Targeted microbubble disruption technology can reversibly destroy the blood-testicular barrier
Objective To explore the effect of ultrasound-targeted microbubble disruption technol-ogy(UTMD)on the blood-testis barrier of rat testis.Methods Forty-eigg adult male SD rats were ran-domly(randomization generator method)divided into control,ultrasound(US),ultrasound+low concen-tration of microbubbles(US+LM),and ultrasound+high concentration of microbubbles(US+HM)groups under different experimental conditions.Immunoprotein blotting for tight junction protein(Occul-din,ZO1),actin regulatory protein(Forminl,Arp3)microtubule protein regulatory protein(EB1),and cytoskeletal protein(F-actin)expression in spermatocytes were detected by immunocytofluorescence,and the changes in the number and distribution of blood-testis barrier component proteins were observed after 6 h.The degree of BTB damage was semi-quantified by the ratio of biotin diffusion distance(D)to the ra-dius of the seminiferous tubules(R)x 100％.Comparisons between two groups were performed by t-test,and comparisons between multiple groups were performed by one-way ANOVA.Results D/R ratio,con-trol group:0％;US group:(37.97±0.32)％;US+LM group:(80.70±1.70)％;US+HM group:(98.33±1.44)％(US group vs.US+LM group,t=-42.639,P＜0.01;US group vs.US+HM group,t=-71.033,P＜0.01;US+LM group vs.US+HM group,t=-13.695,P＜0.01).F-actin:control group:long straight strip,oriented from basement membrane to the center of seminiferous tubules;US group:mildly curved strip,oriented slightly away from basement membrane to the center of seminiferous tubules;US+LM group:moderately curved strip,oriented significantly away from basement membrane to the center of seminiferous tubules;US+HM group:broken short sheet,not oriented.EB1/β-actin:control group:(0.95±0.01);US group:(0.72±0.02);US+LM group:(0.54±0.09);US+HM group:(0.44±0.01)(control group vs.US group,t=-20.602,P＜0.01;control group vs.US+LM group,t=-7.719,P＜0.01;control group vs US+HM group,t=-57.386,P＜0.01).Conclusion UTMD may mediate the reversible disruption of testicular barrier through EB1,F-actin.
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