while model rats showed pathological change. The rats in Normal middle, high-dose group and model middle, high-dose group all had renal pathological change, and model rats were more serious at the same dosage level. Pathological change of testicular in model middle group was slighter than in normal middle group. At high dose, both group showed serious pathological changes.Conclusion1. TWP of low dose could play the role of treatment through relieving the degree of ankle swelling in AA rats. The rats showed no obvious toxic reaction in both the normal and the model groups.2. Some indicators showed abnormity in the normal and model middle dose groups, whereas the normal middle dose rats were observed stronger toxic reaction than the model.3. The rats administered high dose TWP showed severe toxic reaction in normal and model groups. It indicated that TWP administered an excess dosage could induce severe toxic reaction in rats no matter physiological or pathological conditions.