Comparative Study Of The Influence On The Expression Of Spermatogenic Regulatory Gene SCF/c-kit,CREM,C-myc In Young And Adult Male Rats Intervened By Semen Cuscutae Flavonoids

Objective: By observing the impact of testicular spermatogenic cell cycle and the expression of critical regulatory gene SCF/c-kit,CREM and C-myc between adult and young rats which has been given drug withdrawal by GTW and Semen Cuscutae Flavonoids,to detect the intervention mechanism of flavonoids against reproductive damage caused by GTW on adult and young rats and to look for any difference in the expression of critical regulatory gene SCF/c-kit,CREM and C-myc between adult and young rats.Methods:Male SD in young rats 18(3 weeks old)and adult rats 18(9 weeks)are divided into six groups randomly: pups blank group,pups glycosides group,pups flavonoids group,adults blank group,adults glycosides group,and adults flavonoids group,each group have six rats.Blank groups are given 0.5% carboxymethyl cellulose 0.5ml/100 g daily,glycosides groups are given GTW 12.5mg/kg daily,while flavonoids groups are given GTW 12.5mg/kg and dodder flavonoids 0.2g/kg daily.The above dose are calculated according to the dosage of crude drugs.After gavaged for 8 weeks,rats are killed and its’ testicular tissue are taken out at the last gavage time after 1 hour.The testicular tissues were divided it into two parts aseptically,then mark and store them in vials,preserve them in-80 ℃ refrigerator.one part were used to measure the expression of SCF/c-kit,CREM and C-myc mRNA by Real-time PCR method.the other part were used to measure the protein expression of SCF/c-kit,CREM,C-myc by Western Blot method.Results: 1.The expression of rats’ spermatogenic regulating gene SCF in experimental groups under the infection of two drugs(1)In the mRNA expression of SCF,compared with each blank group,two glycosides groups are decreased,and there are significant differences in them(P<0.05).While two flavonoids groups are higher than the polyglycosides groups,and comparison among groups are statistically significant(P<0.05).The contrast among pups and adults glycosides groups and two flavonoids groups is not statistically significant(P>0.05).(2)In the protein expression of SCF,two glycosides groups are lower than themselves blank group,and the comparison are significant(P<0.05).Pups and adults flavonoids group are higher than themselves glycosides group,the compare between the pups groups is statistically significant(P<0.05).The contrast between pups groups and adults are not statistically significant(P>0.05).2.The expression of rats’ spermatogenic regulating gene c-kit in experimental groups under the infection of two drugs(1)In the mRNA expression of c-kit,two glycosides groups are lower than themselves blank group,and the comparison is significant P<0.05).Pups and adults flavonoids group are higher than themselves glycosides group,the compare between the adults groups is statistically significant(P<0.05).The contrast between pups groups and adults are not statistically significant(P>0.05).(2)In the protein expression of c-kit,two glycosides groups is lower than themselves blank group,and the comparison is significant(P<0.05).Pups and adults flavonoids group are higher than themselves glycosides group,the compare among groups havn’t significant(P>0.05).The contrast between pups groups and adults are not statistically significant P>0.05).3.The expression of rats’ spermatogenic regulating gene CREM in experimental groups under the infection of two drugs(1)In the mRNA expression of CREM,two glycosides groups are lower than themselves blank group,and the comparison are significant(P<0.05).Pups and adults flavonoids group are higher than themselves glycosides group,the compare between the pups groups is statistically significant(P<0.05).The contrast between pups groups and adults are not statistically significant(P>0.05).(2)In the protein expression of CREM,two glycosides groups are lower than themselves blank group obviously,pups and adults flavonoids group are higher than themselves glycosides group,the compare among groups are significant(P<0.05).The contrast between pups groups and adults are not statistically significant(P>0.05).4.The expression of rats’ spermatogenic regulating gene C-myc in experimental groups under the infection of two drugs(1)In the mRNA expression of C-myc,two glycosides groups are higher than themselves blank group,two flavonoids group are lower than themselves glycosides group,the compare among groups are statistically significant(P<0.05).The contrast between pups groups and adults are not statistically significant(P>0.05).(2)In the protein expression of C-myc,two glycosides groups is higher than themselves blank group,and the comparison is significant P<0.05).Pups and adults flavonoids group are higher than themselves glycosides group,the compare between groups havn’t statistically significant(P>0.05).The contrast between pups groups and adults are not statistically significant(P>0.05).Conclusion: 1.Semen Cuscutae Flavonoids can antagonize the inhibition of spermatogenic cell cadused by GTW,which may be due to repair the expression of spermatogenic regulatory gene SCF/c-kit and CREM,and the promotion of spermatogenic cell cadused by GTW through repair the expression of spermatogenic regulatory gene C-myc.2.GTW havn’t obviouly difference influence on the the expression of spermatogenic regulating gene SCF/c-kit、CREM、C-myc of young and adult male rats.3.Semen Cuscutae Flavonoids havn’t obviouly difference influence on the the expression of spermatogenic regulating gene SCF/c-kit、CREM、C-myc of young and adult male rats.