| Objective: To establish animal model of osteoporosis of the male rats, to observed the effect of Yougui pill on bone metabolism in experimental rat with osteoporosis, and to explore Yougui pills'effect of inhibiting rat alveolar bone absorption. For further understand the role of traditional Chinese medicine for prevention and treatment alveolar bone absorption in patients with osteoporosis, especially gerontism male to provide an experimental reference.Methods: Fifty 6-month-old male Wistar rats were randomly divided into five groups: normal group, hormone group(group model 1), orchidectomy group(group model 2), hormone with Yougui pill therapy group(group THER 1), orchidectomy with Yougui pill therapy group(group THER 2) and each group 10. Group ORX, group ORX-YGT to get dorsal position removal didymus under general anaesthesia with 10% KOD. Suture nick, peritoneal injection GM 1ml. 4 weeks after orchidectomized, rats of group H, group H-YGT drench Hydrocortisone injection, 3.6mg/kg, 6 times per week for the first 30 days, 2 times per week for the later 30 days, 2 months in all. After 3 months of orchidectomy, draw-off animal randomly, the maxilla and femur tissue sections were stained with HE for histological examination. Osteoporosis model identification is proved to be successful, two therapy groups rats were treated with Yougui pills, each rat gavage daily from 2ml(5mg/kg), the remaining three groups were treated with oral administration of normal saline 2ml water daily. After 3 months, determine femoral bone BMD, then all rats were bled to death by femoral arteries bledding, collecting whole blood for serum T and E2 with radioimmunity; collected maxillae and femoral bone, and was immediately put into 10% formaldehyde buffer solution for fixation, after 24h, changed to 10%EDTA decalcification solution 2 months, the tissue sections were stained with HE for histological examination; and the TGF-β1 expression were detected by immunohistochemical method.Results:1 A week after modeling, the rats exhibited weight loss, hair thinning, fatigue less dynamic, chills and crowding together, eat and drink less, and the urination frequency and volume of urine increased "deficiency of kidney-YANG" symptoms. 3 months after orchidectomy, body weight slower growth in hormone group (P<0.01), while the ORX group gained weight faster (P<0.01). Model and normal group rats were randomly sampled, take the maxillae and femur and the animals were killed, making biopsy section, trabecular bone thick, dense in the normal group; model group exhibited femoral trabecular bone loose, arrangement of disorder. Indicates that the two methods has been successfully in establishing animal models of osteoporosis. After 3 months Yougui Pill treatment, the hormone group body weight was significantly lower than normal group (P<0.01), body weight of orchidectomized rats was significantly higher than the normal group (P<0.01); group H-YGT weight higher than the hormone group (P<0.05), group ORX-YGT reduced body weight compared with ORX group (P<0.01), the weight of two treatment groups rats compared with group normal were no significant change (P>0.05), and between the two treatment groups there was no significant difference (P> 0.05).2 After 3 months Yougui Pill treatment, the rats femur bone mineral density among five groups were compared. Two model group was significantly lower than group normal(P<0.01), and no significant difference between the two groups (P>0.05); two treatment group was significantly higher than the model group(P<0.01) and lower than group normal(P<0.01), between the two treatment groups was no significant difference (P>0.05).3 Radioimmunoassay determination of serum E2 and T, the E2 concentration in rats was no significant difference (P>0.05). The T content of two model group was significantly lower than group normal (P<0.01), and no significant difference between the two groups (P>0.05); group H-YGT was significantly higher than the group hormone (P<0.01), and higher than the group normal (P<0.05); group ORX-YGT and group ORX no significant difference (P>0.05), lower than the group normal (P<0.01); group H-YGT was significantly higher than group ORX-YGT , there was significant difference (P<0.01).4 Rat maxillae and femur tissue HE staining and bone histomorphometry parameters were measured.Three months after medication, morphology observation: normal femoral trabecular was thick, orderly, cis-arranged by the rules of force direction; maxillae has no absorption, which is fraught with bone marrow, trabecular bone was flaky, enlargement. Model group femoral trabecular was smaller and trabecular spaces scale-up, resulting the bone marrow cavity expansion; maxillae resorption, trabecular thinning, alveolar bone surface has absorption lacuna and osteoclasts. Two treatment groups has larger number of femoral trabecular, arranged more closely, close to the group normal, no significant expansion of the bone marrow cavity; maxillae trabecular bone thicker and trabecular spaces decreases, which is fraught with red bone marrow , absorption lacuna more osteoblasts can be seen.Bone histomorphometry parameters were measured: two model group, maxillae and femur Tb.Ar(%),Tb.Th(μm) were significantly lower than the group normal(P<0.01), the Tb.Sp(μm)was significantly higher than group normal (P<0.01), but between the two groups no significant difference (P>0.05); two treatment group Tb.Ar(%),Tb.Th(μm) were significantly higher than the model group(P<0.01), the Tb.Sp(μm)was significantly lower than model group(P<0.01), the two treatment groups has no significant difference (P>0.05).5 TGF-β1 expression in maxillae, in group normal, cavitas medullaris, osteoblasts showed positive expression, some bone matrix and bone cells can also be stainned; in model group, cavitas medullaris revealed weakly positive expression, osteogenic cells has no obvious expression; in treatment group, cavitas medullaris, osteoblasts showed strong positive expression, in some vascular endothelial cells, bone cells and bone matrix can also be found positive expression.6 TGF-β1 immunohistochemistry mean optical density (OD) measured, group hormone and group ORX was significantly lower than group normal(P <0.01), no difference between the two groups (P> 0.05); treatment group was significantly higher than the model group (P <0.01), and the two treatment groups no significant difference (P> 0.05).Conclusion: 1 Hydrocortisone and orchidectomized successfully established rat model of osteoporosis. 2 When all over the body occurrence osteoporosis, the jaw bones also can be occurrenced osteoporosis at the same time, but the jaw bone resorption is less than the femur bone resorption. 3 Yougui pill slowed down osteoporosis occurrence due to lower testosterone levels, slowed down the jaw bone absorption, and promoted new bone formation, improved the alveolar bone structure, indicates it has certain therapeutic effect on jaw bone absorb caused by osteoporosis. 4 Yougui pill can increased sex hormones level; has androgen-like effect. 5 Yougui pill can increased TGF-β1 expression in jaw bone tissue, induction bone formation improve the jaw bone structure. |
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