ABSTRACT
ObjectiveTo observe the effect of Zuoguiwan on ovarian reserve in the female offspring rat model of prenatal stress （PS） and explore the mechanism based on Toll-like receptor 4/nuclear factor-κB p65 （TLR4/NF-κB p65） signaling pathway. MethodThirty-two pregnant rats were prepared and randomized into four groups （n=8）： control， model， Zuoguiwan （18.9 mg·kg-1）， and vitamin E （1.44 mg·kg-1）. Except the control group， the other three groups were subjected to chronic unpredictable mild stress （CUMS） from day 11 of pregnancy， and the modeling was accompanied by gavage with corresponding drugs until delivery. The PS model was evaluated by the sucrose preference test， open field test， and serum corticosterone （CORT） level. The estrous cycle was monitored and the morphological changes in the ovarian tissue were observed. The serum levels of estradiol （E2）， luteinizing hormone （LH）， follicle-stimulating hormone （FSH）， and anti-Mullerian hormone （AMH） in the 75-day-old offspring rats were measured by enzyme-linked immunosorbent assay （ELISA） to evaluate the ovarian reserve. The ovary and uterus indices were calculated. The serum levels of interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） were measured by enzyme-linked immunosorbent assay （ELISA）. The morphology of the ovarian tissue in the offspring on the day of birth and day 75 after birth was observed by hematoxylin-eosin staining. The transport of NF-κB p65 to the nucleus in the ovaries of the 75-day-old offspring was detected by the immunofluorescence （IF） assay. The expression of TLR4， NF-κB p65 and other related proteins in the ovarian tissue was determined by Western blot. ResultCompared with the control group， the model group showed reduced primordial follicles in the offspring on the day of birth （P<0.01） as well as disturbed estrous cycle， decreased ovary index and uterus index （P<0.01）， reduced corpus luteum， increased atretic follicles （P<0.01）， lowered serum levels of AMH and E2 （P<0.01）， elevated serum levels of LH， FSH， IL-1β， and TNF-α （P<0.05， P<0.01）， and up-regulated protein levels of TLR4， NF-κB p65， recombinant myeloid differentiation factor 88 （MyD88）， and phosphorylated NF-κB inhibitor （p-IκBα） （P<0.01） in the 75-day-old offspring rats. Compared with the model group， Zuoguiwan and vitamin E increased the primordial follicles in the offspring on the day of birth （P<0.01）. Moreover， they resumed the estrous cycle， increased the ovary and uterine indices （P<0.05， P<0.01） and corpus luteum （P<0.01）， reduced atretic follicles （P<0.01）， elevated the serum levels of AMH and E2 （P<0.05， P<0.01）， lowered the serum levels of LH， FSH， IL-1β， and TNF-α （P<0.05， P<0.01）， and down-regulated the expression of TLR4， NF-κB p65， MyD88， and p-IκB-α （P<0.05， P<0.01） in the 75-day-old offspring. ConclusionZuoguiwan can improve the ovarian reserve in the offspring rat model of congenital kidney deficiency by regulating the TLR4/NF-κB p65 signaling pathway.
ABSTRACT
Objective To investigate the different components of Acorus tatarinowii and Polygala tenuifolia(volatile oil, aqueous extract)on the expression of phosphorylated Tau protein at site Ser396 and Tau-5 in the hippocampus of rats with Alzheimer′s disease(AD). Methods Male Sprague Dawley rats were randomly divided into 8 groups:normal control group, model control group,low-,middle-,and high-dose groups of volatile oil of Acorus tatarinowii and Polygala tenuifolia,and low-, middle-,and high-dose groups of aqueous extract of Acorus tatarinowii and Polygala tenuifolia. The subacute aging model was established by intraperitoneal injection of D-galactose( D-gal). Rats were given different components of Acorus tatarinowii and Polygala tenuifolia(crude drug dosage,0.6,1.2,1.8 g·kg-1 )in experimental groups,and 0.9% sodium chloride solution in normal control group and model control group,by gavage for 28 days.The levels of phosphorylated Tau protein at site Ser396 and Tau-5 were detected in hippocampal tissues by Western blotting and immunohistochemistry. Results The levels of phosphorylated Tau protein at site Ser396 were significantly enhanced in the model control group,as compared with those in normal control group (P0.05). Conclusion Acorus tatarinowii and Polygala tenuifolia could promote the dephosphorylation of Ser396 site of Tau protein in the hippocampus of AD rats,with the aqueous extract component having better effects.
ABSTRACT
Objective:To study the antidepressant effect of Baihe Zhimu decoction (BZD)and its influence in the key factors (CaM,CaMKⅡ,CREB)of CaM signaling pathway in hippocampus of the rats with depression,and to explore the antidepressant effect of BZD. Methods:Fifty rats were divided into control group,model group, fluoxetine group,low and high doses of BDZ groups (n = 10).Expect for control group,all the rats in other groups were made depression models by means of chronic unpredictable mild stress along with isolated raising,for 21 d.Then the rats were fed with NS, fluoxetine (1.8 mg · kg-1 ), and BZD (1.5 and 3.0 g · kg-1 ), respectively;for 28 d.The learning and memory ability,autonomous activities and the fixed time in 5 min of the rats were tested by Morris water amaze,Open-field Test and Forced Swimming Test respectively. The damage and repair status of hippocampal neurons were observed by Nissl staining method;the expression levels of CaM,CaMKⅡ protein,CREB mRNA in hippocampus of the rats were detected by Western blotting and RT-PCR method. Results:Compared with model group,the total time of rats in the platform quadrant of Morris water maze in BZD groups and fluoxetine group,the total distance and the number of crossing platform were increased (P <0.05 or P <0.01),and the time of first crossing platform were shortened (P <0.01);the total scores in open field test were increased (P <0.01),the fixed time with 5 min in the forced swimming test was shortened (P <0.05 or P <0.01).Compared with fluoxetine group,the fixed time within 5 min of the rats in swimming test was shortened (P <0.05).The result of Nissl staining showed that the hippocampal neuron injury in BZD groups and fluoxetine group was improved compared with model group.The molecular test results showed that the CaM and CaMKⅡprotein expression levels in hippocampus of the rats in BZD groups and fluoxetine group were increased compared with model group (P < 0.05 or P < 0.01).Compared with model group,the CREB mRNA expression levels in fluoxetime group and BZD groups were increased (P < 0.05 or P < 0.01).Conclusion:BZD has antidepressant effect and can improve the hippocampal neuron injury of the rats with depression and its mechanism is related to increasing the expression levels of CaM,CaMKⅡ and CREB in hippocampus CAM signaling pathway of the rats.