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To search for specific protein makers and target proteins for intervention with Yimusake Tablets (YT) in the penile tissue of rats with ED induced by compound cold stress and explore the molecular mechanisms underlying the development and progression of ED.Eighty adult male rats were screened and divided into three groups, normal control (n = 10), ED model control (n = 15) and YT intervention (n = 15). The model of compound cold stress-induced ED was established in the latter two groups, and meanwhile the animals in the YT intervention group were treated with oral YT for 2 weeks. After that, proteins were extracted from the penile tissues of the rats for screening and identification by iTRAQ labeling combined with LC-MS-MS proteomics, and the IPA bioinformatics software was used for analysis of differentially expressed proteins.A total of 48 differentially expressed proteins were identified from the penile tissue of the ED model controls, of which 18 were associated with endothelial function, 5 with smooth muscle activity and 4 with inflammation, involving the biological processes of glucose metabolism and alcohol catabolism and the signaling pathways of glucose metabolism, calcium and RXR activation. In comparison, 29 differentially expressed proteins were identified from the rats in the YT intervention group, of which 5 were associated with endothelial function, 1 with smooth muscle activity and 4 with inflammation, involving the biological processes of glucose metabolism, vasodilation and acute-phase response and the signaling pathways glucose metabolism, RXR activation and acute-phase response. Seven ED-associated candidate biomarkers were obtained from the differentially expressed proteins in the ED model control and YT intervention group, including Collagen alpha-1(III) chain（COL3α1）, Collagen alpha-1(I) chain（COL1α1）, Collagen alpha-2(I) chain（COL1α2）, Glyceraldehyde-3-phosphate dehydrogenase（GAPDH）, T-kininogen 1（MAP1）,Biglycan（BGN）, and Myosin-11（MYH11）.Changes of vascular endothelial and smooth muscle functions in the penile tissue are likely to be the key mechanisms underlying the development and progression of compound stress-induced ED, which is also associated with inflammation as well as the interaction of the identified differentially expressed proteins and their participation in the relevant signaling pathways. The 7 proteins obtained can be used as the markers of compound stress-induced ED in the rat penile tissue, of which MAP1, GAPDH, BGN and MYH11 may serve as target proteins for YT intervention.

To determine the expressions of calcitonin gene-related peptide (CGRP) and vasoactive intestinal peptide (VIP) in the penile tissue of the ED rat model and explore their action mechanisms.An ED model was established in 44 mature male SD rats by feeding them on a spinach + coriander diet in a cold-wet environment and another 10 were taken as normal controls. Then the model rats were randomly divided into an ED model control group (n = 15) treated by gavage of distilled water in the same modeling environment, a spontaneous recovery group (n = 15) treated by gavage of distilled water in the normal environment, and a medication group (n = 14) treated intragastrically with Yimusake Tablets at 250 mg/kg qd. After 2－3 weeks of intervention, the expressions of CGRP and VIP in the penile tissue were detected by immunohistochemistry and Western blot.Immunohistochemistry showed that, after 2 weeks of intervention, both the expressions of CGRP and VIP in the rat penile tissue were significantly lower in the ED model control (150.0 ± 43.3 and 36.4 ± 13.1) and the spontaneous recovery group (165.9 ± 40.7 and 67.5 ± 29.0) than in the normal control (227.3 ± 42.5 and 175.0 ± 45.6) (P0.05), but remarkably higher in the medication group (255.0 ± 38.7 and 167.5 ± 42.6) than those in the ED model control and spontaneous recovery groups (P0.05).The expressions of CGRP and VIP were significantly down-regulated in the ED rat model, and Yimusake Tablets improved ED by up-regulating their expressions.