Study On The Influence Of Dampness-heat Syndrome To Bone Metabolism And Hypothalamic,Pituitary,Target Gland Morphology In Postmenopausal Rats

Objective:By testing bonemineral density and bone metabolism related indicators,observed the changes of morphology of Hypothalamic,Pituitary,Target Gland changes of Ovariectomized rats,preliminary study the role of dampness heat syndrome in the development of osteoporosis in postmenopausal rats and explored the possible mechanism in the occurrence and development of postmenopausal osteoporosis.Methods:（1）30 SD rats were randomly divided into Normal control group,Dampness heat syndrome model group and the Syndrome of drug intervention group,with 10 rats in each group.Normal control group placed in room temperature,common feed,the Dampness heat syndrome model group was placed in damp heat environment and fed with damp heat diet for 24days;Syndrome of drug intervention group was given Dampness heat syndrome modeling factors while intervention with Dampness heat syndrome adjustment prescription.Appearance of rats observed and photo recorded,quantitative analysis with Photoshop Image Processing Software,measured body weight,food intake and calculate food potency,scanning electron microscope observation of tongue surface,testing TC,TG,HDL-c,LDL-c of serum by microplate method.（2）100 SD rats were randomly divided into Normal control group,Sham operation group,OVX model group and OVX+Dampness heat syndrome model group,with 20 rats in each group.Bilateral ovaries were removed in OVX model group and OVX+Dampness heat syndrome model group（dampness heat+damp heat diet）,the other three groups were fed with normal diet at room temperature.Observation of appearance,by Micro-CT measured Bone mineral density,Bone microstructure parameter;by microplate method testing of calcium and phosphorus in blood.Serum E2,1,25（OH）₂D₃,PTH,ACTH and CORT were detected by ELISA.By HE Coloration and Transmission Electronic Microscopy technique was observed the morphological structure of Hypothalamus,Pituitary,target gland.Results:（1）Compared with Normal control group,Dampness heat syndrome model group showed fast-response,restlessness,rough hair,ear nose claw color deepened,thick tongue,scanning electron microscopy showed that the filiform papillae were obviously"broken",number was increased（P<0.05）,the fungiform papilla stratum corneum thickened and taste hole disappeared.urine colors was deepen,defecation dry and hard,weight gain,diet water reduced（P<0.05）,Serum TC,TG levels increased.Appearance characterization of Syndrome of drug intervention group close to Normal control group and there are some differences compared with Dampness heat syndrome model group,tongue filamentous papillae number was reduced,fungal papilla cuticle was thinned.（2）Compared with Normal control group,Sham operation group and OVX model group after operation 7w,OVX+dampness heat syndrome group BMD,BV,Tb.N,Tb.Th decreased,Serum Ca²⁺and PTH increased,E2 and CORT decreased after operation 7w and 9w（P<0.05）.（3）Light microscope showed that,compared with OVX model group,OVX+dampness heat syndrome group hypothalamic astrocyte swelling,nuclear enlargement,chromatin darker,synaptic thickening,vascular interstitial hyperemia,clear boundary of pituitary cytoplasm,increased number of basophil granulocytei and blood sinus,hyperemia,inflammatory cell infiltration.Adrenocortical Hyperplasia,cells arrangement irregular,cortical cell cytoplasm rich in lipid droplets.（4）Transmission electron microscopy showed that,compared with OVX model group,OVX+dampness heat syndrome group increased heterochromatin and autolysosomes in hypothalamic neurons,rough endoplasmic reticulum expansion and degranulation,postsynaptic membrane mitochondria swelling,increased number of lysosomes and lipofuscin.Increased number of pituitary mitochondria,swelling,breakage.Heterochromatin accumulation in nucleus of adrenal cortex,increased number of mitochondria,rough endoplasmic reticulum expansion,secretion of large particles.Conclusions:（1）According to the clinical causes of dampness heat syndrome,dampness heat syndrome animal models can be established.The animal model showed similar biological characteristics of clinical dampness heat syndrome and had stability and repeatability.（2）The causes of dampness heat syndrome may accelerate the reduction of bone mineral density,damage bone microstructure and shorten the onset time of osteoporosis by affecting the levels of serum CORT,PTH,Er and blood calcium in postmenopausal rats.（3）Causes of dampness heat syndrome may change the pathological and ultrastructural of hypothalamus and pituitary gland in postmenopausal rats.These changes caused by dampness heat syndrome may be one of the biological basis of dampness heat syndrome affecting the body and bone tissue of postmenopausal rats.