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目的通过观察狭鳕鱼皮胶原多肽对去势大鼠骨质疏松模型骨微结构的影响,探讨其防治骨质疏松的可行性。方法成年Wistar雌性大鼠60只,体质量(250±10)g,随机分为5组(n=12),分别为正常对照组(A组)、骨质疏松模型组(B组)、骨质疏松模型+狭鳕鱼皮胶原多肽预防组(C组)、骨质疏松模型+狭鳕鱼皮胶原多肽低剂量治疗组(D组)、骨质疏松模型+狭鳕鱼皮胶原多肽高剂量治疗组(E组),每组12只。B、C、D、E组采用摘除双侧卵巢法制备大鼠骨质疏松模型。C组从术前4周开始、D、E组从术后6周开始,每天按照1.0、0.5、1.0 g/kg行狭鳕鱼皮胶原多肽灌胃,连续6周;A、B组于术后同时间点给予等体积生理盐水灌胃。末次给药24 h后,A、B组大鼠摄股骨X线片并测定灰度值;然后颈椎脱臼法处死各组大鼠,取左侧胫骨近端骨组织行HE染色,观察骨组织病理学改变,测量骨小梁数量(trabecular number,TN)、平均骨小梁厚度(mean trabecular plate thickness,MTPT)、平均骨小梁间距(mean trabecular plate spacing,MTPS)、骨小梁体积百分比(trabecular bone volume,TBV)、平均骨皮质厚度(mean bone cortical thickness,MBCT);免疫组织化学染色观测降钙素受体(caltitonin receptor,CTR)及IL-1表达水平。结果 B组大鼠股骨灰度值显著低于A组(t=45.130,P=0.000),表明去势大鼠骨质疏松模型制备成功。组织学观察示,A、C、E组TN、MTPS、TBV、MBCT与B组比较,差异有统计学意义(P<0.05);C组各骨组织形态计量学参数与D、E组比较,差异均有统计学意义(P<0.05);D组TN、MTPS、TBV、MBCT与A组比较差异有统计学意义(P<0.05);E组仅MTPS与A组比较差异有统计学意义(P<0.05);E组MTPS、TBV、MBCT与D组比较,差异有统计学意义(P<0.05)。免疫组织化学染色观察示,A、C、D、E组CTR、IL-1表达水平较B组降低,C、E组低于D组,差异有统计学意义(P<0.05);其余组间比较差异均无统计学意义(P>0.05)。结论狭鳕鱼皮胶原多肽能够改善骨质疏松大鼠的骨微结构,其机制可能与抑制骨组织中CTR、IL-1表达有关,但尚未发现其对骨质疏松症有预防作用。
Objective To investigate the effect of Polypella sialis polypeptide on the microstructure of osteoporosis model in ovariectomized rats and its feasibility of prevention and treatment of osteoporosis. Methods Sixty adult Wistar female rats weighing 250 ± 10 g were randomly divided into 5 groups (n = 12), normal control group (group A), osteoporosis model group (group B), bone Osteoporosis model + low-dose cod-fish skin collagen peptide treatment group (D group), osteoporosis model + polio skin collagen peptide high-dose treatment group (group C) Group E), 12 in each group. B, C, D, E groups were divided into two groups by ovariectomized rat osteoporosis model. Groups C and D began to operate at 4 weeks preoperatively, and groups D and E started daily administration of 1.0, 0.5, and 1.0 g / kg of cod celled collagen polypeptide for 6 weeks after operation. Groups A and B received postoperative At the same time give equal volume of normal saline gavage. After the last administration for 24 h, the rats in groups A and B received the X-ray of the femur and measured the gray value. Then, the rats in each group were sacrificed by cervical dislocation, and the bone of the proximal tibia was removed by HE staining. The changes of physicochemical properties, trabecular number (TN), mean trabecular plate thickness (MTPT), mean trabecular plate spacing (MTPS), trabecular volume bone volume (TBV) and mean bone cortical thickness (MBCT). The expression of caltitonin receptor (CTR) and IL-1 were detected by immunohistochemical staining. Results The femoral gray value of group B was significantly lower than that of group A (t = 45.130, P = 0.000), indicating that osteoporosis model was successfully established in ovariectomized rats. Histological observation showed that TN, MTPS, TBV and MBCT in group A, C and E had statistical significance compared with group B (P <0.05). Compared with group D and group E, (P <0.05). The differences of TN, MTPS, TBV and MBCT between group D and group A were statistically significant (P <0.05), while the difference between group MT and group A was statistically significant P <0.05). The differences of MTPS, TBV, MBCT between group E and group D were statistically significant (P <0.05). Immunohistochemical staining showed that the expression of CTR and IL-1 in group A, C, D and E were lower than those in group B, while those in group C and E were lower than those in group D (P <0.05) There was no significant difference between the two groups (P> 0.05). Conclusion Polypeptide codon skin collagen polypeptide can improve the bone micro-structure of osteoporosis rats. Its mechanism may be related to the inhibition of the expression of CTR and IL-1 in bone tissue. However, it has not been found that it has a preventive effect on osteoporosis.