The Effect Of Chinese Medicine For Tonifying Kidney Compound On The Related To The Regulation Of BMP2/RUNX2/MEK1Signal Networks In Postmenopausal Osteoporosisin Rats

Purpose: To observe the following ovariectomy-induced osteoporosis inpostmenopausal hypothalamus of rats，kidney，femur BMP2-related changes insignal transduction pathways using molecular biology techniques to explore themolecular biological mechanism of post-menopausal osteoporosis. Kidney ResearchCompound postmenopausal osteoporosis prevention mechanism for follow-upresearch complex kidney Side effects of postmenopausal osteoporosis andosteoporosis provide scientific basis for the theory of Chinese medicine in theprevention and treatment of postmenopausal clinical.Material and Method: SPF SD female rats10weeks old never mate.Randomization stratified by body weight，in addition to the normal group，sham operation group， the remaining rats were treated modeling both ovariesremoved. Sham group which only the lower back incision lines are not removedovarian tissue.7days after rats were given appropriate medication group wereintragastrically administered for12weeks. After gavage for12weeks， thecollected specimens. By dual energy X-ray absorptiometry left femur BMD wasmeasured. When drawn by electronic balance and body weight were measured inrats rat uterine wet weight，uterine index calculation.Using HE staining，light microscopy of rat femur pathological changes.Grouping and intervention methods are as follows: normal control group，sham operation group，blank group，low-dose group，dose group，high-dose group each group was given12weeks.Real Time-PCR was used to detect the hypothalamus of rats in each group， kidney，femur BMP2、SMAD1、RUNX2、OSX、MEK1、ERK2mRNA expression levels； use Westernblot method to detect the rat hypothalamus，kidney，femur BMP2、SMAD1、Runx2、OSX、MEK1、ERK2protein expression levels.Results:Experiment I:1Comparison of rats in each group left hind femur BMD from the body，comparedwith the normal control group，blank group femur BMD was significantly lowerthan the normal group (P＜0.01)，osteoporosis model successfully copied.12weeks after treatment，comparing with blank group significantly raised the levelof the femur bone mineral density (P＜0.05). Comparison between theexperimental groups，low and high dose group than Gaitianli group (P＜0.05)，but the low， between the high-dose group was not statistically significantdifferences.2Compares the rats uterus index，compared with the normal group，femur blankgroup was significantly lower than the normal group (P＜0.01)，osteoporosismodel successfully copied.12weeks after treatment，comparing with blank groupsignificantly raised the level of the femur bone mineral density (P＜0.05).Comparison between the experimental groups，low and high dose group thanGaitianli group (P＜0.05)， but the low，between the high-dose group was notstatistically significant differences.3HE staining was observed under light microscope right femur biopsy of the ratsin the control group of normal bone marrow cavity visible smaller trabecularthick， full morphological structure， visible on the surface of trabecular boneosteoblasts. Blank group marrow cavity increases trabecular bone fracture.Experiment II:The normal group，sham group，blank group BGP levels were significantly lower (P＜0.01)，illustrate successful modeling; kidney compound is low，after12weeks，the high-dose group and the group Gaitianli medication，comparisonwith the blank group significantly increased the BGP level (P＜0.01)，but nostatistically significant difference between groups; with the normal group，comparison，TRACP blank group level was significantly higher in sham group (P＜0.01)， kidney compound low，medium and high dose groups after treatment with12weeks Gaitianli group，and die empty group can significantly reduce the TRACPlevels (P＜0.01)，between the experimental group，the high-dose group than inthe low-dose group，the celestial dome force group (P＜0.01)，high-dose groupthan in the middle dose group (P＜0.05).Experiment III:1.Under the hypothalamus of rats, renal, femoral BMP2mRNA and protein levelsshowed that: The expression in the hypothalamus of the results, compared withthe normal group, the hypothalamus of rats BMP2mRNA blank group was decreased,but no significant difference (P＞0.05); medication after12weeks, comparedwith the blank group, the experimental group could cut its expression, butno significant difference (P＞0.05). Compared with the normal group, blankgroup BMP2protein expression levels were significantly increased (P＜0.01);compared with the blank group,low,medium and high dose groups in the hypothalamusBMP2protein expression levels were significantly lower (P＜0.01).The results showed that the expression in the kidney tissue: expressionin rat BMP2mRNA blank group compared with the normal group, reduce, but nosignificant difference (P＞0.05); medication after12weeks, compared withthe blank group, the high-dose group and Gaitianli groups can upregulate theexpression (P＜0.05) compared with the normal group, the lower blank group BMP2protein levels, but not statistically significant (P＞0.05); compared withthe blank group, low-dose group, high dose group BMP2elevated protein levels,but not statistically significant (P＞0.05)The results showed that the expression of the femur: Compared with the normal group, blank group of rats BMP2mRNA increased expression (P＜0.05); medicationafter12weeks, compared with the blank group, the experimental group couldcut its expression (P＜0.05). Compared with the normal group, BMP2proteinexpression level was significantly lower in rats blank group (P＜0.01); comparedwith the blank group,12weeks after treatment, low dose group BMP2proteinexpression levels were significantly increased (P＜0.01); compared with themiddle dose group and low dose group BMP2protein expression levels weresignificantly increased (P＜0.01).2.Under the hypothalamus of rats, kidney, femur Smad1mRNA and protein expressionanalysis showed that: the hypothalamus expression showed that expression ofrat Smad1mRNA blank group compared with the control group, lower (P＜0.05);12weeks after treatment,compared with the blank group, the experimental groupcould cut its expression,but no significant difference (P＞0.05). Comparedwith the normal group,blank group SMAD1protein expression levels weresignificantly increased (P＜0.01); compared with the blank group, low-dosegroup, middle dose group and high dose group, Gaitianli group in the hypothalamusSMAD1protein levels significantly lower (P＜0.01).The results showed that the expression in the kidney tissue: expressionin rat SMAD1mRNA blank group compared with the normal group, increased (P＜0.05); medication after12weeks, compared with the blank group, low-dose groupmay cut its expression (P＜0.05) compared with the normal group, blank groupSMAD1lower protein levels (P＜0.01); compared with the blank group, the high-dose group, Gaitianli group SMAD1expression levels were significantlyincreased (P＜0.01).The results showed that the expression of the femur: Compared with the normalgroup,rats SMAD1mRNA blank group was significantly decreased, but no significantdifference (P＞0.05); medication after12weeks, compared with the blank group,the high-dose group significantly raised its expression (P＜0.01). Comparedwith the normal group, SMAD1protein expression was significantly lower in ratsblank group (P＜0.01); compared with the blank group,12weeks after treatment, the low-dose group, middle dose group and high dose group, Gaitianli group SMAD1protein expression levels were significantly increased (P＜0.05); between theexperimental group, the low-dose group SMAD1protein expression levels weresignificantly increased (P＜0.01)Experiment IV:1Under the hypothalamus of rats, renal, femoral RUNX2mRNA and protein levelsshowed: hypothalamic expression results in the next, compared with the normalgroup, blank group RUNX2mRNA expression was significantly lower (P＜0.05);and blank group phase ratio, mRNA expression of RUNX2hypothalamus was increasedin low-dose group, but not statistically significant (P＞0.05) compared withthe normal group, significantly increased expression of Runx2blank group(P＜0.01);compared with the blank group low-dose group, middle dose group andhigh dose group, Gaitianli group Runx2protein expression was significantlylower (P＜0.01).Expression in renal tissue showed: Compared with the normal group, blankgroup RUNX2mRNA expression was significantly decreased, but not statisticallysignificant (P＞0.05); Compared with blank group, high dose group RUNX2mRNAexpression was significantly increased (P＜0.05) compared with the normalgroup, blank group Runx2protein expression was significantly lower (P＜0.01);compared with the blank group, middle dose group and low dose group and highdose group Gaitianli group Runx2protein expression, significantly reduced (P＜0.05).The results showed that the expression of the femur: Compared with the normalgroup, blank group RUNX2mRNA expression was significantly lower (P＜0.01);compared with the blank group, the high-dose group RUNX2mRNA expression wassignificantly increased (P＜0.01) compared with the normal group, blank groupRUNX2protein expression was significantly decreased, but not statisticallysignificant (P＞0.05); compared with the blank group, low-dose group, middledose group protein expression was significantly higher, but not statistically significant (P＞0.05).2．Under the hypothalamus of rats, kidney, femur OSX mRNA and protein levelsshowed: hypothalamic expression results in the next, compared with the normalgroup, blank group OSX mRNA expression was significantly lower (P＜0.05); andblank group phase ratio, mRNA expression in the hypothalamus OSX low dose groupwas increased, but not statistically significant (P＞0.05) compared with thenormal group, significantly lower protein expression OSX blank group (P＜0.01).Compared with the blank group low-dose group, middle dose group and high dosegroup, Gaitianli group OSX protein expression was significantly increased(P＜0.01).Expression in renal tissue showed: Compared with the normal group, blankgroup OSX mRNA expression was significantly increased (P＜0.05); Compared withblank group, low, medium and high dose groups OSX mRNA expression wassignificantly lower (P＜0.05) compared with the normal group, blank group OSXprotein expression was significantly increased (P＜0.01); compared with theblank group, middle dose group and high dose group, Gaitianli group OSX proteinexpression was significantly increased (P＜0.01).The results showed that the expression of the femur: Compared with the normalgroup, blank group OSX mRNA expression was significantly increased (P＜0.05);compared with the blank group,low,medium and high dose groups OSX mRNA expressionwas significantly lower (P＜0.05) and normal group,significantly higherexpression of OSX blank group (P＜0.01); compared with the blank group, low-dose group, middle dose group and high dose group, Gaitianli group OSX proteinexpression was significantly increased (P＜0.01).Experiment V:1Under the hypothalamus of rats, renal, femoral MEK1mRNA and protein levelsshowed: expression in the hypothalamus, compared with the normal group,MEK1mRNA expression blank group was significantly lower (P＜0.05); compared withthe blank group, low-dose group, MEK1mRNA express high dose group increased, but not statistically significant (P＞0.05) compared with the normal group,significantly higher expression of MEK1blank group (P＜0.01);compared with theblank group, low dose group, middle dose group and high dose group Gaitianligroup MEK1protein expression was significantly lower (P＜0.01).Expression in renal tissue showed: MEK1mRNA expression blank group comparedwith the normal group, increased, but not statistically significant (P＞0.05);Compared with blank group, MEK1mRNA express high-dose group, Gaitianli groupincreased.(P＜0.05) compared with the normal group, blank group MEK1proteinexpression was significantly lower (P＜0.01); compared with the blank group,low-dose group, high dose group MEK1protein expression was significantly lower(P＜0.05).The results showed that the expression of the femur: Compared with the normalgroup, MEK1mRNA expression blank group was significantly higher,but notstatistically significant (P＞0.05); Compared with the blank group, MEK1mRNAexpress high dose group was significantly lower (P＜0.05).Compared with thenormal group, blank group MEK1protein expression was significantly lower (P＜0.01); compared with the blank group, low-dose group, middle dose group andhigh dose group MEK1protein expression was significantly increased (P＜0.01).2.Under the hypothalamus of rats, kidney, femur ERK2mRNA and protein levelsshowed: expression in the hypothalamus, and the normal group, ERK2mRNAexpression blank group increased, but not statistically significant (P＞0.05);and compared, ERK2mRNA high dose group was significantly higher expression blankgroup (P＜0.01).Compared with the normal group, blank group ERK2proteinexpression was significantly increased (P＜0.01);Compared with the blank group,low-dose group, dose group, high dose group ERK2protein expression wassignificantly lower (P＜0.01); with low-dose group, middle dose group and highdose group ERK2protein expression was significantly lower (P＜0.05)Expression in renal tissue showed: ERK2mRNA expression blank group comparedwith the normal group, increased, but not statistically significant (P＞0.05);Compared with blank group, ERK2mRNA express high dose group increased (P＜ 0.01) compared with the normal group, blank group ERK2protein expression wassignificantly lower (P＜0.01); compared with the blank group, low-dose group,high dose group ERK2protein expression was significantly lower (P＜0.01).The results showed that the expression of the femur: Compared with the normalgroup, ERK2mRNA expression blank group was significantly lower (P＜0.01);compared with the blank group, low, ERK2mRNA expression dose increased, butnot statistically significant (P＞0.05) compared with the normal group, blankgroup ERK2protein expression was significantly increased (P＜0.01); comparedwith the blank group, low-dose group, middle dose group and high dose groupERK2protein expression was significantly increased (P＜0.01).Conclusion:1Under normal rat hypothalamus, kidney, femur express BMP2, SMAD1, RUNX2, OSX,MEK1, ERK2, showed normal rat hypothalamus, kidney, femur presenceBMP2/RUNX2/MEK1signal transduction pathway network.2Postmenopausal osteoporosis femur RUNX2, MEK1, reduces the expression levelERK2mRNA, BMP2, OSXmRNA elevated expression level, BMP2, SMAD1, MEK1expressionlevel decreasing OSX, ERK2protein levels; renal tissue BMP2, RUNX2, MEK1mRNAexpression decreased, increased SMAD1, OSXmRNA expression levels, Smad1Runx2,MEK1, ERK2protein expression, reduce, OSX protein expression increased;reduced hypothalamic SMAD1, RUNX2, OSX, MEK1mRNA expression levels, OSX proteinexpression decreased, BMP2, SMAD1, Runx2, MEK1, ERK2protein expressionincreased, is one of the pathogenesis of osteoporosis in postmenopausal women.3Kidney compound can significantly improve bone density in postmenopausalosteoporosis in rats.4Kidney compound can significantly increase serum levels of osteoporosis inpostmenopausal BGP rats,decreased serum levels of osteoporosis inpostmenopausal rat TRACP, and play a role in prevention and treatment ofosteoporosis in post-menopausal.5Kidney compound femur can BMP2postmenopausal osteoporosis in rats, OSX, MEK1mRNA expression levels were significantly lower elevated SMAD1, RUNX2mRNAexpression levels, BMP2, SMAD1, OSX, MEK1, ERK2protein expression increased;kidney tissue Smad1reduce OSXmRNA expression levels increased BMP2, RUNX2, MEK1,ERK2mRNA expression levels, reduce RUNX2, MEK1, ERK2protein levels, SMAD1,OSX protein expression increased; elevated hypothalamic ERK2mRNA expressionlevels, BMP2, SMAD1, RUNX2, MEK1, ERK2protein reduced, OSX protein expressionincreased; prompted by lower kidney compound may improve and regulate thehypothalamus, kidney, femur BMP2, SMAD1, RUNX2, OSX, MEK1ERK2of mRNA, proteinexpression,played prevention of postmenopausal bone interstitial osteoporosis.