Objective To study the protective effects on ovarian function by caloric restriction (CR) and its mechanism.Methods Thirty female C57BL/6 mice of 8 weeks old were randomly divided into two groups,including ad libitum (AL) group and caloric restriction (CR) group.The general situation and ovarian function of those mice were compared and evaluated.Ovarian follicles were counted by hematoxylin-eosin staining.Anti-Miillerian Hormone(AMH) mRNA expression of the ovary were detected by using real-time PCR.The concentrations of serum estradiol,progesterone of the mice were measured by ELISA.And the fertility of mice by mating trials were evaluated,SIRT3,Hypoxia inducible factor 1α (HIF-1α) and catalase (CAT) mRNA expression of the mice ovaries were detected by Real-Time PCR.Results The total follicles were 546 in CR mice and 286 in AL mice.The proportion of primordial follicles were 38.6％ (211/546)in ovaries of CR mice and 29.4％ (84/286)in ovaries of AL mice,which reached statistical difference.The proportion of atretic follicles 5.3％ (29/546) in ovaries of CR mice,compared with 16.8％ (48/286) in AL mice,was significantly decreased (P ＜ 0.05).The AMH mRNA expression in CR mice ovaries was 3.37 times of that of AL mice (P ＜ 0.05).The serum concentration of estradiol in CR mice was up to (5.3 ± 1.6) pmol/L,which was much higher than (3.6 ± 1.6) pmol/L in AL mice.While,the progesterone concentration of (0.4 ±0.3) nmol/L in CR mice was lower than (1.4 ± 0.8) nmol/L in AL mice (P ＜ 0.05).Fertility and survival of offsprings were both improved in CR mice.The expression level of SIRT3 mRNA in CR mice ovary was 1.39 times,CAT was 1.55 times and HIF-1 α was 0.31 times of those in AL mice (P ＜ 0.05).Conclusions Caloric restriction can delay the ovary aging process through reduce follicle depletion by suppressing follicle recruitment and ovulation.The function of ovarian reserve and reproductive endocrine was effectively protected.Caloric restriction can reduce the incidence of follicular atresia,its mechanism might be associated with anti-oxidative stress.

ObjectiveTo explore the mechanism of Si Junzitang in regulating the expression of NKG2A to affect the anti-colon cancer function of natural killer （NK） cells. MethodNK cells isolated from healthy honors were cultured and used to construct the three incubation models of NK cells， human colon cancer HCT116 cells， and NK cells + HCT116 cells （co-incubation）. real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was conducted to determine the mRNA levels of natural killer group 2 member A （NKG2A） and interleukin （IL）-15 in NK cells， as well as the mRNA level of histocompatibility leucocyte antigen E （HLA-E） in HCT116 cells. The secretion of IL-15 was detected by enzyme-linked immunosorbent assay （ELISA）. Methyl thiazolyl tetrazolium （MTT） assay was employed to determine the applicable concentration of IL-15 and test the effects of Si Junzitang and IL-15 on the activities of NK cells and the HCT116 cells in the co-incubation model. The effects of Si Junzitang and IL-15 on the mRNA levels of NKG2A in NK cells and HLA-E in HCT116 cells were detected by Real-time PCR. Monalizumab （M， anti-NKG2A mab） was used to block the NKG2A-HLA-E pathway in co-incubation model， and then the proliferation of HCT116 cells was detected by MTT assay. ResultThe interaction of NK cells and HCT116 cells up-regulated the mRNA levels of NKG2A in NK cells and HLA-E in HCT116 cells （P<0.05）， as well as the expression level and secretion of IL-15 （P<0.05）. Compared with the blank group， Si Junzitang and Si Junzitang + IL-15 promoted the proliferation and improved the anti-colon cancer function of NK cells （P<0.01）. Furthermore， they down-regulated the mRNA levels of NKG2A in NK cells and HLA-E in the HCT116 cells co-incubated with NK cells （P<0.01）. M and IL-15 + M inhibited the proliferation of HCT116 cells compared with the groups without M （P<0.01）. ConclusionThe interaction of NK cells and HCT116 cells can induce activation of NKG2A-HLA-E pathway to impair NK cell function. Si Junzitang can inhibit the activation of NKG2A-HLA-E pathway to restore the anti-colon cancer function of NK cells.