To explore the mechanism by which Tusizi (Cuscuta chinensis) flavonoids (CCF) improves the reproductive toxicity of Leigongtong (Tripterygium wilfordii) glycosides (GTW) in the male rats using Tandem Mass Tags (TMT) quantitative proteomics and bioinformatics methods.
Twenty-one male SD rats were randomly divided into a control group, a GTW model group and a CCF treatment group, and were orally administered with the corresponding drugs for 12 consecutive weeks. After the experiment, hematoxylin eosin staining was performed to evaluate the improvement effect of CCF on GTW-induced reproductive toxicity. TMT quantitative proteomics technology was used to evaluate proteins extracted from testicular tissue, analyzing differentially expressed proteins through bioinformatics methods and verifying key differentially expressed proteins screened by proteomics through western blot.
CCF can significantly improving the pathological damage of the testicular tissues of the rats. After intervention, a total of 29 differentially expressed proteins were detected, including 6 up-regulated proteins and 23 down-regulated proteins. KEGG pathway analysis showed that African trypanosomiasis, complement coagulation cascade system, biosynthesis of amphotericin, histidine metabolism and vitamin digestion and absorption were the main pathways by which CCF improved the reproductive toxicity of GTW. GO analysis showed that the differentially expressed proteins in CCF improving GTW reproductive toxicity were mainly related to collagen, plasma lipoprotein, protein oxidation, intrinsic coagulation pathways and heparin binding. COG annotation analysis showed that defense mechanisms, extracellular structures and the mechanisms by which CCF improved GTW reproductive toxicity were closely related. Dodder seed flavonoids can up-regulate the expressions of Apoa1and Tktl2, and down-regulate the expressions of Hba1, Aldh3a1, Pzp and Serpind1.
CCF can alleviate the testicular toxicity of GTW by regulating collagen synthesis, lipid metabolism, cell apoptosis and oxidative stress mediated by African trypanosomiasis, complement and coagulation cascade system, biosynthesis of ansamycins, histidine metabolism, vitamin digestion and absorption pathway.
| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |