Effect Of Acupuncture And Moxibustion On The Contents Of γ-H2AX And Ku70 In Testis Of CTX Mice

Objective In this study,a mouse testis injury model was established by cy clophosphamide(CTX)intraperitoneal injection.By observing the changes of te sticular histopathology and the contents of phosphorylated histone γ-H2 AX and nuclear Ku70 protein in testicular tissues before and after acupuncture intervent ion,the mechanism of acupuncture and moxibustion on the repair of testicular injury induced by CTX was preliminarily revealed,from the perspective DNA t esticular cell injury can cause spermatogenesis disorder.Methods No specific pathogen(SPF)grade male Kunming species(KM)mouse 48,age 7 weeks.Weighing after 3 days of adaptive feeding,randomly di vided into blank group(A),model group(B),acupuncture group(C)and moxi bustion group(D)according to weight stratification.Each group was twelve.T he mouse in B,C and D group were injected with CTX solution(concentration 5mg/m L)intraperitoneally at the same time daily at m L/g/dose of 0.02 for thr ee consecutive days,to prepare a model of mouse testicular injury.As a contro l group,the blank group of mice were accompanied by intraperitoneal injection of the same dose of 0.9% Nacl solution 0.02 m L/g for three consecutive days.After modeling 24 h,one mouse was randomly selected from each group to be executed,and the testicular tissue was removed by laparotomy for pathological observation to verify the success of the mouse testicular injury model.When t he model was prepared,acupuncture and moxibustion were given to C and D g roup of mice in order,and the acupoints were treated with "Dazhui" and bilate ral "Geshu","Shenshu" and "Zusanli";A and B group of mice were only acco mpanied by fixation and not treated.Once a day for five consecutive days.Tw o mice were randomly selected from each group the next day after treatment,a nd the testis tissue was fixed with 4% neutral paraformaldehyde to be made in to pathological sections.The remaining mice were collected and stored in-80℃ refrigerator.The γ-H2 AX and Ku70 contents were detected by enzyme-link ed immunoassay(ELISA).SPSS 23.0 for Windows was used to analyze the ex perimental data.Results1 Effects of acupuncture and moxibustion on testicular tissue morphology of CTX model mice: before treatment,the morphology and structure of testicul ar tissue in the blank group were basically complete,the spermatogenic cells w ere stratified,arranged neatly,and the spermatogenesis was good;the testicular tissue of the mice in the CTX model group was incomplete in morphology,th e spermatogenic epithelium was damaged seriously,the number of spermatogeni c cells was decreased,the arrangement was disordered,the cell space was enlar ged,the spermatogenesis was poor,and some of them showed that the nucleus was thick,vacuolar degeneration,nuclear fixation and so on.After 5d of treat ment,compared with the model group,the morphology of testicular tissue of m ice in acupuncture group and moxibustion group was relatively complete,the d amage of spermatogenic epithelium was alleviated,the number of spermatogenic cells was relatively increased,the number of cell vacuolar degeneration and n uclear contraction was reduced,and the sperm production was better,and the s ituation of moxibustion group was obviously better than that of acupuncture gr oup.2 The effect of acupuncture on γ-H2 AX and Ku70 content in testicular tis sue of CTX model mice: Compared with the blank group,the γ-H2 AX and Ku70 content in the CTX model mice groups increased significantly(P<0.05);Com pared with the model group,the γ-H2 AX and Ku70 contents of acupuncture gr oup and moxibustion group decreased significantly(P<0.05);The γ-H2 AX and Ku70 contents in moxibustion group were lower than those in acupuncture grou p(P<0.05).Conclusion1 Acupuncture and moxibustion can improve the testicular injury caused by cyclophosphamide,and promote the recovery of normal morphology and struct ure of testicular tissue by repairing the spermatogenic epithelium of seminiferou s tubules,promoting the increase of endogenous spermatocyte levels in the lum en,and reducing the pathological manifestations such as nuclear contraction and vacuolar degeneration.2 Acupuncture and moxibustion improve testis cell DNA double-strand brea ks by down-regulating γ-H2 AX and Ku70 expression in testis tissues of CTX model mice,thereby repairing CTX damaged testis cells and alleviating CTX m ouse testis injury.3 The effect of moxibustion on down-regulation of γ-H2 AX and Ku70 exp ression in testicular tissue is more significant than that of acupuncture,and the n it is more beneficial to repair CTX induced injury of testicular cell DNA.