Effects Of RNA Interference UL27 Gene On Replication Of HSV-2 In Mice

Objective: The effects of UL27 gene knockdown on replication of HSV-2 will be assessed by implanting UL27 sh RNA recombinant expression vector into the mouse model of genital herpes(GH)in this study.Methods: GH mice model was manufactured by means of vaginal embolism injection.Thirty GH model mice were divided into three groups(10 mices in each group): UL27 sh RNA treatment group,negative sh RNA group and liposome suspension as black control group.The sh RNA plasmids were respectively transfected into mice vaginal epithelium by lipofectamine 2000 at three times everyday(4 hours interval)for seven days.Recording the mice vulvar symptoms at 3,5,7 day after treatment.Cotton dips in vaginal secretions before dissecting the mice.Dissecting the mice after prescription to observe curative effect.HE staining techniques were used to observe the changes of vaginal tissue in pathology morphology.The expressions of UL27 m RNA and glycoprotein protein were detected by RT-PCR and Western blotting methods.Results:(1)By vaginal embolism injections transfect 0.1ml TCID5010-5 virus to mouse vaginal epithelium,vulva appeared swollen and secretion increased.There were blisters in skin and edema in the cell.(2)Three days after the treatment,the vulvar lesion score in UL27 sh RNA treatment group was obviously lower than negative sh RNA and blank control groups(P<0.05),while there was no significant difference in the score between negative sh RNA group and the blank control group.(3)Compared with other groups,the virus drops significantly decreased in UL27 sh RNA treatment group(P<0.05).Meanwhile,there was no significant difference in the virus drops between negative sh RNA group and the blank control group.(4)The results of pathological observation showed that the UL27sh RNA treatment effectively inhibited the infiltration of inflammatory cells in mice than other groups.(5)RT-PCR results demonstrated that compared with blank control,UL27 sh RNA treatment significantly inhibited its m RNA expression(P<0.05),while negative sh RNA treatment had no effect on UL27 m RNA expression.(6)UL27sh RNA treatment inhibited glycoprotein protein expression in vaginal tissue compared with the other two groups(P<0.05),while there was no effect on protein expression between negative sh RNA group and the blank control group.Conclusion: The UL27 sh RNA recombinant expression vector could inhibit UL27 gene and protein expression in female BALB/c mice with HSV-2 infection.Moreover,the knockdown of UL27 gene obviously alleviated infiltration of inflammatory cells in vaginal epithelium.