| Objective：Type2diabetes (T2DM) and osteoporosis (OP) are bothcommon systemic metabolic diseases.The insidence of these two diseasesincreases year after year with the economic growth，changing of life style andaging of population. The trabecular bone density and fracture risk of thepatients of type2diabetes both increased. Researchs show that the PI3K-AKTpathway is the main insulin signal pathway and PI3, AKT-1, AKT-2and NFκBregulate bone metabolism by regulating the proliferation and differentiation ofbone and resisting apoptosis of bone. Skeletal muscle is not only an importantpart where the insulin resistance happens, but aslo a vital organ where thebone metabolism happens. The express of PI3K, AKT-1, AKT-2and NFκB inthe type2diabetic with osteoporosis skeletal muscle is unclear and there is noreport about whether they have common pathogenesis. In this study, weestablished the type2diabetes with osteoporosis rat model and measured theexpression of PI3K, AKT-1, AKT-2and NFκB in the skeletal muscle. Thisstudy aims to make a discussion about the expression level of PI3K, AKT-1,AKT-2and NFκB in type2diabetes osteoporosis rats skeletal musclepreliminary, and provide the theoretical foundation and the experimental basefor prevention and cure of type2diabetes osteoporosis.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 withhigh-sugar-fat feedstuff and injected small doses of STZ, and ovariectomied to make the osteoporosis rats model. At4weeks,8weeks,12weeks afterovariectomy, five rats from every group were randomly executed, thequadriceps femoris of which was separated after anesthesia. The expression ofPI3K, AKT-1, AKT-2and NFκB in protein level was detected by IHS(immunohistochemistry) while the expression of PI3K, AKT-1, AKT-2andNFκB in RNA level was detected by RT-PCR. The experimental data waspresented as mean±standard deviation. Variance Analysis was used to checkdifferences between groups by SPSS13.0，P<0.05was considered statisticallysignificant.Results：1Immunohistochemistry results: The expression of AKT-1, AKT-2, PI3Kand NFκB is located in the cytoplasm of the skeletal muscle cells. The chinogranules in the cytoplasm of the skeletal muscle cell indicate they aremasculine.1.1The expression of PI3K：at4week after ovariectomy，the expression ofPI3K in NOVX group is lower than that in NS group (P＜0.05), in DS groupis lower than that in NS group (P＜0.05), in DOVX group is lower than that inDS group (P＜0.05), and the expression of PI3K in DOVX group is lowerthan that in NOVX group (P＜0.05). At8week after ovariectomy, theexpression of PI3K in NOVX group is much lower than that in NS group (P＜0.05), in DS group is lower than that in NS group (P>0.05), in DOVX group issignificantly lower than that in DS and NOVX groups (P＜0.05). At12weekafter ovariectomy, the expression of PI3K in NOVX group is lower than thatin NS group (P＜0.05), in DS group is lower than that in NS group (P＜0.05),while in DOVX group is lower than that in DS (P＜0.05), in DOVX group islower than that in NOVX (P＜0.05).1.2The expression of AKT-1: at4week after ovariectomy, the expressionof AKT-1in NOVX group is lower than that in NS group (P＜0.05), in DSgroup is lower than that in NS group (P＜0.05), in DOVX group is lower thanthat in DS group (P＜0.05), in DOVX group is lower than that in NOVX group (P＜0.05). At8week after ovariectomy, the expression of AKT-1inNOVX group is slightly higher than in that NS group, but there is nosignificant difference between the two groups (P>0.05). The expression ofAKT-1in DS group is lower than that in NS group (P＜0.05)，in DOVX groupis lower than that in DS group,but there is no significant difference betweenthe two groups(P>0.05). The expression of AKT-1in DOVX group is lowerthan that in NOVX group (P＜0.05). At12week after ovariectomy, theexpression of AKT-1in NOVX group is lower than that in NS group (P＜0.05), in DS group is much lower than that in NS group (P＜0.05), in DOVXgroup is higher than that in DS group (P＜0.05), in DOVX group is lowerthan that in NOVX group, but there is no significant difference between thetwo groups(P>0.05).1.3The expression of AKT-2: at4week after ovariectomy, the expressionof AKT-2in NOVX group is lower than that in NS group (P＜0.05), in DSgroup is lower than that in NS group (P＜0.05), in DOVX group is lower thanthat in DS group (P＜0.05), and in DOVX group is lower than that in NOVXgroup (P＜0.05). At8week after ovariectomy, the expression of AKT-2inNOVX group is lower than that in NS group, but there is no significantdifference between the two groups (P>0.05), in DS group is lower than that inNS group(P<0.05), in DOVX group is slightly higher than that in DS group,but there is no significant difference between the two groups(P>0.05), inDOVX group is lower than that in NOVX group(P<0.05). At12week afterovariectomy, the expression of AKT-2in NOVX group is lower than that inNS group (P＜0.05), in DS group is lower than that in NS group (P＜0.05), inDOVX group is lower than that in DS group (P＜0.05), in DOVX group ismuch lower than that in NOVX group (P＜0.05).1.4The expression of NFκB: at4week after ovariectomy, the expressionof NFκB in NOVX group is higher than that in NS group (P＜0.05), in DSgroup is higher than that in NS group (P＜0.05), in DOVX group is higherthan that in DS group (P＜0.05), in DOVX group is much higher than that inNOVX group (P＜0.05). At8week after ovariectomy, the expression of NFκB in NOVX group is higher than that in NS group (P＜0.05), in DS groupis higher than that in NS group (P＜0.05), in DOVX group is higher than thatin DS group (P＜0.05), in DOVX group is higher than that in NOVX group (P＜0.05). At12week after ovariectomy, the expression of NFκB in NOVXgroup is higher than that in NS group, in DS group is higher than that in NSgroup, but there is no significant difference between the two groups (P>0.05),in DOVX group is higher than that in DS group (P＜0.05) and in DOVXgroup is also higher than that in NOVX group (P＜0.05).2RT-PCR results:2.1The expression of PI3K: at4week after ovariectomy, the expressionof PI3K in NOVX group is lower than that in NS group (P＜0.05), in DSgroup is much lower than that in NS group (P＜0.05), and in DOVX group islower than that in DS group, but there is no significant difference betweenthem(P>0.05), the expression of PI3K in DOVX group is lower than that inNOVX group (P＜0.05). At8week after ovariectomy,the expression of PI3Kin NOVX group is lower than that in NS group (P＜0.05), and in DS group islower than that in NS group (P<0.05), in DOVX group is lower than that inDS and NOVX group, but there is no significant difference between them(P>0.05). At12week after ovariectomy, the expression of PI3K in NOVXgroup is lower than that in NS group (P＜0.05), in DS group is lower than thatin NS group (P＜0.05), while in DOVX group is lower than that in DS group,but there is no significant difference between them (P>0.05). The expressionof PI3K in DOVX group is lower than that in NOVX group (P＜0.05).2.2The expression of AKT-1: at4week after ovariectomy, the expressionof AKT-1in NOVX group is lower than that in NS group(P＜0.05), in DSgroup is lower than that in NS group(P＜0.05), in DOVX group is lower thanthat in DS group, but there is no significant difference between them (P>0.05).The expression of AKT-1in DOVX group is lower than that in NOVX group(P<0.05). At8week after ovariectomy, the expression of AKT-1in NOVXgroup is lower than that in NS group (P＜0.05), in DS group is lower than that in NS group (P＜0.05), in DOVX group is lower than that in DS group, butthere is no significant difference between them (P>0.05). The expression ofAKT-1in DOVX group is lower than that in NOVX group (P<0.05). At12week after ovariectomy, the expression of AKT-1in NOVX group is lowerthan that in NS group, in DS group is lower than that in NS group, but thereare no significant difference between them (P>0.05). The expression of AKT-1in DOVX group is lower than that in DS group (P＜0.05), in DOVX group issignificantly lower than that in NOVX group (P＜0.05).2.3The expression of AKT-2: at4week after ovariectomy, the expressionof AKT-2in NOVX group is lower than that in NS group, in DOVX group islower than that in DS group, in DOVX group is lower than that in NOVXgroup, but there is no significant difference between them (P>0.05). Theexpression of AKT-2in DS group is lower than that in NS group(P＜0.05). At8week after ovariectomy, the expression of AKT-2in NOVX group is lowerthan that in NS group, in DS group is lower than that in NS group, in DOVXgroup is lower than that in DS group, in DOVX group is lower than that inNOVX group, but there is no significant difference between them (P>0.05). At12week after ovariectomy, the expression of AKT-2in NOVX group is lowerthan that in NS group (P＜0.05), in DS group is much lower than that in NSgroup (P＜0.05), in DOVX group is lower than that in DS group (P＜0.05), inDOVX group is much lower than that in NOVX group (P＜0.05).2.4The expression of NFκB: at4week after ovariectomy, the expressionof NFκB in NOVX and DS groups is both higher than that in NS group, butthere is no significant difference between them (P>0.05). The expression ofNFκB in DOVX group is higher than that in DS and NOVX groups(P<0.05).At8week after ovariectomy, the expression of NFκB in NOVX and DSgroups is both higher than that in NS group, in DOVX group is lower than thatin DS group，in DOVX group is higher than that in NOVX group, but there isno significant difference between them (P>0.05). At12week afterovariectomy, the expression of NFκB in NOVX and DS groups is both higher than that in NS group, in DOVX group is higher than that in DS and NOVXgroups, but there is no significant difference between them (P>0.05).Conclusion:1The expressions of PI3K, AKT-1and AKT-2in the skeletal muscle ofNOVX group is lower than that of NS group, while the expressions of NFκBof NOVX group is higher than that of NS group.2The expressions of PI3K, AKT-1and AKT-2in the skeletal muscle ofDS group is lower than that of NS group, while the expressions of NFκB ofDS group is higher than that of NS group.3The expressions of PI3K, AKT-1and AKT-2in the skeletal muscle ofDOVX group is lower than that of DC group, while the expressions of NFκBof DOVX group is higher than that of DS group.4The expressions of PI3K, AKT-1and AKT-2in the skeletal muscle ofDOVX group is lower than that of NOVX group, while the expressions ofNFκB of DOVX group is higher than that of NOVX group.5It may be relevant that the occurrence and development of type2diabetes with osteoporosis to the inhibition of the expressions of PI3K, AKT-1and AKT-2and the activation of the expressions of NFκB in the skeletalmuscle. |
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