| Objective: Osteoporosis, as a silent epidemic disease, gradually is beenrecognized. Numerous studies confirm that type1diabetes (T1DM) may leadto the occurrence of osteoporosis, but the relationship between type2diabetes(T2DM) and osteoporosis is not yet clear, studies suggest that insulinsignaling pathway may be related to them. PI3K-Akt signaling pathway is amajor downstream molecule of insulin pathway, its expression decreased inthe pathogenesis of diabetes and insulin resistance play an important role.NFκB signaling pathway can induce osteoclast differentiation, extendedsurvival time of osteoclasts and promote bone resorption. Study found thatPI3K-Akt signaling pathway control cell proliferation, apoptosis, glucosetransport and other biological activities by regulating NFκB, mTOR and otherdownstream factors. Insulin resistance occurs mainly in the kidney andskeletal muscle, but is often complicated by diabetic nephropathy withadvanced diabetes, and kidney take part in the incidence and process ofosteoporosis by regulating the balance between osteoblasts and osteoclasts.There is no report about PI3K,Akt1,Akt2,NFκB expression in the kidney ofType2diabetic osteoporosis Rats.The aim of this study is to explore theexpression of PI3K, Akt1, Akt2and NFκB in the kidney of Type2diabeticosteoporosis Rats, in order to understand the mechanism of diabeticosteoporosis and provide experimental and theoretical basis.Methods:100Wistar female rats, only2.5to3months, were randomlydivided into four groups after1week feeding: normal control group(shamgroup, NS,N=24), ovariectomy group(NOVX,N=26), type2diabetes controlgroup(sham group, DS,N=24), type2diabetic rats under ovariectomy group(DOVX,N=24). The weight of rats in the four groups had no significantdifference.The T2DM rats model was made by feeding them with high-sugar-fat feedstuff and injected small doses of STZ, and ovariectomied tomake the osteoporosis rats model.At4weeks,8weeks,12weeks afterovariectomy, five rats from every group were randomly executed, the kidneyof which was separated after anesthesia. Renal tissue RNA was extrated fromthe kidney. The expression of PI3K, Akt1,Akt2and NFκB in protein level wasdetected by IHS (immunohistochemistry) while the expression of PI3K,Akt1,Akt2and NFκB in mRNA level was detected by RT-PCR. SPSS13.0statistic software was used to process the experimental data.The experimentaldata was presented as mean±standard deviation. Variance Analysis was usedto check differences between groups, P<0.05was considered statisticallysignificant.Results:1The pathology of the kidney: Under microscope, the glomerular ofnon-diabetic groups is clear,the capillary loops of glomerular is thin andclear.The number of Endothelial cells and mesangial cells is normal.There isno significantly difference between NS group and NOVX group. In DS andDOVX groups, eosinophilic material in renal glomerular capillariesincreased.The number of endothelial cells and mesangial cells also increasedin DS and DOVX groups.2The expression level of PI3K2.1Immunohistochemistry results: PI3K mainly expressed in nucleus andcytoplasm of the renal tubular epithelial cells. At4,8weeks after ovariectomy,the expression in DOVX and DS groups is significantly lower than that in NSgroups (P<0.05), but there is no difference between NOVX and NS group(P>0.05). At12weeks after ovariectomy, the expression in NOVX,DS andDOVX groups is much lower than that in NS group (P<0.05), there is nosignificantly difference between DS and DOVX groups(P>0.05).2.2RT-PCR results: At4weeks after ovariectomy, the expression in NOVXgroup is much higher than taht in NS group(P<0.05), in DOVX group ismuch lower than that in DS group(P<0.05), the expression in DS and DOVXgroups is much higher than that in NS and NOVX groups respectively(P＜ 0.05).At8weeks after ovariectomy, the expression in NOVX groups is muchlower than that in NS group(P<0.05), in DOVX group is much higher than thatin DS and NOVX groups(P<0.05), in DS group is much lower than that in NSgroup(P＜0.05).At12weeks after ovariectomy, the expression in NOVXgroups is much higher than that in NS group(P<0.05), in DOVX group ismuch lower than that in DS and NOVX groups(P<0.05), in DS group is muchhigher than that in NS group(P＜0.05).3The expression level of Akt13.1Immunohistochemistry results: Akt1expressed in cytoplasm of the renaltubular epithelial cells. At4,8weeks after ovariectomy, the expression of Akt1in DOVX and DS groups is significantly lower than that in NS group(P<0.05),there is no difference between NOVX and NS group(P>0.05); at12weeksafter ovariectomy, the expression of Akt1is no significantly difference in fourgroups(P>0.05).3.2RT-PCR results: At4weeks after ovariectomy, the expression of Akt1inDOVX group is significantly lower than that in DS(P<0.05),in DS group issignificantly higher than that in NS group(P<0.05), the expression of Akt1isno significantly differences between NOVX and NS groups, NOVX andDOVX groups(P>0.05).At8weeks after ovariectomy, the expression inNOVX group is significantly higher than that in NS(P<0.05), there is nosignificantly differences between DOVX and DS groups. The expression ofAkt1in DS group is much lower than that in NS group(P<0.05), in DOVXgroup is much lower than that in NOVX group(P<0.05).At12weeks afterovariectomy, the expression in NOVX group is significantly higher than thatin NS group(P<0.05), in DOVX group is lower than that in DS group(P<0.05),in DS group is lower than that in NS group(P<0.05), in DOVX group is lowerthan that in NOVX group(P<0.05).4The expression level of Akt24.1Immunohistochemistry results: Akt2mainly expressed in cytoplasm of therenal tubular epithelial cells. At4weeks after ovariectomy, the expression ofAkt2in DOVX,DS and NOVX group is significantly lower than that in NS group(P<0.05).At8weeks after ovariectomy, the expression in DS group ismuch lower than that in DOVX, NS, NOVX groups(P<0.05), the expressionof Akt2in DOVX, NOVX group is significantly lower than that in NS group(P＜0.05).At12weeks after ovariectomy, the expression in DOVX,DS andNOVX group is much lower than that in NS groups(P<0.05).4.2RT-PCR results: At4weeks after ovariectomy, the expression of Akt2inNOVX group is significantly higher than that in NS group(P<0.05), in DOVXgroup is much lower than that in DS group(P<0.05),there is no significantlydifferences between NS and DS group(P>0.05). The expression of Akt2inDOVX group is much lower than that in NOVX group(P<0.05). At8weeksafter ovariectomy, the expression of Akt2is no significantly differences infour groups(P>0.05). At12weeks after ovariectomy, the expression in NOVXgroup is significantly higher than that in NS, in DOVX group is much lowerthan that in DS group (P<0.05), in DS group is much lower than that in NSgroup (P<0.05), in DOVX group is much lower than that in NOVX group(P<0.05).5The expression level of NFκB5.1Immunohistochemistry results: NFκB expressed in cytoplasm of the renaltubular epithelial cells.At4,8weeks after ovariectomy, the expression level ofNFκB is no significantly differences between NS,NOVX,DS and DOVXgroups(P>0.05).At12weeks after ovariectomy, the expression in DS group ismuch lower than that in NS.The expression level of NFκB is no significantlydifferences between DOVX, NOVX groups and DS,NSgroups(P>0.05).5.2RT-PCR results: At4weeks after ovariectomy, the expression of NFκB inNOVX group is significantly higher than that in NS group(P<0.05), in DSgroup is much higher than that in NS group(P<0.05), the expression of NFκBis no significantly differences between DOVX and DS group,DOVX andNOVX group(P>0.05). At8weeks after ovariectomy, the expression of NFκBis no significantly differences between NOVX and NS groups (P>0.05), theexpression in DOVX group is significantly lower than that in DS (P<0.05),in DS group is much lower than that in NS group (P<0.05), in DOVX group is much lower than that in NOVX group (P<0.05).At12weeks after ovariectomy,the expression in NOVX group is significantly higher than that in NS (P<0.05),in DOVX group is much lower than that in DS group (P<0.05), in DS group ismuch higher than that in NS group (P<0.05), in DOVX group is much lowerthan that in NOVX group (P<0.05).Conclusion:1The expression of PI3K, Akt1,Akt2and NFκB in kidney of NOVX grouprats is lower than that of NS group rats.2The expressions of PI3K, Akt1,Akt2and NFκB in kidney of DS group ratsis lower than that of NS group rats.3The expressions of PI3K, Akt1,Akt2and NFκB in kidney of DOVX grouprats is lower than that of NS,NOVX and DS group rats.4The inhibition of the inslin signaling expressions of PI3K,Akt, NFκB inkidney pathway maybe related to the pathogenesis of Type2diabetes withosteroposis. |
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