银杏叶提取物促进大鼠激素性股骨头坏死中H亚型微血管形成的实验研究

Experimental study on the promotion of H-type microvessel formation in hormone-induced femoral head necrosis in rats by Ginkgo biloba leaf extract

摘要:

目的 观察银杏叶提取物在大鼠激素性股骨头坏死（SONFH）模型中对H亚型微血管的影响。方法 采用脂多糖联合甲泼尼龙法复制SONFH模型。将50只大鼠分为空白组、模型组，以及银杏叶提取物高、中、低剂量组，每组10只。银杏叶提取物高、中、低剂量组分别予0.88、0.43、0.21 mL/kg银杏叶提取物溶液尾静脉注射，空白组和模型组分别予等量生理盐水尾静脉注射。4周时Micro-CT断层扫描和HE染色观察股骨头外形、骨小梁结构及囊性变情况；治疗8周后，采用HE染色观察检测股骨头软骨表面光滑度和骨小梁密度，免疫荧光染色观察检测股骨头血小板-内皮细胞黏附分子（CD31）和内皮黏蛋白（Emcn）表达。结果 4周后Micro-CT断层扫描可见大鼠股骨头变形，骨小梁稀疏；HE染色结果可见骨小梁稀疏、断裂、股骨头坏死，Micro-CT断层扫描与HE染色结果一致，表明大鼠SONFH模型复制成功。8周后HE染色结果显示，与空白组相比，银杏叶提取物高、中、低剂量组及模型组均有不同程度股骨头坏死，但银杏叶提取物高、中、低剂量组与模型组比较，骨小梁相对密集，股骨头坏死减少。各组大鼠CD31及Emcn阳性率比较，差异均有统计学意义（P <0.05），模型组CD31和Emcn阳性率较空白组及银杏叶提取物高、中、低剂量组均降低（P <0.05），银杏叶提取物高剂量组CD31阳性率与空白组比较，差异无统计学意义（P >0.05），但Emcn阳性率高于空白组（P <0.05），银杏叶提取物高剂量组CD31和Emcn阳性率与中、低剂量组比较，差异均有统计学意义（P <0.05），但中剂量与低剂量比较，差异无统计学意义（P >0.05）。结论 银杏叶提取物可促进大鼠SONFH模型中H亚型微血管形成，改善股骨头周围血循，促进血管成骨耦合，延缓疾病进程。

Abstract:

Objective To observe the effect of Ginkgo biloba leaf extract on H-type microvessel formation in a rat model of hormone-induced femoral head necrosis (SONFH).Methods SONFH model was established using lipopolysaccharide combined with methylprednisolone acetate in rats. Fifty rats were divided into blank group, model group, and high, medium, and low-dose Ginkgo biloba leaf extract groups, with 10 rats in each group. The high, medium, and low-dose Ginkgo biloba leaf extract groups were administered Ginkgo biloba leaf extract solution intravenously at doses of 0.88 mL/kg, 0.43 mL/kg, and 0.21 mL/kg, respectively, while the blank group and model group were given an equal volume of normal saline intravenously. After 4 weeks, micro-CT scanning and HE staining were used to observe the shape of the femoral head, trabecular structure, and cystic changes. After 8 weeks of treatment, HE staining was used to detect the smoothness of the cartilage surface of the femoral head and trabecular density, and immunofluorescence staining was used to detect the expression of platelet-endothelial cell adhesion molecule (CD31) and endomucin (Emcn) in the femoral head.Results Micro-CT scanning showed deformation of the femoral head and sparse trabeculae in rats after 4 weeks, and HE staining showed sparse trabeculae, fractures, and femoral head necrosis. The results of micro-CT scanning were consistent with HE staining, indicating successful replication of the SONFH model in rats. After 8 weeks, HE staining showed varying degrees of femoral head necrosis in the high, medium, and low-dose Ginkgo biloba leaf extract groups and the model group compared with the blank group. However, compared with the model group, the trabeculae were relatively dense, and femoral head necrosis was reduced in the high, medium, and low-dose Ginkgo biloba leaf extract groups. The positive rates of CD31 and Emcn in each group of rats were statistically significant (P < 0.05). The positive rates of CD31 and Emcn in the model group were lower than those in the blank group and the high, medium, and low-dose Ginkgo biloba leaf extract groups (P < 0.05). The positive rate of CD31 in the high-dose Ginkgo biloba leaf extract group was not statistically different from that in the blank group (P > 0.05), but the positive rate of Emcn was higher than that in the blank group (P < 0.05). The positive rates of CD31 and Emcn in the high-dose Ginkgo biloba leaf extract group were statistically different from those in the medium and low-dose groups (P < 0.05), but there was no significant difference between the medium and low-dose groups (P > 0.05).Conclusions Ginkgo biloba leaf extract can promote H-type microvessel formation in the SONFH model in rats, improve blood circulation around the femoral head, promote vascular-bone coupling, and delay disease progression.