Regulation Of Inhibin Gene Vaccine Without Antibiotic Resistance Markers On Reproduction In Male Rats

Objective:To study the effects of inhibin gene vaccine enveloped by Salmonella in male rats’sperm maturation and development, biochemical indicators, testicular apoptosis and apoptotic protein expression and reproductive related hormones to investigate the role and mechanism of new gene immunization against inhibin on male rat reproductive regulation. Methods:Using different concentrations of inhibin gene vaccine to actively immunize male rats, testing the changes of inhibin antibody level during the entire experimental periods, the coefficient of rats reproductive organs, the activity of testicular GGT, AKP, LDH, α-glucosidase and the content of carnitine; the carnitine levels and a-glucosidase activity of epididymis; the activity of prostatic acid phosphatase; the fructose content of seminal vesicle, the percentage of epididymal sperm deformity, expression and apoptosis of testicular apoptotic proteins and FSH, LH and Testosterone levels in the experimental period.Results:(1) Inhibin antibody P/N values in the immunized group were significantly higher than the experimental group, the second week after the primary immunity the P/N values of medium-group was significantly higher than in the rest four groups(P<0.01), high-dose group was significantly higher than in low-dose group, empty plasmid group and the control group(P<0.01); in the first week after the secondary immunity the P/N values of high-dose group and medium-group was significantly higher than in the rest three groups(P<0.01), medium-group was significantly higher than in the high-dose group(P<0.05). In the second week after the immunity the positive rate of the medium-group was significantly higher than the low-dose group(P<0.01), in the third week after the immunity the positive rate of the medium-group was significantly higher than the low-dose group(P<0.01). In the first week after the secondary immunity the positive rate of the medium-dose and high-dose group was85.71%and57.14%respectively.(2) The coefficient of testis in the medium-dose group was significantly higher than that the empty plasmid group (P <0.05), between each group the coefficient of epididymis, prostate and seminal vesicles was not significant.(3) Testicular GGT activities in the low-dose and medium-dose group were significantly higher than those in the control group (P<0.01); testicular AKP activity of low-dose group was significantly higher than that in the empty plasmid group (P<0.05), testicular ACP activity in medium-dose group was significantly lower than that in the control group (P<0.05); testicular LDH activity of the high-dose group was significantly higher than that in in the low-dose group (P<0.05); testicular a-glucosidase activity of the medium-dose group was significantly lower than that in the control group (P<0.01). Epididymal a-glucosidase activity of low-dose group was significantly higher than that in the control group (P <0.01). The activity of prostatic acid phosphatase in the medium-dose group was significantly lower than that in the control group (P<0.01).(4) The rate of sperm deformity in the high-dose group was significantly lower than that in the control group (P<0.01), and the number of sperm in the high-dose group was significantly lower than that in the control group (P<0.05); testicular apoptosis in the medium-dose group was significantly higher than that in the control group (P<0.01); the expression of Bax protein and Bcl-2protein have no significance differnces in all groups(P>0.05).(5) The FSH level of mediun-dose group was increased than the control group after the immunization:the second week after the primary immunity the FSH level in medium-dose group was significantly increased than in the empty plasmid group(P <0.01), medium-dose group was significantly increased than in the control group(P <0.05). LH level in the control group was significantly increased than that in the other group:the first week after the secondary immunity the LH level of medium-dose group and low-dose group was significantly lower than the control group(P<0.01), high-dose group and empty plamid group was significantly lower than the control group(P<0.05). Testosterone levels in each dose group significantly increased than the control group:the second week after the secondary immunity the T level of the high-dose group and the low-dose group was significantly higher than the control group(P<0.05); the first week after the third immunity the T level of the midum-dose group and the low-dose group was significantly higher than the control group(P <0.05).Conclusion:Inhibin gene immunization can increase inhibin antibody levels in the male rats, promote the development of rat testis and lower the percentage of rat sperm deformity, also can promote testicular apoptosis and increase conents of FSH, testosterone but reduceLH contents in the male rats.