| ObjectiveIn this study,D-galactose-induced aging model rats were used as experimental subjects,and the effects of Ganoderma lucidum Rhodiola compound preparation(GRCP)on cognitive function,anti-oxidative stress and immune function of aging rats were investigated.The aim was to evaluate the anti-aging effects of GRCP.MethodsForty-eight SPF male SD rats were randomly divided into six groups:normal control(NC)group,D-galactose(D-gal)group,positive control(D-gal+VC)group,low-dose(D-gal+LCP)group,middle-dose(D-gal+MCP)group and high-dose(D-gal+HCP)group.Rats in the NC group were subcutaneously injected with saline every day,and distilled water was given.Rats in other groups were injected subcutaneously with D-galactose at a dose of 400mg/kg daily to establish the aging model.At the same time,the rats in the D-gal+VC group were administrated with 100mg/kg vitamin C,and the drug group was given 25mg/kg,50mg/kg and 100mg/kg GRCP.Dosing was continued for 8 weeks,and body weight was measured once a week.The Morris Water Maze experiment was used to calculate the behavior of rats from jumping into the water to escaping from the platform to test their behavioral changes.At the end of the experiment,the animals were killed after anesthesia,serum,tissues and organs were collected.The spleen and thymus of rats were weighed to measure the spleen index and thymus index.Prepare a spleen cell suspension and stimulate the proliferation of T lymphocytes and B lymphocytes with 2μg/mL ConA and 10μg/mL LPS.MTS/PMS method was used to detect the proliferation ability of T cells and B cells.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of IL-1β,IL-2,IL-4,IL-6,lipofuscin,TNF-α and TGF-β in rats.Rat serum was collected to measure anti-oxidation related indexes.The rat hippocampus was isolated,and the levels of MyD88 and NF-κB p65 protein were detected by Western Blot.Results1.Compared with the rats in D-gal group,the weight of the rats in the D-gal+HCP group increased significantly(P<0.05).Compared with D-gal group,rats in the other groups did not alter significantly body weight(P>0.05).The results of the Morris Water Maze experiment showed that,compared with the NC group,rats in the D-gal group took a longer time to find the hidden platform(P<0.01).Compared with aging rats,the time required for rats in the VC group,MCP group,and HCP group was reduced(P<0.05).2.The spleen and thymus index were decreased in the D-gal group(P<0.01).The spleen and thymus index were rebounded by VC and GRCP treatment(P<0.05).In addition,compared with the NC group,D-gal inhibited the proliferation of T and B lymphocytes of splenocytes(P<0.01).The proliferative ability of T and B lymphocytes in the drug group was higher than that in the D-gal group(P>0.05).Furthermore,D-gal+HCP facilitated faster B and T lymphocyte proliferation of splenocyte(P<0.01,P<0.05).3.Compared with the NC group,the inflammatory factors IL-1β,IL-6 and TNF-α in the aging rats were significantly increased(P<0.01).The D-gal+VC group showed significant improvement(P<0.05).The inflammatory factors of D-gal+HCP group and MCP decreased significantly(P<0.01,P<0.05),and the level of IL-6 in D-gal+LCP group also decreased(P<0.01).4.Compared with the NC group rats,the IL-2 and IL-4 levels in the D-gal group was significantly reduced(P<0.01),and the level of the rats in the VC group was significantly increased(P<0.01).Rats in the D-gal+HCP group had the most pronounced effect(P<0.05).Compared with the NC group,the Lipo level in the D-gal group was significantly increased(P<0.01),and the VC group,LCP group,MCP group,and HCP group were significantly improved(P<0.01).The TGF-β level in the D-gal group was significantly increased(P<0.01),the level in the VC group was significantly reduced(P<0.01),there was no significant statistical difference between the LCP and MCP groups(P>0.05),and the HCP group was significantly reduced(P<0.01).5.Compared with the NC group,the T-AOC,SOD,GSH-PX,and CAT activities in the serum of the D-gal group rats were significantly reduced(P<0.01).Compared with D-gal group,VC group,MCP and HCP group were significantly improved(P<0.01,P<0.05).The levels of T-AOC,SOD,and CAT in the LCP group rats were also significantly increased(P<0.05).Compared with the NC group,the MDA activity in the serum of the D-gal group was significantly increased(P<0.01).Compared with D-gal group,VC group and HCP group were significantly improved(P<0.01).6.The expression of MyD88 and NF-κB p65 in hippocampus were significantly increased in D-gal group,compared to the NC group(P<0.01).Moreover,a regulatory role of GRCP treatment on MyD88 and NF-κB expression in rat hippocampus was also observed.VC and GRCP administration reduced MyD88 and NF-κB p65 levels,while MyD88 and NF-κB p65 expression was also decreased dramatically in D-gal+HCP group.(P<0.05).ConclusionGRCP has significant anti-aging activity in rats.Rats in the D-gal+HCP group has the best anti-aging activity.GRCP can improve the cognitive function of senescent rats,inhibit the oxidative stress in senescent rats,enhance the immune function of senescent rats,and reduce the secretion of inflammatory factors by reducing the expression of MyD88 protein in the hippocampus and blocking the NF-κB signaling pathway in senescent rats.This research provides a theoretical basis for the development of related health foods. |
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