Effects Of Pain-relieving Capsule On Regulating MAPK Pathway And ERK1/2, P38MAPK And Its Phosphorylated Proteins In Ovariectomized Rat

Objective: To research the impact of Zhuanggu Zhitong Capsule on the expression of ERK1/2,p38 MAPK and p-ERK1/2,P-P38 MAPK in ovariectomized female rats,and to explore the mechanism of Zhuanggu Zhitong Capsule regulating MAPK pathway in the intervention of postmenopausal osteoporosis.Methods:60 female250 g SD rats were randomly divided into 6 groups: Blank,Shamoperation,Model,Zhuanggu Zhitong Capsule,RAPA and 3-MA group.The blank group did’t treat,and the Sham removed a little fat around the ovary.The other 40 ovaries were removed to establish the ovariectomy model.The corresponding drugs were given 7 days after operation for 13 weeks.The samples were taken after the last intragastric administration.Thepathological morphology of bone tissue was observed by HE staining;the content of serum E2 was detected by ELISA;the expression of ERK1/2,p-ERK1/2,p38 MAPK and p-p38 MAPK protein in femur was analyzed byimmunohistochemistry.Results:(1)The rats in the normal and the sham-operation group had stable estrous cycle,while the rats in the model group the rut was disordered,it showed successful.(2)HE: It’s damaged of glomerular structure in model and 3-MA group the epithelial cells of renal tubules were swollen and basement membrane thickened,some renal tubules were obviously atrophic,and lots of inflammatory cells were found in renal stroma.The mesangial stroma and cell proliferation were increased;the glomerulus and tubules in the blank and sham-operation group were intact,and the mesangial cells were arranged in order without abnormal changes;the zhuangguozhitong capsule and RAPA group could.The glomeruli were intact,and the renal tubules were’t swollen,no tubular atrophy and only a spot of inflammatory cells in it(3)Bone HE:in sham-operation,blank and RAPA groups,the bone marrow cavity of Zhuanggu Zhitong Capsule group had no change,the distribution density of bone trabeculae was uniform,and the periosteum was intact.In the model and 3-MA group,the medullary space was larger,and there was umcomplete periosteum..(4)ELISA: The E2 in the Sham-operation was’ t statistically significant compared to the blank group(P > 0.05).The E2 in the model group was lower compared to the Sham-operation(P < 0.01).The E2 of Zhuanggu Zhitong Capsule and RAPA group were higher compared to the model(P < 0.01).Compared to 3-MA,E2 in Zhuanggu Zhitong Capsule and RAPA group was higher(P < 0.01)(5)Immunohistochemistry :there’s no difference in the each protein between the blank and the sham-operation group(P > 0.05).Compared to the sham-operation,model ’s each protein expression in kidney and bone tissue was lower(P < 0.01).Compared to the model group,the expression of protein in Zhuanggu Zhitong Capsule and RAPA group increased(P <0.01);the gray value expression of protein in kidney and bone tissue in 3-MA reduced compared to the model(P < 0.01);compared to the 3-MA,the gray value expression of protein in Zhuanggu Zhitong Capsule group and RAPA group increased,with difference(P < 0.01)There was no significant difference in the expression of ERK1/2,p38 MAPK and phosphorylated protein between the two groups(P > 0.05).Conclusion:(1)Zhuanggu Zhitong Capsule can regulate the MAPK pathway of ovariectomized rats,positively regulate the expression of ERK1/2,p38 MAPK and their phosphorylated proteins(p-ERK1/2,p-p38MAPK)and intervene postmenopausal osteoporosis;(2)Zhuanggu Zhitong Capsule has the same effect as estradiol in promoting bone formation;(3)Zhuanggu Zhitong Capsule can effectively improve the bone microstructure of PMOP.