Proteomic Identification Of Differentially Expressed Nuclear Proteins In Vaccaria Segetalis-treated Dairy Cow Mammary Epithelial Cells

Studies of the regulation of the dairy cow lactation to solve the problems, as low milk yields and poor quality milk, which restrict the development of China dairy cow cultivation, have important theoretical and practical significance. Vaccariae segetalis is one of the most widely used Chinese traditional herbs to promote lactation; it has been widely applied in clinical and animal husbandry. It may have estrogen activity, which can activate lactation signal way, and regulate the synthesis of the milk protein, lactose and milk fat, and then increases lactation. But, the mechanism of Vaccariae segetalis promoting lactation is not clear; especially how to regulate genetic expression of lactogenesis has not been reported.In the experiments, CASY(?)-technology was used to study the proliferation and viability of the dairy cow mammary gland epithelial cells (DCMECs) at different time after Vaccariae segetalis treatments; protein expression profiles were analyzed by2-DE coupled with MALDI-Q-TOF MS/MS analysis; and such alterations were further confirmed by RT-PCR. The results showed that:①It was proved that Vaccariae segetalis could increase the proliferation and viability of primary cultured DCMECs significantly (P<0.05), and at24h was the best.②A2-DE separation system for proteome of nucleuar proteins of DCMECs was successfully established.③Five proteins, proteins ethylmalonic encephalopathy1(ETHE1), vesicle amine transport protein1homolog(VAT1), parkinson disease protein7homolog(Protein DJ-1), proteasome subunit alpha type-2(PSMA2) and SUMO-activating enzyme subunit1(SAE1), displayed significant expression changes upon treatment of the water decoction of Vaccariae segetalis, and such alterations were further confirmed by qRT-PCR.④Five genes were different expressed in these three dairy cows of lactation level. The highest expression level was in tissues of high-yielding dairy cows, followed by low-yielding cows, the lowest were perinatal cows. The results could be drawn from the above, these five genes associated with lactation.In conclusion, Vaccariae segetalis can increases proliferation of DCMECs significantly. It is the first time by proteomics methods to illustrate the mechanism of Vaccariae segetalis on the DCMECs, and reveal the proteomics bases of the DCMECs with Semen Vaccaria, further the specific mechanism of each protein needs to be studied.