Based On The Expression Of TRAF6 And TAK1, The Effect Of Jingang Wan On PMOP Rats With Kidney-yang Deficiency Was Investigated

Objective:To study the expression of TRAF6 and TAK1 in rankl-rank-opg pathway,and to explore the mechanism of Jingang pill in the prevention and treatment of kidney yang deficiency syndrome in postmenopausal osteoporosis(PMOP).Methods:Seventy SPF grade 6-month-old female SD rats were randomly divided into blank control group(NTC n=10),sham operation group(sham n=10)and ovariectomized group(OVX n=50).OVX group used bilateral ovariectomy to establish postmenopausal osteoporosis rat model,sham group removed abdominal adipose tissue with the same weight of ovaries,OVX group was given hydrocortisone injection intramuscularly one week after operation to maintain kidney yang deficiency model.The OVX group was randomly divided into model group,estradiol valerate group and Jingang pill high,medium and low dose groups,with 10 rats in each group.Each group was given corresponding dose and concentration of drugs by gavage once a day for 12 weeks The model group was also given the same volume of distilled water.After 6 and 12 weeks of administration,the expression of rank,RANKL,OPG,TRAF6 and TAK1 in serum of rats in each group was detected by ELISA,and the relative expression of key signal RANKL,OPG,TRAF6 and TAK1 mRNA was detected by PCR products;after 12 weeks of administration,the bone morphology of PMOP model rats with kidney yang deficiency was observed by HE staining.Results:(1)BMD and serum ELISA detection: After 8 weeks of modeling,compared with the blank control group and the sham operation group,the BMD of the model group was significantly decreased(P<0.05).At 6 and 12 weeks after administration,there was no significant difference in serum OPG,rank,RANKL,TRAF6 and TAK1 between the blank group and the sham operation group(P>0.05);compared with the model group,the serum OPG content of the other groups increased,while the content of rank,RANKL,TRAF6 and TAK1 decreased,with significant difference(P<0.05).Compared with the high-dose group,the serum OPG content in the high-dose group increased,while the levels of rank,RANKL,TRAF6 and TAK1 decreased,with statistical significance(P<0.05);the OPG content in the estradiol valerate group was lower than that in the high-dose group,and the levels of rank,RANKL,TRAF6 and TAK1 were higher than those in the high-dose group,while there was no statistical difference between the estradiol valerate group and the medium and low-dose groups The difference was statistically significant(P>0.05).(2)PCR detection and he staining detection of femur tissue: at 6 and 12 weeks after administration,there was no significant difference in OPG,RANKL,TRAF6 and TAK1 between the blank group and the sham operation group(P>0.05);compared with the model group,the mRNA content of OPG in the other groups increased,while the relative expression of RANKL,TRAF6 and TAK1 decreased,with significant difference(P<0.05)Compared with the estradiol valerate group,the indexes of high-dose Jingang pill group were improved more significantly(P<0.05);compared with the middle and low-dose Jingang pill groups,the content of OPG in femur of high-dose group was increased,and the content of RANKL,TRAF6 and TAK1 was decreased,with statistical significance(P<0.05);there was no significant difference between the estradiol valerate group and the middle and low-dose Jingang pill groups Statistical significance(P>0.05).The results of he pathological section suggest that Jingang pill can improve the morphology and structure of trabecular bone and adjust the arrangement of cells in bone tissue.Conclusion:The intervention of Jinggang pill can reduce the mRNA expression of the key factors TRAF6 and TAK1,up regulate the OPG content in serum and femur tissue and decrease the expression of rank and RANKL,thus play a role in preventing and treating osteoporosis of kidney yang deficiency type postmenopausal osteoporosis.The results suggest that the decrease of expression of TRAF6 and TAK1 may be one of the mechanisms of Jingang pill in preventing and treating the deficiency of kidney yang in postmenopausal osteoporosis.The increase of dosage and course of Jinggang pill can promote the decrease of expression of TRAF6 and TAK1.