| Postmenopausal osteoporosis is common and frequent disease in older women. Gu kang has good effect on prevention and treatment of osteoporosis. The specific mechanism of it has not yet been explained clearly, especially the effect on adipogenic differentiation of MSCs.ObjectiveCulturing the rats'MSCs of Osteoporosis in vitro. Promoting differentiation of MSCs into adipose cell with adipogenic differentiation inducer. Investigating the effect of Gukang drug-containing serum on the expression of PPARγ'genes during the adipogenic differentiation of MSCs, with different concentrations of Gukang drug-containing serum intervention, and nylestriol drug-containing serum intervention and comparison.MethodsAnimal studies:24 female SD rats of SPF rank, which were underwent bilateral ovariectomy and regular feeded for 12 weeks.3 were selected randomly to prepare nilestriol drug-containing serum.9 were fed with different doses of Gukang to prepare drug-containing serum.12 were killed to prepare MSCs. MSCs were randomly divided into 8 groups after purification, namely:model group (MG), adipogenic induction group(AIG), western medicine group(WMG), adipogenic western medicine group (AWMG), herb group (HG), adipogenic low dose herb group(ALDHG), adipogenic medium dose herb group(AMDHG), adipogenic high dose herb group(AHDHG). Each group make oil red 0 stain which were observed under light microscopy in adipogenic 7,14 days. The mRNA expression of PPARγwas detected by RT-PCR, the PPARγprotein content was detected by western blot. Clinical Research:Since the beginning of March 2010 to October 2010, outpatient and in-patient departments our Hospital, confirmed by clinical examination in postmenopausal patients with osteoporosis 87 patients were choiced.10 h ealthy 20-30 year-old unmarried young women as control group (A group), the serum levels of PPARγof eligible subjects and healthy women were tes ted. Every 5 years as a grade, Group of patients with osteoporosis are gr ouped according to menopause duration, divided into 6 groups, respectivel y, B, C, D, E, F group, which corresponds to the order of 2-5 years of me nopause,5-10,10-15 years,15-20 years, more than 20 years. The PPARγin serum levels of subjects and menopause duration were observed and the co-rrelation between the two were Explored.ResultsAnimal studies(A) AdipogenesisInterventions applied in each group after 7 days, compared with MG, AIG showed significant difference (P<0.01), AWMG, adipogenic herb groups showed significant difference (P<0.05). Compared with AIG, AWMG, ALDHG and AMDHG showed significant difference (P<0.05), AHDHG showed high significant difference (P<0.01). there are no difference between WMG and HG (P> 0.05). nor AWMG or adipogenic herb groups(P>0.05).Intervention for 14 days, compared with MG, AIG showed significant difference (P<0.05), other groups showed no significant difference (P>0.05). Compared with AIG, the adipogenesis rate of WMG and HG was significantly lower than AIG(P<0.01). AWMG, adipogenic herb groups had significant difference compared with AIG(P<0.05). The adipogenesis rate of HG was significantly lower than that of ALCHG, AMCHG and AHDHG (P<0.05). The comparison between 7 days and 14 days after intervention showed that adipogenesis rate of each group after 14 days increased than that of 7 days, but there was no statistically difference(P> 0.05).(B) PPARγmRNA expressionAfter 7 days'intervention, the PPARγmRNA expression of WMG, AWMG and AHDHG were decreased than that of MG (P<0.05), ALDHG was significantly lower (P<0.01), the PPARγmRNA expression of HG and AMDHG also tended to decrease, but there was no significant difference compared with MG(P>0.05). The PPARγmRNA expression of ALDHG was significantly lower than that of AIG(P<0.01), which was higher than that of AHDHG and AWMG (P<0.05). There was no statistically significant in remaining groups (P>0.05). WMG was better than HG on suppressing the expression of PPARγmRNA (P<0.05). There was no significant difference between AWMG and each adipogenic herb group(P>0.05). Compared with HG, There was highly significant or significant differences on suppressing the expression of PPARγmRNA in ALDHG,AHDHG and AWMG (P<0.01, P<0.05). Among adipogenic herb groups, effection of suppressing the expression of PPARγmRNA in ALDHG was the most obviously (P<0.01, P<0.05), followed by AHDHG (P<0.05).After 14 days'intervention, in addition to AIG, the PPARγmRNA expression of the other groups compared with MG showed significant differences (P<0.05). Compared with AIG, WMG, AWMG and AMDHG showed a significant difference (P<0.05), ALDHG and AHDHG were analyzed with the very significant difference (P<0.01). There was no significant expression differences in expression of PPARγmRNA between WMG and HG (P>0.05). The PPARγmRNA expression of ALDHG and AHDHG was significantly lower than that of AWMG (P<0.01). Compared with HG, ALDHG and AHDHG had significant difference (P<0.05). Among adipogenic herbs groups, effection of suppressing the expression of PPARγmRNA in ALDHG was the most obviously (P<0.01), followed by AHDHG (P<0.05).After 7 days'and 14 days'intervention, the PPARγmRNA expression of AIG was found significantly increased (P<0.05). The adipogenesis by oil red staining after 14 days'intervention also increased, but the statistics analysis was not significant (P>0.05). The PPARγmRNA expression of other groups also decreased, but showed no significant difference (P>0.05).(C) PPARγprotein detectionAfter 7 days'intervention, the expression of PPARγprotein in ALDHG and AHDHG was the lowest, followed by AWMG. That in HG and AMDHG was the highest, followed by AIG. After 14 days'intervention, the expression of PPARγprotein in MG and HG was the highest, followed by AIG. That in ALDHG and AHDHG was the lowest.After 7,14 days'intervention, expression of PPARγprotein in each group showed that MG and AIG were significantly higher than those. The expression of PPARγprotein decreased after 7,14 days'intervention in HG, which was still higher than that in other groups. The expression of PPAR Y protein in ALDHG and AHDHG was the lest, followed by AWMG.Clinical ResearchThe PPARγinserum levels in postmenopausal women has significantly i-ncreased in the first 15 years, and fifteen years after the menopause ha s stabled (Comparison between B, C, D and A, E, F, P<0.05; among the three groups, B, C, D, P>0.05; among the three groups, A, E, F, P> 0.0 5;). By correlation analysis, serum PPAR Y concentration and menopause dur-ation has not been significantly correlation, Spearman correlation coef ficient yp= 5.870, After testing correlation coefficient P> 0.05.ConclusionAnimal studiesLow and high drug-containing serum of Gu kang can reduce the number of fat cells generated from MSCs, reduce the expression of PPARγmRNA and its protein. They can prevent adipogenic differentiation of osteoporosis rat MSCs, promote their osteogenic differentiation. The efficacy is similar with that of nilestriol.Clinical Research:The first fifteen years in postmenopausal serum PPARγlevels were si gnificantly higher than healthy women. Fifteen years after the serum cone entrations of PPARγof the menopause stablilized.
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