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| 由Th17/Treg细胞免疫平衡探讨金刚丸对去势大鼠的骨保护机制 |
| Exploring the bone protective mechanism of Jingang Pills on OVX rats based on Th17/Treg cell immune balance |
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| DOI：10.3969/j.issn.1006-7108.2026.01.002 |
| 中文关键词: 金刚丸 骨质疏松 Th17/Treg 免疫 骨丢失 |
| 英文关键词:Jingang Pills osteoporosis Th17/Treg immunity bone loss |
| 基金项目:国家自然科学基金（82405447）；湖南省中医药管理局重点项目（A2024012）；湖南省研究生科研创新项目（2024CX068，CX20240736） |
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| 中文摘要: |
| 目的 从Th17/Treg细胞免疫平衡的角度探讨温补肾阳方药金刚丸干预绝经后骨质疏松症的作用机制。方法 40只雌性SD大鼠按随机数字表法分为假手术组、模型组、戊酸雌二醇组[0.1mg/(kg·d)]、金刚丸等效剂量组[3.6g/(kg·d)]、金刚丸高剂量组[7.2g/(kg·d)]，各8只，采用双侧卵巢摘除术造模。药物干预3个月后收集大鼠双侧股骨、脾脏及血液，采用Mirco-CT检测骨微结构，抗酒石酸酸性磷酸酶染色（TRAP）观察破骨细胞数量，酶联免疫吸附法(ELISA)检测血清中IL-17、IL-10水平，流式细胞术检测大鼠血液和脾脏的Th17/Treg细胞比例，实时荧光定量聚合酶链式反应法（RT-qPCR）和蛋白免疫印迹（WB）分别检测骨组织ROR-γt、Foxp3的mRNA和蛋白表达。结果 与模型组比较，金刚丸能显著提高去势大鼠骨组织BMD、BV/TV、Tb.N（P＜0.01），降低Tb.SP（P＜0.01），改善骨小梁结构；能有效降低去势大鼠骨组织中破骨细胞数量；能显著降低去势大鼠血清IL-17水平（P＜0.01），提高IL-10水平（P＜0.01）；能显著降低去势大鼠血液和脾脏CD4+IL-17A（Th17）比率（P＜0.01），提高CD4+CD25+Foxp3（Treg）比率（P＜0.01）；并显著降低股骨中ROR-γt的mRNA和蛋白水平（P＜0.01），提高Foxp3的mRNA和蛋白水平（P＜0.01）。结论 金刚丸可通过抑制破骨细胞活性而有效改善去势大鼠的骨丢失，其作用机制与调节Th17/Treg细胞平衡密切相关。 |
| 英文摘要: |
| Objective To explore the mechanism of action of the warming and tonifying kidney-yang prescription Jingang Pills in intervening postmenopausal osteoporosis from the perspective of Th17/Treg cell immune balance. Methods Forty female SD rats were divided into the sham operation group, the model group, the estradiol valerate group (0.1 mg·kg?1·d?1), the equivalent-dose Jingang Pills group (3.6 g·kg?1·d?1), and the high-dose Jingang Pills group (7.2 g·kg?1·d?1) according to the random number table method, with 8 rats in each group. The bilateral ovariectomy was used to establish the model. After 3 months of drug intervention, the bilateral femurs, spleens and blood of the rats were collected. Micro-CT was used to detect the bone microstructure. Tartrate-resistant acid phosphatase staining (TRAP) was used to observe the number of osteoclasts. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of interleukin-17 (IL-17) and interleukin-10 (IL-10) in the serum. Flow cytometry was used to detect the proportion of Th17/Treg cells in the blood and spleen of rats. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blotting (WB) were respectively used to detect the mRNA and protein expressions of ROR-γt and Foxp3 in bone tissues. Results Compared to the model group, Jingang Pills can significantly increase the bone mineral density (BMD), bone volume fraction (BV/TV), and trabecular number (Tb.N) of bone tissues in ovariectomized rats (P＜0.01), decrease the trabecular separation (Tb.SP) (P＜0.01), and improve the trabecular structure. Meanwhile, Jingang Pills canreduce the number of osteoclasts in the bone tissues of ovariectomized rats. Additionally, Jingang Pills can significantly decrease the level of IL-17and increase the level of IL-10 in serum of ovariectomized rats (P＜0.01). Furthermore, Jingang Pills can significantly lower the ratio of CD4+IL-17A (Th17) in the blood and spleen of ovariectomized rats, and increase the ratio of CD4+CD25+Foxp3 (Treg) (P＜0.01). Moreover, Jingang Pills can significantly reduce the mRNA and protein expression levels of ROR-γt in the femur, and increase the mRNA and protein expression levels of Foxp3 (P＜0.01). Conclusion Jingang Pills can effectively reduce bone loss in ovariectomized rats by inhibiting the activity of osteoclasts, and its mechanism of action is closely related to the regulation of Th17/Treg cell balance. |
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