Anti-osteoporosis effects of Anemarrhenae Rhizoma / Phellodendri Chinensis Cortex herb pair and its major active components in diabetic rats and zebrafish.
Anti-osteoporosis effects of Anemarrhenae Rhizoma / Phellodendri Chinensis Cortex herb pair and its major active components in diabetic rats and zebrafish.
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DOI：
10.1016/j.jep.2022.115269
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发表时间：
2022-04
影响因子：
5.4
通讯作者：
Pingcui Xu;Bingfeng Lin;Xuehui Deng;Shiwei He;Ning Chen;Nani Wang
中科院分区：
文献类型：
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作者：
Pingcui Xu;Bingfeng Lin;Xuehui Deng;Shiwei He;Ning Chen;Nani Wang
民族药理学相关性当归/陈皮（AR/PCC）药对已被广泛应用于治疗糖尿病性骨质疏松症的中药中。然而，AR/PCC的抗糖尿病代谢活性成分仍不清楚。材料与方法制备不同比例（AR：PCC = 1：3，1：2，1：1，2：1，3：1，w/w）的AR/PCC水提物，并将水提物与水提物按一定比例混合，制备水提物。链脲佐菌素诱导的糖尿病大鼠口服给予AR/PCC提取物。采用超高效液相色谱-四极杆飞行时间质谱法对糖尿病大鼠血清中吸收的植物化学成分进行了鉴定，并采用高效液相色谱-紫外检测器-蒸发光散射检测器法测定了AR/PCC提取物中的含量。提取物中的吸附物被认为是AR/PCC的主要潜在活性成分，它们的组合被定义为M-AR/PCC。采用组分敲除法评价各化合物在M-AR/PCC中的作用。然后使用糖尿病斑马鱼模型的幼体和成体来评价M-AR/PCC的抗糖尿病代谢性能。结果口服不同比例的AR/PCC提取物可逆转链脲佐菌素（STZ）诱导的糖尿病性骨质疏松大鼠的骨密度、骨体积密度、骨小梁厚度、骨小梁数量、骨密度减少股骨干骺端骨小梁分离。在血清中检测到7种植物化学成分，其中包括1种氧杂蒽酮苷和6种生物碱。利用糖尿病斑马鱼幼鱼模型和化合物敲除策略，证明M-AR/PCC中的各个化合物在糖尿病斑马鱼骨量的正向调节作用中发挥着不可或缺的作用。以1：1的中药组合及相应的M-AR/PCC对糖尿病骨质疏松症的治疗效果最好。在糖尿病成年斑马鱼模型中，它们对抗酒石酸酸性磷酸酶活性的抑制作用和对碱性磷酸酶活性的促进作用相似。结论AR/PCC中药对及其主要活性成分对链脲佐菌素诱导的斑马鱼糖尿病模型具有明显的抗糖尿病作用。来自AR/PCC草药对的七种活性化合物的组合可能是预防糖尿病相关骨质疏松症的潜在药物。
Ethnopharmacological relevanceAnemarrhenae Rhizoma/Phellodendri Chinensis Cortex (AR/PCC) herb pair has been widely used in traditional Chinese medicines for the treatment of diabetic osteoporosis. However, the anti-diabetic osteoporotic active components of AR/PCC remain unclear. This study aimed to explore the major active ingredients in AR/PCC for its protective effects against bone deterioration induced by diabetes.Materials and methodsThe aqueous extracts of AR/PCC with different proportions (AR:PCC = 1:3, 1:2, 1:1, 2:1 and 3:1, w/w) were prepared. Streptozotocin-induced diabetic rats were orally administrated with the AR/PCC extracts. The absorbed phytochemical compounds in serum of diabetic rats were identified by ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry method and their contents in the AR/PCC extracts were determined by high performance liquid chromatography-ultraviolet detector-evaporative light scattering detector method. The absorbed compounds in the extracts were considered as the major potential active components in AR/PCC, and their combination was defined as M-AR/PCC. A component-knockout approach was applied to evaluate the contribution of each compound in M-AR/PCC. The larvae and adults of diabetic zebrafish models were then used to evaluated the anti-diabetic osteoporotic performance of the M-AR/PCC. The real-time reverse transcription polymerase chain reaction technique was applied to study the regulation effects of M-AR/PCC on osteogenesis and osteoclastgensis in diabetic zebrafish models.ResultsThe phenotypes of diabetic osteoporosis rats induced by streptozotocin were reversed by the oral administration of AR/PCC extracts with different ratios, as evidenced by the increased bone mineral density, bone volume density, trabecular thickness, trabecular number, and decreased trabecular separation of femoral metaphysis. Seven phytochemical compounds were detected in the serum and their contents in AR/PCC varied dramatically with different proportions, including 1 xanthone glycoside and 6 alkaloids. By using diabetic zebrafish larvae model and compound-knockout strategy, each compound in M-AR/PCC were proved to play an indispensable role in the positive regulatory actions in the bone mass of diabetic zebrafish. Furthermore, the herb pair with a ratio of 1:1 and the related M-AR/PCC showed the best therapeutic effects on diabetic osteoporosis. They showed similar performances on the inhibition of the tartrate-resistant acid phosphatase activity and the promotion of the alkaline phosphatase activity in diabetic adult zebrafish model. The M-AR/PCC treatment could decrease the blood glucose, upregulate the mRNA expression levels of osteoblast-related genes (alp, runx2bandopg) and downregulate the expression of osteoclast-related genes (acp5α, ranklandsost) in streptozotocin-induced zebrafish.ConclusionAR/PCC herb pair and its major active components possess potent anti-diabetic osteoporotic effect on streptozotocin-inducedin vivomodels. The combination of the seven active compounds derived from AR/PCC herbal pair could be a potential agent for protection against osteoporosis associated with diabetes.