Effect Of Xiongcan Yishen Recipe On The Expression Of Circadian Clock Genes In Testis Of LOH Rat

Objective: To investigate the effect of Xiongcanyishen formula on the expression of circadian clock gene in testicular tissue of LOH rats.Methods: Forty-eight 8-week-old male SD rats were randomly divided into normal group,model group,testosterone propionate group,Xiongcanyishen formula low-dose,medium-dose and high-dose groups.Model group,testosterone propionate group,male Chanyishen formula low-dose,medium-dose and high-dose groups were intraperitoneally injected with 480mg/(kg·d)D-gal,and continuously injected for 56 d,and normal group were intraperitoneally injected with the same volume of normal saline every day.After successful modeling,Xiongcanyishen low-dose,medium-dose and high-dose groups were given 10.4,20.8 and 41.6g/(kg·d)body weight of Xiongcanyishen decoction by gavage,respectively.Rats in testosterone propionate group were intramuscularly injected with 5.21mg/(kg·d)body weight of testosterone propionate,3 times a week;Normal group and model group were given the same volume of distilled water intragastric administration for 28 days.After the experiment,blood samples were centrifuged and supernatant was taken to detect the level of testosterone(T)by ELISA;testicular tissues were taken for Western blot and RT-q PCR to detect the protein expression levels and m RNA expression levels of BMAL1,NR1D1,PER2,CRY1,St AR and CYP11A1,respectively.Results: The serum testosterone and m RNA and protein expression levels of BMAL1,NR1D1,PER2,CRY1,St AR and CYP11A1 in testis in model group were significantly lower than those in normal group(P <0.05).Compared with model group,serum T in testosterone propiate group and Xiongcanyishen formula low-dose,medium-dose and high-dose groups was significantly increased(P<0.05),while St AR m RNA expression level,Cyp11a1 m RNA expression level and protein expression level in testicular tissue of Xiongcanyishen formula high-dose group were significantly increased(P<0.05).The expression level of St AR protein in testicular tissue of Xiongcanyishen formula in low,medium and high dose groups was significantly increased(P<0.05),and the protein expression levels of BMAL1 and NR1D1,PER2 and m RNA in testicular tissue of testosterone propiate group and Xiongcanyishen in low,medium and high dose groups were significantly increased(P<0.05).The expression level of BMAL1 m RNA and CRY1 protein in testicle tissue of Xiongcanyishen formula medium and high dose groups was significantly increased(P<0.05),and the expression level of NR1D1 m RNA in testicle tissue of Xiongcanyishen formula low,medium and high dose groups was significantly increased(P<0.05).The expression level of CRY1 m RNA in testicular tissue of testosterone propionate group,Xiongcanyishen formula medium and high dose groups was significantly increased(P<0.05).Conclusion: Xiongcanyishen formula up-regulates the mRNA expression levels and protein expression levels of BMAL1,NR1D1,PER2,Cry1 in testicular tissue of LOH rats,regulates the m RNA and protein expressions of cholesterol transporter St AR and testosterone synthetase CYP11A1,and improves serum testosterone level,which may be one of the mechanisms of Xiongcanyishen formul’s intervention in LOH.