| Purpose: This topic,in view of TCM viscera-state theory,that "the kidney stores up essence and bears marrow and grew bone up,according to the theory of mutual aid of Yin and Yang" selects TCM classic prescription of invigorating the kidney and strengthening the essence,Zuogui pill and Yougui pill.By studying Zuogui pil and Yougui pill to ovariectomized ratsbone marrow stromal mesenchymal cells in the process of osteogenesis,into fat balance the relationship between apoptosis and to explore the prevention and control of left and right to pill,the mechanism of postmenopausal osteoporosis to TCM provide certain experiment basis for prevention and treatment of postmenopausal osteoporosis.Material and method : Sixty Sprague-Dawley female 2-momth-old 60 rats were randomly divided into 6 groups: Control group(Control),Ovariectomy group(OVX)Sham-operated group(Sham),Zuogui pill(ZGW),Yougui Pill(YGW)and prognova group.By bilateral ovariectomy,establish postmenopausal osteoporosis model.The rats were given 6.3times dose of human’s dose 3d after gavage feeding,and once a day.For 12 weeks,use whole bone marrow attachment method to get the BMSCs of ovariectomized rats and culture in vitro.By intervening the BMSCs of ovariectomized rats and using flow cytometry to identify BMSCs,and MTT assay was used to detect the proliferation of BMSCs.BMSCs were induced by osteogenesis and adipogenesis,the alkaline phosphatase activity of each group was detected by p NPP and ELISA.The expression of ALP was observed by modified calcium cobalt staining method.The formation of intracellular mineralized nodules was observed by alizarin red staining.The expressions of Collagen I(COLI)and Runt-related transcription factor 2(Runx 2)gene and protein were detected by RT-q PCR and Western blot.The formation of lipid droplets in the cells was observed by oil red O staining.The expressions of Peroxisome proliferator-actic receptor(PPARγ)and lipoprolipo protein lipase(LPL)gene and protein were detected by RT-q PCR and Wesrern blot,too.In the model group,10-6 mol · L-1,10-5mol · L-1,5×10-5mol · L-1 dexamethasone(Dex)were added for 24,48 and 72 h,respectively the apoptotic rate was measured by flow cytometry.The apoptosis rate of BMSCs in BMSCs treated with 10-6 mol · L-1Dex for 48 h was measured by flow cytometry,and the apoptotic rate of BMSCs after osteogenesis and adipogenic differentiation was compared.The expression of caspase-3,Bcl-2 were detected by RT-q PCR and Wesrern blot.After apoptosis,the expression of Runx2,COLI,PPARγ and LPL gene and protein.MTT assay was used to detect the proliferation of osteoblasts h FOB1.19 at different concentrations,then,the apoptotic rate of h FOB1.19 cells induced by Dex was detected by Annexin V / PI double staining method,detected the different concentrations of Verbascoside apoptosis.The apoptosis of h FOB1.19 cells was detected by Hoechest33258 and TUNEL.We detected the m RNA of Bcl-2 and Bax by the method of RT-q PCR,and detected protien level of Casepase-3、Bcl-2 and Bax by the method of Westrern blotting.The expression of ERK,p-ERK and Bcl-2 protein was detected by Wesrern blot method using ERK1 / 2 signal blocker method to treat h FOB1.19 with PD98059(ERK1 / 2 specific blocker).Results:1 The study of Zuogui pill and Yougui pill on ovarian rats proliferation of BMSCs1.1 The identification about BMSCs The extracted cells were cultured to P3 generation,using flow cytometry to identify three kinds of cell surface factor CD29,CD90,CD11b/c,and CD29 and CD90 of them were positive expression,the positive expression rate: 99.9%,98.7%.And CD11b/c was the negative expression,the negative expression rate is 14.3%.It is commonly determined that the extracted cells were BMSCs in this experiment,combined with the observation of cellullar morphology map and the result of cell flow identification.1.2 BMSCs growth curve and Zuogui pill and Yougui pill to the proliferous affection of ovaniectomized rats BMSCs1.2.1 BMSCs growth curve The results showed that BMSCs` growth curve was nearly S shape with the inoculum density increased and the optical density value enlarged.The incubation period was the front3d after cell inoculation,On the 4-th day,cells began to enter the logarithmic phase,and the growth of the cells reached the peak on the 7-th day.1.2.2 Zuogui,Yougui pill affect to ovarian rats proliferation of BMSCs The results showed that,compared with the control group,the cells proliferous rate in the model group were clearly reduced,the sham operation group and blank group had no significant difference,and the cell proliferation rate was close to the blank group.In the other three groups,medication administered,Zuogui pill to promote the proliferation of ovariectomized rats BMSCs was the best.2 The study on Zuogui pill and Yougui pill to the osteogenesis and adipogenic differentiation of ovariectomied rats BMSCs2.1 Zuogui pill and Yougui pill to the affection of ovariectomied rats BMSCs ALP2.1.1 Modified calcium cobalt staining method The results showed that,compared with the control group,the sham operation group had no significant change.In the other four groups,brown granules were decreasedin the microscope,in the model group brown granules were least;compared with the model group,in the blank group,the sham operation group and the other groups medication administered,brown granules were significantly increased,observed in the microscope.ALP expression remarkable enhanced;and compared with Zuoguipill group,the BJL group did not change significantly,in the Youguipill group brown granules reduced;and it is stated that Zuoguipill can enhance ALP expression after BMSCs` osteogenic differentiation.2.1.2 p NPP method The results showed that,compared with the control group,in each period of time,ALP activity in the model group was significantly decreased(P<0.05);and ALP activity in the other groups all rose in the groups medication administered,Zuoguipill group and the BJL group,in different periods,can enchance the ovariectomied rats BMSCs ALP activity increased,in Zuoguipill group,the effection was better.2.1.3 ELISA method The results showed that,compared with the control group,in the model group,the ALPcontent was significantly decreased(P<0.01),in Zuogui pill,Yougui pill and the BJL group,ALP content was increased;compared with the model group,in the other group,ALP contents were significantly increased(P<0.05);compared with the BJL group,in Yougui pill group,decreased significantly(P<0.05);comparison of the BJL group,there was no significant difference with Zuogui pill group,in Youguipill group,the content of ALP was significantly decreased(P<0.05).In all the medication groups,Zuoguipill group,ALP content was the highest.2.2 Zuogui pill,Yougui pill on the influence of the ovariectomized rats BMSCs mineralized nodule formation The results showed that,compared with the control group,there was no significant difference in the shame operation group.In the OVX and YGW,mineralized nodules significantly decreased;compared with the model group,in the shame operation group and the other medication groups,red brown mineralized nodules significantly increased,observed in the microscope;compared with the BJL group and Zuoguipill group,in Youguipill group;the mineralized nodules showed reduced.It makes clear that Zuoguipill can enhance the formation of mineralized nodules after the osteogenic induction of BMSCs.2.3 Affect of Zuogui pill,Yougui pill on the affection of ovariectomized rats BMSCs COL I and Runx2 m RNA and protein expression m RNA testing results showed that,compared with the control group,there was no significant difference in the shame operation group(P>0.05),in Zuoguipill group,Yougui pill group and the BJLgroup,the expression level of BMSCs COLI and Runx2’s m RNA decreased significantly(P<0.05);compared with the BJLgroup,the expression level of BMSCs COLI and Runx2 m RNA significantly increased in Zuoguipill group(P<0.05),compared with Zuoguipill group,in Youguipill group,the expression level of BMSCs COLI and Runx2 m RNA decreased significantly(P<0.05).Protein testing results showed that,compared with the control group,there was no significant difference in the shame operation group(P>0.05),in the model group and the other medication groups,the protein expression level of BMSCs COLI and Runx2significantly lowered(P<0.05);compared with the OVX,in Zuoguipill group and the BJL group,the protein level of BMSCs COLI and Runx2 significantly rose(P<0.05);compared with the BJL group,in Yougui pill group,the expression level of BMSCs COLI and Runx2 protein decreased significantly(P<0.05);and compared with Zuoguipill group,in Youguipill group,the expression levels of COLI and Runx2 protein significantly decreased(P<0.05).2.4 Zuogui pill,Yougui pill to the effect of lipid droplet formation of ovariectomized rats BMSCs The results showed that,compared with the control group,in the model group and Yougui pill group,the number of lipid droplets increased;and compared with the model group,in the other 4 groups observed,the number of lipid droplets decreased significantly;and compared with the BJL group,in Yougui pill group the number of lipid droplets increased;compared with Zuogui pill group,in Yougui pill group the number obviously increased;it is stated that Zuoguipill can inhibit the formation of ovariectomized rats BMSCs `adipocytes in adipogenic differentiation and Yougui pill can promote the formation of lipid droplets.2.5 Zuogui pill,Yougui pill on the affects of the m RNA and proteinexpression of ovariectomized rats BMSCs PPARγ and LPL m RNA testing results showed that,compared with the control group,in the model group,Youguipill group and the BJL group,the expression of BMSCs PPARγ m RNA significantly enhanced(P<0.01);the expression levels of BMSCs LPL m RNA in the model group significantly increased(P<0.01),the expression levels of PPARγ m RNA in Zuoguipill group decreased significantly(P<0.01);and the expression levels of LPL m RNA in Yougui pill were significantly lowered(P<0.05).And compared with Zuoguipill group,the expression levels of BMSCs PPARγ m RNA significantly rose(P<0.01);the expression levels of LPL m RNA in Youguipill group were significantly reduced(P<0.05).Protein test results show that,compared with the control group,in the model group,Youguipill group and the BJL group,the expression of BMSCs PPARγ protein significantly increased(P<0.01);the expression of BMSCs LPL protein in the model group increased significantly(P<0.01),the expression level of LPL protein decreased obviously inYougui pill group(P<0.05);and compared with the model group,in Zuogui pill group,after adipogenic differentiation the expression of BMSCs PPARγ protein decreased significantly(P<0.01),the expression of BMSCs LPL protein in Zuogui pill,Yougui pill group decreased significantly(P<0.01);and compared with the BJL group,in Zuogui pill group,the expression of PPARγ protein significantly decreased(P<0.05),the expression of PPARγ proteinin Yougui pill group significantly rose(P<0.01);the expression of LPL protein in Yougui pill group was down,but no significant difference;and compared with Zuogu pill group,the expression of BMSCs PPARγ protein in Yougui pill group significantly increased(P<0.01);the expression level of LPL protein was significantly decreased in Yougui pill group(P<0.05).3 The study on Zuogui pill,Yougui pill to the osteogenesis and adipogenic apoptosis of ovariectomized rats BMSCs3.1 The study on ovariectomized rats BMSCs`apoptosisby by dexamethasone Using Annexin V/PI double staining method,the ovariectomized rats BMSCs were carried through apoptosis test by different time and different concentration by dexamethasone inducing.Through flow diagram,the flow rate of the right lower quadrant and right upper quadrant was compared with,the results showed that the concentration of 10-5 mol · L-1dexamethasone 48 h was the optimum conditions of induced apoptosis.3.2 Zuogui pill,Yougui pill to the affection of the osteoblast apoptosis of ovariectomized rats BMSCs3.2.1 Zuogui pill,Yougui pill on the effect of ovariectomized rats BMSCs flowapoptosis The result of Annexin V/PI double staining method showed that,compared with the control group,in the model group and Youguipill group BMSCs apoptosis rate after bone-formation increased significantly(P<0.05);compared with OVX,the shame operation group and 3 medication groups,the apoptosis rate after BMSCs osteogenesis decreased(P<0.05);compared with the BJL group,in Zuogui pill group,the apoptosis rate decreased significantly(P<0.05);and compared with Zuogui pill group,significantly increased the apoptosis rate in Yougui pill group(P<0.05).The comparison with the apoptotic rate of groups before and after osteogenesis,in the BJL group,the apoptosis rate after osteogenesisincreased(P<0.05),in the other groups,the apoptosis rate after osteogenesis all significantly reduced(P<0.05.3.2.2 Zuogui pill,Yougui pill on the affect of the expression of ovariectomized rats BMSCs Caspase-3 m RNA and prepotein m RNA test results showed that,compared with the control group,the expression level of BMSCs Caspase-3 m RNA in the model group was significantly increased(P<0.05),the expression level of BMSCs Caspase-3 m RNA in Yougui pill group was significantly decreased(P<0.01);compared with the model group,in the shame operation group and the medication groups,the expression level of BMSCs Caspase-3 m RNA was significantly lowered(P<0.05);and compared with the BJL group,in Zuogui pill group the expression level of Caspase-3 m RNA was significantly lowered(P<0.05);and compared with Zuogui pill group,the expression level of BMSCs Caspase-3 increased significantly in Youguipill group(P<0.05).Protein testing results showed that,compared with the control group,the expression level of BMSCs Caspase-3 proteinin the model group was significantly increased(P<0.05),and in Zuoguipill,Youguipill group the expression of BMSCs Caspase-3 protein decreased significantly(P<0.05);and compared with the model group,in the other 4 groups,the expressionof BMSCs Caspase-3 protein all were significantly decreased(P<0.05)and compared with the BJL group,Zuogui pill ’ s Caspase-3 protein expression decreased significantly(P<0.05);and compared with Zuogui pill group,in Yougui pill group,significantly increased the expression of Caspase-3 protein(P<0.05).3.2.3 Zuogui pill,Yougui pill on the affect of the expression Bcl-2 m RNA and protein of ovariectomized rats BMSCs m RNA testing results showed that,compared with the control group,the expression level of BMSCs Bcl-2 m RNA in the model group and Yougui pill group decreased significantly(P<0.01),the expression level of BMSCs Bcl-2 m RNA in Zuoguipill group were significantly increased(P<0.01);and compared with the model group,in the shame operation group,Zuogui pill group and the BJL group,the expression level of BMSCs Bcl-2 m RNA increasedsignificantly(P<0.01);compared with the BJL group,in Zuogui pill group,the expression level of Bcl-2 m RNA was significantly increased(P<0.01),in Yougui pill group,the expression was significantly lower(P<0.05)and compared with Zuogui pill,in Yougui pill group and the BJL group,the expression level of BMSCs Bcl-2 m RNA decreased significantly(P<0.01).Protein testing results showed that,compared with the control group,in the other 4groups,the expressionlevel of BMSCs Bcl-2 protein was significantly decreased(P<0.05);and compared with the model group,in the medication group,the expression level of BMSCs Bcl-2 protein was significantly increased(P<0.05);and compared with the BJL group,in Yougui pill group,Bcl-2 protein level was significantly decreased(P<0.05),and compared with Zuogui pill group,BMSCs Bcl-2 protein level in Yougui pill group was significantly decreased(P<0.05).3.2.4 Zuogui pill,Yougui pill on the effect of the expression m RNA and protein of ovariectomized rats BMSCs COLI and Runx 2m RNA testing results showed that,compared with the control group,in the model group and Zuoguipill group,BMSCs COLI and Runx2 m RNA`s expression had significant difference(P<0.01),the expression in the model group was significantly decreased,the expression in Zuogui pill group increased significantly;and compared with the model group,in the shame operation group and the medication group,BMSCs of COLI and Runx2 m RNA`s expression increased significantly(P<0.01);and compared with the BJL group,in Zuogui pill group COLI and Runx2 m RNA`s expression was significantly increased(P<0.05);and compared with Zuogui pill group,in Yougui pill group and the BJL group,the expression of BMSCs COLI and Runx2 m RNA decreased significantly(P<0.05).Protein testing results showed that,compared with the control group,the expression level of BMSCs COLI and Runx2 proteinin in the model group decreased but in Zuogui pill group,the protein expression increased(P<0.05);and compared with the model group,in the shame operation group and medication group,BMSCs COLI and Runx2 protein`s expression was significantly increased(P<0.05);and compared with the BJL group,in Zuogui pill group,COLI and Runx2 protein`s expression was significantly increased(P<0.05);and compared with Zuoguipill group,in Yougui pill group and the BJL group,the expression of BMSCs COLI and Runx2 protein decreased significantly(P<0.05).3.3 Zuogui pill,Yougui pill on the effect about the adipogenic apoptosis of ovariectomized rats BMSCs3.3.1 Zuogui pill,Yougui pill on the effect to the flow type apoptosis of ovariectomized rats BMSCs The results of Annexin V/PI double staining method showed that,compared with the control group,in the model group,Zuoguipill group and the BJL group significantly increased the apoptosis rate of BMSCs(P<0.05);and compared with the model group,in the nedication group,the apoptosis rate of BMSCs decreased,there was significant difference(P<0.05);and compared with the BJL group,in Yougui pill group the apoptosis rate significantly decreased(P<0.05);and compared with Zuogui pill group,BMSCs` apoptosis rate outstandingly decreased in Yougui pill group(P<0.05).The comparison of the apoptotic rate in groups,before and after the adipogenesis,in addition to Yougui pill group,the rest of groups were increased.Among them,the model group and Zuogui pill group increased significantly,in Yougui pill group reduced the apoptosis rate.3.3.2 Zuogui pill,Yougui pill on the effect to the expression m RNA and protein of ovariectomized rats BMSCs Caspase-3m RNA testing results showed that,compared with the control group,in the model group,Zuogui pill group and the BJL group significantly increased the expression m RNA of BMSCs Caspase-3(P<0.01);compared with the model group,in the medication group,the expression of BMSCs Caspase-3 m RNA all decreased(P<0.05);and compared with the BJL group,the expression of m RNA in Yougui pill group decreased but there was no significant difference.(P>0.05);and compared with Zuogui pill group,the expression of Caspase-3 m RNA decreased in Yougui pill group,but no significant difference(P>0.05).Protein testing results showed that,compared with the control group,except for the shame operation group,in the 4 groups,BMSCs Caspase-3 protein`s expression levels weresignificantly increased(P<0.05);and compared with the model group,in the 3 medication group,the expression level of BMSCs Caspase-3 protein was significantly lowered(P<0.05);and compared with Zuogui pill group,in Yougui pill group,the expression level of BMSCs Caspase-3 protein reduced(P<0.05).3.3.3 Zuogui pill,Yougui pill on the effect to the expression m RNA and protein of ovariectomized rats BMSCs Bcl-2m RNA testing results showed that,compared with the control group,in the model group and Zuogui pill group,the expression of BMSCs Bcl-2 m RNA decreased significantly(P<0.05);and in Yougui pill group significantly increased the expression of BMSCs Bcl-2m RNA(P<0.05);and compared with the model group,in the other 4 groups,Bcl-2 m RNA`s expression was significantly increased(P< 0.05);and compared with the BJL group,the expression of Bcl-2 m RNA in Yougui pill group increased significantly(P<0.01);and compared with Zuogui pill group,in Yougui pill group significantly increased the expression of BMSCs Bcl-2 m RNA(P<0.01).Protein test results showed that,compared with the control group,except for the sham operation group,in the other 4 groups the expression level of BMSCs Bcl-2 protein decreased significantly(P<0.01);and compared with the model group,in the shame operation group and the other 3 groups,the expression level of BMSCs Bcl-2 protein was significantly increased(P<0.05);and compared with the BJL group,in Yougui pill group significantly increased the expression of Bcl-2 protein(P<0.05);and compared with Zuogui pill group,the expression of BMSCs Bcl-2 protein in Yougui pill group increased significantly(P<0.05).3.3.4 Zuogui pill,Yougui pill on the effect to the expression m RNA and protein of ovariectomized rats BMSCs PPARγm RNA test results showed that,compared with the control group,the expression of BMSCs PPARγ m RNA in the OVX increased significantly(P<0.05),in the medication group,the expression of BMSCs PPARγ m RNA all decreased(P<0.05);and compared with the OVX,in the 3 medication groups,BMSCs PPARγ m RNA`s level decreased significantly(P<0.01);and compared with the BJL group,the expression of PPARγ m RNA in Yougui pillincreased significantly(P<0.01);and compared with Zuogui pill group,the expression of BMSCs PPARγ m RNA in Yougui pill group increased significantly(P<0.01).Protein test results showed that,compared with the control group,in the OVX and Zuogui pill,Yougui pill group,the expression levels of BMSCs PPARγ protein all had significant difference(P<0.05);and compared with the model group,in the medication group,the expression levels of BMSCs PPARγ protein were all significantly lowered(P<0.05);and compared with the BJL group in Yougui pill group PPARγ protein was significantly increased(P<0.05);and compared with Zuogui pill group,the expression of BMSCs PPARγprotein in Yougui pill group increased significantly(P<0.01).3.3.5 Zuogui pill,Yougui pill on the effect to the expression m RNA and protein of ovariectomized rats BMSCs LPL m RNA test results show that,compared with the control group,in the OVX,the expression of BMSCs LPL m RNA was significantly increased(P<0.01);LPL m RNA was significantly decreased in Yougui pill group(P<0.05);and compared with the OVX,in the medication group the expression of BMSCs LPL m RNA were significantly lowered(P<0.01);and compared with the BJL group,the expression of LPL m RNA in Yougui pill group was significantly decreased(P<0.05);and compared with Zuogui pill group,in Yougui pill group the expression of BMSCs LPL m RNA was significantly decreased(P<0.05).Protein test results show that,compared with the control group,in the model group and Yougui pill group,the expression level of BMSCs LPL protein had significant difference(P<0.05);and compared with the model group,in the medicaton group,the expression level of BMSCs LPL protein all decreased significantly(P<0.01);and compared with the BJL group,in Yougui pill group significantly decreased the expression of LPL protein(P<0.01);and compared with Zuogui pill group,the expression of BMSCs LPL protein significantly decreased in Yougui pill group(P<0.01).4 Effect on Verbascosideto the apoptosis of h FOB1.19 cells4.1 The effect on Verbascosideto the cell proliferation of h FOB1.19 The growth curve of h FOB1.19 was S shape,and with the inoculative concentrationincreased,the optical density rose with it.The incubation period was 3h after the cell inoculation,and after 12 h,the cells began to enter into log phase,the cell growth reached peak at 48 h.The results show that,compared with the control group,with the increase of Verbascoside concentration,the cell proliferative ability of h FOB1.19 reduced 10-6M and10-5M of Verbascoside.Verbascoside increased the proliferative activity of h FOB1.19,and the 10-5M of verbascoside.Verbascoside had the effect of promoting cell proliferation largelist.4.2 The optimal conditions for h FOB1.19 cell apoptosis induced by dexamethasone The results of Annexin V/PI double staining showed that,through flow diagram,comparingthe flow rate of the right lower quadrant and right upper quadrantit was the most suitable condition that the concentration of dexamethasone was 10-5 mol·L-1for 48 h.4.3 The effects on Verbascoside to the cell apoptosis of h FOB1.194.3.1 The effect on different concentration of acteoside Verbascoside to the cell apoptosis of h FOB1.19.The results of Annexin V/PI double staining showed that,through flow diagram,the comparing the flow rate of the right lower quadrant and right upper quadrant and compared with the control group,the apoptosis of 2*10-4M and 3*10-4M had significant difference(P<0.05)with the Verbascoside.4.3.2 The effect on different concentration of Verbascosideto the cell apoptosis of h FOB1.19 by Hoechst33258The results showed that,in the control group,the cells chromatin was evenly distributed,showing a blue fluorescence diffusely low density.The Verbascoside in different concentration streated h FOB1.19 cells with varying degrees of the characteristics of apoptosis appearing.Verbascoside of 2*10-4M and 3*10-4M obviously promoted the apoptosis.Especially the Verbascoside granular fluorescence of 3*10-4M was significantly higher than the other groups.With the increase of Verbascoside concentration,the apoptosis also increased gradually.4.3.3 The effect of different concentration of Verbascoside on the apoptosis of TUENEL h FOB1.19 cells The results showed that,compared with the control group,with the increase of Verbascoside concentration,the number of green gradually increased fluorescence.the Verbascoside of 2*10-4M and 3*10-4M could obviously promote the apoptosis.Especially Verbascoside of 3*10-4M,the number of green coloring cells more significantly increased than the other groups.With the increase of Verbascoside concentration,the apoptosis increased gradually.4.4 The study on the mechanism that the Verbascoside intervenedthe apoptosis cells of h FOB1.194.4.1 The effect of the Verbascoside to h FOB1.19 cell Bcl-2/ Bax m RNA and protein m RNA test results showed that,compared with the control group,the Verbascoside concentration of 2*10-4M,3*10-4M and Bcl-2/Bax,h FOB1.19 cells of Dex m RNA the expressions were significantly different(P<0.05);compared with Dex,Verbascoside of 10-6M,10-5M,Bcl-2/Bax 10-4M of h FOB1.19 cells of Bcl-2/Bax m RNA,the expressions were significantly different(P<0.05);and compared with 3*10-4M,Verbascoside of 10-6M,10-5M,10-4M Bcl-2/Bax m RNA,the expression had significant difference(P<0.05).Protein detection results show that the ratio of Bcl-2/Bax decreased,compared with the control group,Verbascoside concentration 10-4M,2*10-4M,Bcl-2/Bax 3*10-4M expression of h FOB1.19 cells and Dex protein were significantly different(P<0.05);compared with Dex,Verbascoside 10-6M,10-5M,Bcl-2/Bax expression in h FOB1.19 cells 10-4M protein are significant differences(P<0.05);compared with 3*10-4M,Verbascoside 10-6M,10-5M,10-4M,Bcl-2/Bax protein expression had significant difference(P<0.05).4.4.2 The expression of h FOB1.19 cell Casepase-3 protein Verbascoside The results show that,compared with the control group,2*10-4M,3*10-4M and Dex expression of h FOB1.19 cells Casepase-3 protein significantly increased(P<0.05);compared with Dex,10-6M,10-5M and 10-4M expression of h FOB1.19 cells Casepase-3 protein significantly decreased(P<0.05);compared with 3*10-4M,Verbascoside 10-6M,10-5M,10-4Mexpression of h FOB1.19 cells Caspase-3 protein decreased significantly(P<0.05).4.5 Effect of Verbascoside on ERK1 / 2 Signaling Pathway in h FOB1.19 Cells4.5.1 Effect of Verbascoside on ERK1 / 2 Protein Phosphorylation in h FOB1.19 Cells The results showed that 3 * 10-4M and Dex significantly enhanced the expression of p-ERK protein in h FOB1.19 cells compared with the group group(P<0.05).Compared with Dex group,h FOB1 was found in 3 * 10-4M group(P<0.05).Compared with 3 * 10-4M,the expression of p-ERK protein in h FOB1.19 cells was decreased(P<0.05),and the expression of p-ERK protein was significantly decreased(P<0.05).In addition,the expression of p-ERK was significantly down-regulated in PD98059 group(P<0.05).4.5.2 Effects of Verbascoside on ERK1 / 2 signaling pathway Bcl-2 protein in h FOB1.19 cells The results showed that the expression of Bcl-2 protein in h FOB1.19 cells was significantly up-regulated by Control + PD,2 * 10-4M,3 * 10-4M,3 * 10-4M + PD and Dex groups compared with blank group(P <0.05).Compared with Dex,the expression of Bcl-2protein in h FOB1.19 cells was down-regulated by 3 * 10-4M,3 * 10-4M,3 * 10-4M + PD and Dex groups,(P<0.05).In addition,after adding PD98059,the expression of Bcl-2 protein was significantly higher in each group than that without PD98059,and the expression of Bcl-2protein in h FOB1.19 cells was significantly higher than that in PD98059 group protein expression was significantly down-regulated(P<0.05).Conclusion:1 Zuogui and Yougui pill can promote the proliferation of BMSCs in ovariectomized rats,and Zuogui pill promote the proliferation of BMSCs in ovariectomized rats.2 The mechanism of postmenopausal osteoporosis is associated with osteogenesis and adipogenesis imbalance.Zuogui and Yougui pill can regulate the expression of BMSCs into osteoblasts and adipogenic differentiation,and the Zuogui and Yougui pill can promote the expression of osteogenesis-related indexes.Yougui pill can promote the expression of adipogenic related indexes,and Zuogui pill inhibit adipogenic differentiation.3 Zuogui and Yougui pill can inhibit the ovariectomy BMSCs osteogenesis,post-adipogenic differentiation of apoptosis,and Zuogui pill beneficial to osteogenic differentiation,Yougui pill beneficial to adipogenic differentiation.4 With the increase of Verbascoside concentration,the apoptotic condition increased with the increase of the concentration,when Verbascoside concentration is 3*10-4mol/L the strongest apoptosis.5 Effect of ERK1 / 2 Signaling Pathways on the apoptosis of h FOB1.19 Cells. |
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