Objective To observe the effect of warm needle moxibustion(WNM)

mild moxibustion and electroacupuncture(EA)of"Shenshu"(BL23)and"Guanyuan"(CV4)on serum gonadal hormone levels and related gene expression in the hypothalamus and testis in aged rats

so as to explore their underlying mechanisms in anti-aging.Methods Forty male SD male rats were randomly divided into aged control

WNM

mild moxibustion

EA and medication groups(n = 8 in each group)

and other 8 male young adult SD rats were used as the youth control group.WNM(acupuncture plus moxibustion)

mild moxibustion or EA(2 Hz/100 Hz

1 mA)was separately applied to CV4 and bilateral BL23 for 15 min

once daily

5 days a week for 8 weeks.Rats of the medication group received subcutaneous injection of testosterone propionate injection(7 mg/kg)

once every 3 days

and those of the aged and youth groups received the same dose of 0.9% NaCl solution for 8 weeks.The levels of serum testosterone(T)and gonadotropin-releasing hormone(GnRH)were measured by ELISA.The expression of Kisspeptin(KISS-1)

G protein-coupled receptors 54(GPR54)

RFamide-related peptide-3(RFRP-3)and GPR147 mRNAs in the hypothalamus and GnRH-R

GPR54 and GPR147 mRNAs in the testis tissues were detected by quantitative real-time PCR.Results The levels of serum T and GnRH

and the expressions of KISS-1

GPR54

RFRP-3 and GPR147 mRNAs in the hypothalamus and GnRH-R and GPR54 mRNAs in the testis in the aged rats were significantly lower than those in the youth rats(P<0.05)

while the expression level of GPR147 mRNA of the testis tissue was obviously higher in the aged group than that in the youth group(P<0.05).Following the treatment

the lowered serum T and GnRH contents

the down-regulated expression levels of hypothalamic KISS-1

GPR54

RFRP-3 and GPR147 mRNAs and testicle GnRH-R and GPR54 mRNAs

as well as the up-regulated testicle GPR147 mRNA expression in the aged rats were reversed in the WNM

mild moxibustion and EA groups(not the medication group except serum T)in comparison with the aged control group(P<0.05).The therapeutic effects of WNM

mild moxibustion and EA were significantly superior to those of medication in up-regulating serum GnRH content and expression levels of KISS-1

GPR54

RFRP-3 and GPR147 mRNAs in the hypothalamus and GnRH-R

GPR54 and GPR147 mRNAs in the testis(P<0.05).No significant differences were found among the WNM

mild moxibustion and EA groups in serum T and GnRH contents

and the expression levels of KISS-1

GPR54

RFRP-3 and GPR147 mRNAs in the hypothalamus and GnRH-R and GPR54 and GPR147 mRNAs in the testis(P>0.05).Conclusion The WNM

mild moxibustion and EA therapies have an anti-aging effect by increasing serum T and GnRH levels in aged rats

which is possibly related to their effects in up-regulating the expression of hypothalamic KISS-1

GPR54

RFRP-3 and GPR147 mRNAs and testicle GnRH-R and GPR54 mRNAs

as well as down-regulating testicle GPR147 mRNA expression.The therapeutic effect of WNM

mild moxibustion and EA is evidently superior to that of medication.