Study On The Influence Of Dampness-cold Syndrome On Bone Mineral Density And Hypothalamus,pituitary,target Gland Morphology In Postmenopausal Osteoporosis Rats

Objective:Discussing the effects of dampness-cold syndrome for traditional Uyghur medicine on the development of osteoporosis in ovariectomized rats by detecting the bone density and bone metabolism,and observing the morphology and structure of hypothalamus,adenohypophysis and target glands.Methods:（1）30 female SD rats were randomly divided into normal control group,dampness-cold syndrome group and drug intervention group.Normal control group was given ordinary food and appropriate environment,and the dampness-cold syndrome group was given dampness-cold feeding environment with foods and cold water swimming for 24 days.The drug intervention group adopted the above modeling factors while giving the drug which prevent dampness-cold syndrome;The appearance sign were observed and quantified,body weight,food intake,water intake,nutritive value of food were measured,morphological changes of the tongue were scanning by electron microscope.（2）80 female SD rats were randomly divided into normal control group,sham operation group,OVX model group,and OVX+dampness-cold syndrome group,with 20 rats in each group.Except OVX+dampness-cold syndrome group,the other three groups were given ordinary food and appropriate environment.The bilateral ovaries of OVX model group and OVX+dampness-cold syndrome group were removed,and the OVX+dampness-cold syndrome group was given dampness-cold feeding environment with foods;Bone mineral density and bone microstructure parameters were determined by micro-CT.The levels of ACTH,CORT,Ca,P,1,25-（OH）₂D₃,E₂and PTH in serum were detected by ELISA.The morphological changes of hypothalamic-pituitary-target gland（adrenal gland and thyroid gland）were observed by HE staining and transmission electron microscopy.Results:（1）Building of model 24 days,dampness-cold syndrome group showed the reaction is slow,tired,hair messy and lackluster,lethargy,appeared whitish coating of the tongue,slight white nose and claws,heavy urine volume and wet stools,general performance score reached 80.10.Compared with normal control group,weight growth speed was slow,food intake increased,water intake decreased（P<0.05）,the tongue tissue was keratinized;Compared with dampness-cold syndrome group,drug intervention group physical appearances was close to control group,the degree of tongue keratinization was lower.（2）Compared with OVX model group,the activity of rats in OVX+dampness-cold syndrome group was significantly reduced,showed tiredness,whitish coating of the tongue were more obvious,stools were not formed.（3）Compared with normal control group,sham operation group and OVX model group,bone mineral density（BMD）,trabecular thickness（Tb.Th）,trabecular number（Tb.N）were decreased and trabecular septum（Tb.Sp）of rats in OVX+dampness-cold syndrome group were widened at 7 and 9 weeks after operation（P<0.05）;Compared with OVX model group,the BMD,Tb.Th,Tb.N of OVX+dampness-cold syndrome group decreased and Tb.Sp widened at week 7（P<0.05）;Compared with 7th week,there was significant difference in the BMD,Tb.Th,Tb.N,Tb.Sp of OVX+dampness-cold syndrome rats at the 9th week（P<0.05）.（4）On the 7th and 9th week after surgery,compared with OVX model group,the concentration of CORT and 1,25-（OH）₂D₃in OVX+dampness-cold syndrome group were increased（P<0.05）.（5）Compared with OVX model group,the light microscopy results showed that the hypothalamic interstitium of rats in OVX+dampness-cold syndrome group showed edema,pituitary interstitium showed vacuolation,adrenal medulla lipid droplet hyperplasia,and thyroid follicular cell infiltration.The results of electron microscopy showed that compared with OVX model group,mitochondria of hypothalamic neurons,pituitary ACTH cells,adrenal cortical cells and thyroid follicles were swollen in OVX+dampness-cold syndrome group.Conclusion:（1）Starting with the main causes of clinical dampness-cold syndrome,it is feasible to simulate the animal model of dampness-cold syndrome.The model is reliable and stable.（2）Dampness-cold syndrome affected the levels of CORT and 1,25-（OH）₂D₃in ovariectomy-induced osteoporosis rats,accelerated the decrease of bone mineral density,damaged bone microstructure,and advanced the onset time of osteoporosis.（3）Dampness-cold syndrome caused changes in hypothalamus,adenohypophysis and target gland histology and cell ultrastructure in ovariectomy-induced osteoporosis rats.The change may be one of the biological basis of the influence of the etiological factors of dampness-cold syndrome on the body and bone tissue,leading to dampness-cold temperament disorder and functional disorder.