| Muskrat(Ondatra zibethicus Linnaeus)also called aone mink.It is the only species of muskrat subfamily of voles.Male muskrat have a pair of musk glands.Musk glands can secrete aromatic white musk on breeding season.The main medicinal components of muskrat musk is muscone,and it is similar to natural musk.So it has great potential to be a substitute for natural musk.Muskrat compared to other murine,prostate tissue is very developed,and the experiments have proved that intraperitoneal injection of muskrat musk can result in prostate tissue increased.So it not only has the effect of pheromones,may also have to adjust their development role.Through RNA-seq sequencing technology in this experiment,the transcriptional library of muskrat secreting stage and non-secreting stage of prostate tissue was sequenced.Through the bioinformatics from the overall level of muskrat prostate development are discussed.At the same time,this experiment established into primary muskrat prostate fibroblast like cells culture method,successfully constructed a cell model of muskrat prostate fibroblast like cells,and has been proved the feasibility of the cell culture in vitro.Combined with the results of RNA-seq sequencing and the cell model of primary culture,conducted the muskrat intervention trial of primary cells.The third generations of muskrat cultured prostate fibroblast like cells were divided into 2 groups,10 ?g/ml of muskrat musk group denoted as A1 group,0.1%DMSO control group was recorded as group B1.Experimental results show that the transcriptional level of Bcl-2 in group A1 was about 4 times higher than that in group B1.The transcriptional level of Bax gene was lower than that in B1.Muskrat musk can increase the transcription level of Bcl2 gene,reduce the transcription level of Bax gene,enhancing the ratio of Bcl2/Bax to anti-apoptosis.For a preliminary study on the mechanism for adjusting the Bcl2/Bax ratio of muskrat musk,this experiment using HEK293T cells,and the intervention using muscone.The HEK293T cells were divided into 5 groups,the group of 3?g/ml muscone was set as group A2;the group of 5 ?g/ml muscone was set as group B2;the group of 7 ?g/ml muscone was set as group C2;the group of 9 ?g/ml muscone was set as group D2;the group of 0.1%DMSO was recorded as group E2.Set three biological replicates in each group.Compared with group E2,the transcription level of NF-?B gene in A2,B2,C2 and D2 groups were 1.39 times,1.53 times,1.72 times and 1.78 times of E2,respectively;the transcriptional level of the PI3K gene was 1.41 times,1.58 times,1.71 times,and 1.79 times of the E2 group;the transcriptional level of the ERK gene was 1.27 times,1.39 times,1.53 times and 1.77 times of the E2 group;the transcriptional level of the MEK gene was 1.2 times,1.29 times,1.32 times and 1.38 times of group E2;the transcriptional level of the p53 gene was 1.08 times,1.11 times,0.99 times and 1.1 times of group E2.The results showed that Muscone increased the transcriptional level of NF-?B,PI3K,MEK and ERK genes,and they have a significant dose-effect relationship with the concentration of Muscone.But there was no obvious effect on the p53 gene.Therefore,it is presumed that the change of the Bcl-2 and Bax gene ratio of the Muscone regulated cells may be accomplished by regulating these signal pathway.Predicted by RNA-seq sequencing and drug intervention cell experimental results,muskrat musk itself not only plays the role of pheromones,also may regulate the growth and development of their own,for example,can promote the development of the prostate.The muscone and other lipophilic odorant molecules with the blood circulation to the prostate tissue.In this process,the protein encoded by the c75134-g2 transcript acts as a lipophilic molecular carrier to help it bind to the receptor.By regulating the transcriptional level of the Bcl-2 gene,it plays a role in anti-apoptosis and regulates the development of the prostate tissue. |
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