| Research objectives:To study the effect of Mongolian medicine Runguling on the treatment of postmenopausal osteoporosis,use different treatment methods of Mongolian medicine,observe the general situation of female C57BL/6 mice ovariectopic osteoporosis model,bone density,and the content and expression of specific proteins related to bone metabolism,explore the influence of Mongolian medicine Runguling on bone metabolism,and clarify the mechanism of action in the treatment of postmenopausal osteoporosis.It provides certain theoretical and scientific application value for Mongolian medicine Runguling in the treatment and prevention of postmenopausal osteoporosis.Research methods:The 6-month-old SPF female C57BL/6 mice were selected to establish the postmenopausal osteoporosis model by ovarectomy.The mice were randomly divided into 7groups(10 in each group): normal group,sham operation group,model group,Runguling high-dose group,Runguling medium-dose group,Runguling low-dose group,and alendronate sodium tablet group.The other groups except the normal group and the sham group underwent bilateral oophorectomy to establish the postmenopausal osteoporosis model.Normal group was given normal maintenance feed,sham operation group,model group was given normal saline intragastric administration,Runguling high-dose group,Runguling medium-dose group,Runguling low-dose group,and alendronate sodium tablet group were given corresponding drug treatment,starting from the 7th day after operation and being administered on time at 9:00 AM daily for 12 weeks.After 12 weeks,the drug was stopped,and the serum,kidney and femur of mice were collected for use after 24 hours of fasting and water prohibition.Dual energy X-ray bone mineral density detector was used to detect the bone mineral density of mouse right femur in vitro,ELISA was used to detect the protein content of PINP,BALP,β-CTX,TRACP,etc.,to verify the establishment of osteoporosis model and the effect of medication in each group.The expression of Wnt3 a m RNA andβ-catenin in the right kidney and left femur tissues of mice in each group was detected by ELISA,and the expression of Wnt3 a mrna and β-catenin in the right kidney and left femur tissues of mice in each group was detected by RT-PCR.All data were statistically analyzed by SPSS26.0 statistical software.To observe the effect of Runguling on the expression of bone metabolism related proteins in kidney and bone tissue,and analyze the mechanism of Runguling on postmenopausal osteoporosis.Research results:1.General condition,body weight change,uterine index No mice died during the whole postoperative and gavage experiment.The general situation of each group of mice was observed carefully one week after the operation.The normal group of mice were active,alert,active,responsive,smooth hair,normal diet and sleep.Compared with the normal group,the spirits of mice in the sham operation group and other groups were lower,their reactions were slower,there was no hair in the operation area,and the hair in the rest areas was bright and smooth.One week after surgery,the incision healed completely and the hair grew out.It could not be identified whether the operation was performed or not,and the condition basically returned to normal.During the whole process of gavage treatment,the mice in each group had stable mood,no stress reaction was observed,and their mental state was better and more active,with more agile reaction and smooth hair.After 12 weeks of intragastric administration,the body weight of mice in the normal group increased slightly with the feeding time,which was in line with the general law of animal natural growth.Compared with normal group,the body weight of mice in all groups decreased significantly after surgery(P < 0.05).The body weight of the sham operation group was gradually close to that of the normal operation group(P > 0.05).The body weight of the model group was compared two months after the operation.The body weight of the other drug administration groups was slightly higher(P < 0.05),but lower than that of the normal and sham operation groups(P < 0.05).After 12 weeks of intragastric administration,the V-shaped uterus was completely removed from the lower abdomen,and the surrounding adipose tissue was carefully removed to ensure the integrity of the uterus.The weight record was measured by the Tenson’s scales.The results were as follows: there was no significant difference between the normal group and the sham group,and there was significant difference between the model group and the other administration groups(P < 0.05).2.The bone mineral density of the right hind limb of mice in each group was measured by dual-energy X-ray bone mineral density tester.The results showed that there was no significant difference in BMD value between the sham operation group and the normal group(P > 0.05).Compared with normal group,BMD value in all administration groups was decreased(P < 0.05).Model group had the lowest value,and compared with model group,all groups were significantly increased(P < 0.05).Compared with Runguling medium dose group,Runguling high dose group and Runguling low dose group had significant differences(P <0.05);Compared with alendronate sodium group,Runguling medium dose group had significant difference(P < 0.05);3.Compared with the normal group,the serum PINP and BALP contents in the sham group had no significant changes(P > 0.05);Compared with normal group,the values of PINP and BALP in administration groups were significantly decreased(P < 0.05);Compared with model group,PINP and BALP of all groups were significantly different(P < 0.05).Compared with Runguling medium dose group,Runguling high dose group and Runguling low dose group had significant differences in PINP content(P < 0.05);Compared with Runguling medium-dose group,Runguling high-dose group and Runguling low-dose group had no significant difference in BALP content(P > 0.05);Compared with alendronate sodium group,PINP and BALP in Runguling medium dose group were significantly different(P <0.05);4.Compared with normal group,the serum β-CTX and TRACP contents in sham group had no difference(P > 0.05);Compared with normal group,β-CTX and TRACP values in administration groups were significantly decreased(P < 0.05);Compared with model group,β-CTX and TRACP were significantly different in all groups(P < 0.05).Compared with Runguling medium-dose group,there were significant differences in β-CTX and TRACP in Runguling high-dose group(P < 0.05),but no significant differences in Runguling low-dose group(P > 0.05);TRACP of Runguling low-dose group was significantly different(P < 0.05).Compared with alendronate sodium group,there were significant differences in β-CTX and TRACP in Runguling medium dose group(P < 0.05);5.There was no significant difference in Wnt3 a protein content in bone and kidney of mice in sham operation group compared with normal group(P > 0.05);Compared with normal group,there were significant differences among administration groups(P < 0.05).Compared with model group,all groups had significant differences(P < 0.05).There was no significant difference between Runguling high-dose group and Runguling medium-dose group(P > 0.05),but there was significant difference between Runguling low-dose group and Runguling medium-dose group(P < 0.05).Compared with alendronate sodium group,Runguling medium dose group had significant difference(P < 0.05);6.The expression of Wnt3 a m RNA and β-cateni protein in bone tissue of mice in the experimental group had no significant differences compared with the normal group(P > 0.05);Compared with normal group,there were significant differences among administration groups(P < 0.05).Compared with model group,all groups had significant differences(P < 0.05).The m RNA expression of Wnt3 a in Runguling medium dose group was significantly different from that in Runguling high dose group(P < 0.05);,Runguling low-dose groups had no significant difference(P > 0.05);The expression of β-catenin in Runguling medium dose group was significantly different from Runguling high dose group and Runguling low dose group(P < 0.05);Compared with alendronate sodium group,Runguling medium dose group had significant difference(P < 0.05);7.The expression of Wnt3 a m RNA and β-cateni protein in kidney tissue of mice in each group had no significant difference compared with that in normal group(P > 0.05);Compared with normal group,there were significant differences among administration groups(P < 0.05).Compared with model group,all groups had significant differences(P < 0.05).Compared with Runguling medium dose group,Runguling high dose group and Runguling low dose group had significant differences(P < 0.05);Compared with alendronate sodium group,Runguling medium dose group had significant difference(P < 0.05);Research conclusion:1.Mongolian medicine Rungu Ling can improve the bone mineral density and serum type Ⅰ collagen amino terminal prolongating peptide(PINP),bone alkaline phosphatase(BALP)protein content,reduce the β-collagen special sequence(β-CTX),tartrate-resistant acid phosphatase(TRACP)protein content of postmenopausal osteoporosis mice,and the high dose of Mongolian medicine Rungu Ling has the best effect.Medium dose followed,low dose had the least effect.It is proved that Mongolian medicine Runguling can improve bone density,affect bone formation and bone destruction protein in serum,so as to achieve the purpose of treating postmenopausal osteoporosis.2.Mongolian medicine Runguling can improve the specific Wnt3 a protein content in bone tissue and kidney tissue of postmenopausal osteoporosis mice by activating the Wnt/β-catenin classical pathway,and can improve the expression of Wnt3 a m RNA andβ-cateni.It was proved that Mongolian medicine Runguling can improve bone cell metabolism and play a role in the prevention and treatment of postmenopausal osteoporosis. |
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