荜茇（Piperis longi fructus）在中药和民族药中应用广泛。在民族药尤其是蒙药中主要用于祛“巴达干·赫依”和降血脂；而在中药传统应用中，具有“温中散寒、下气止痛”功效。且两种理论体系下用药形式具有差异，其在民族药中多以生药形式打粉入药，而在传统中医药中则为煎煮入药。目前虽然已有荜茇在改善糖脂代谢紊乱等方面的研究，但其在双医药理论下发挥不同药效的物质基础和作用机制尚不明确。本研究旨在系统地评估荜茇改善糖脂代谢紊乱的民族药特色功效和改善胃寒及镇痛的中医药传统功效，解析其在双医药理论体系下药效差异的物质基础，探究其发挥不同功效的作用机制，为荜茇的临床应用提供实验依据和数据支撑。

在荜茇改善肥胖脂代谢紊乱的作用及机制研究中，基于油酸诱导的HepG2细胞、棕榈酸诱导的H9c2细胞脂质累积模型和3T3-L1前脂肪细胞诱导分化成熟模型，采用油红和Bodipy染色对荜茇醇提物、水提物、醇提物中的胡椒碱、非胡椒碱成分及6个单体的降脂活性进行评估，并通过实时荧光定量聚合酶链式反应（RT-qPCR）和转录组学技术初步分析有效成分的作用机制。结果发现，荜茇醇提物可抑制三种细胞模型脂质累积，而从醇提物中分离出的单体胡椒碱、PL17、PL13、PL3均可抑制3T3-L1细胞分化成熟，药效强度表现为PL17＞胡椒碱＞PL3＞PL13。RT-qPCR结果表明，荜茇醇提物可下调3T3-L1细胞的脂肪分化与合成基因（Ppar-γ、Rxr-α、Cebp-α、Fabp4、Fasn）、上调脂质分解基因（Atgl）的mRNA相对表达量。测序结果表明PL17可下调3T3-L1细胞的细胞呼吸链相关基因（Cycs、Ndufv1、Atp5al）和脂肪分化与合成基因（Dgat2、Hacd2、Cebp-α、Fabp4、Adipoq）、上调脂质β氧化基因（Cpt-1α、Acsl3、Acsbg1）的mRNA表达。基于荜茇醇提物在细胞实验中具有较好的降脂活性，随后采用ob/ob自发性肥胖小鼠模型和高脂饮食（HFD）诱导的肥胖小鼠模型进行荜茇醇提物和粗粉改善脂代谢紊乱作用的体内验证。通过生理指标（体重、体脂、肛温、进食量、呼吸代谢等）、生化指标（血脂、肝脂、血糖、炎症因子等）、葡萄糖稳态（葡萄糖耐量、胰岛素耐量）以及组织形态学变化等评估药效。结果发现荜茇醇提物对ob/ob小鼠无显著改善作用；荜茇粗粉可显著改善HFD诱导肥胖小鼠体重、体脂、空腹血糖和炎症因子（TNF-α）水平，荜茇醇提物改善小鼠空腹血糖和炎症因子（TNF-α）水平。

在荜茇改善糖尿病糖脂代谢紊乱的作用及机制研究中，通过构建棕榈酸（PA）和地塞米松（DEX）诱导的HepG2细胞胰岛素抵抗模型，以细胞葡萄糖耗量为指标评价荜茇醇提物和水提物药效，结果显示二者均可改善DEX诱导的HepG2细胞胰岛素抵抗，且醇提物药效优于水提物，但对PA诱导的模型无显著作用。进一步检测α-葡萄糖苷酶抑制活性、糖代谢及铁死亡相关基因mRNA表达情况，对活性成分的作用机制进行初步探讨。结果表明，荜茇醇提物和水提物对α-葡萄糖苷酶和铁死亡通路均无显著影响；但荜茇水提物可显著上调糖代谢相关基因mRNA水平（INSR、IRS1和G6P）和抗氧化酶NQO1基因mRNA的表达。基于两种提取物在细胞实验中均具有较好的改善胰岛素抵抗作用，随后采用db/db自发性糖尿病小鼠模型对荜茇进行体内药效验证。通过检测生理指标（进食量、饮水量、尿量、体重、体脂、肛温等）、生化指标（空腹血糖、胰岛素、糖化血红蛋白、尿糖、血脂、肝脂、炎症因子等）和葡萄糖稳态（糖耐量、胰岛素耐量）等，对荜茇粗粉、荜茇95%乙醇提取物、荜茇50%乙醇提取物、荜茇水提物及荜茇水煎液进行药效评估。结果发现荜茇粗粉可降低db/db小鼠的空腹血糖、炎症因子（IL-6、TNF-α）并改善糖耐量和胰岛素耐量，而荜茇醇提物、水提物和荜茇水煎液可改善糖耐量（30 min）及炎症因子（IL-1β、IL-6）水平。

采用热板实验和醋酸扭体实验评价荜茇的镇痛作用。通过小鼠痛阈、扭体潜伏期、扭体次数等指标评估荜茇粗粉、荜茇水煎液、荜茇醇提物的镇痛药效。结果发现热板实验中各给药组对小鼠痛阈无显著影响；醋酸扭体实验中，荜茇粗粉、水煎液和醇提物均显著减少小鼠醋酸扭体次数，荜茇水煎液还显著延长小鼠扭体潜伏期。

采用冰水灌胃联合冷水浴建立胃寒证模型探究荜茇改善胃寒的作用及机制。通过检测小鼠体重、肛温、胃内残留率、小肠推进率、肝糖原含量等评估荜茇粗粉、荜茇水煎液、荜茇醇提物改善胃寒的药效，结果表明荜茇粗粉、醇提物可显著提高胃寒证小鼠肛温、降低体脂比、增加瘦肉比；荜茇粗粉还显著降低胃内残留率、改善肝脏质量下降并增加肝糖原储存；荜茇水煎液未见显著药效。采用RT-qPCR检测炎症和产热基因表达情况对荜茇的作用机制进行初步探究，结果表明荜茇醇提物显著上调胃组织产热基因Dio2的mRNA相对表达量。

综上所述，本研究对荜茇的民族药特色功效及中医药传统功效展开系统探究。在民族药特色功效方面，荜茇醇提物抑制体外模型脂质累积和脂肪细胞分化、醇提物和水提物改善细胞模型胰岛素抵抗；动物实验结果表明荜茇粗粉显著改善肥胖及糖尿病糖脂代谢紊乱。该部分结果验证了荜茇以生粉入药为民族医药理论体系下发挥药效的最佳用药形式。在中医药传统功效研究中，体内实验表明荜茇粗粉、水煎液和醇提物可减轻小鼠的醋酸致扭体疼痛，且荜茇粗粉、醇提物可改善小鼠的胃寒证。该部分结果验证了荜茇以生粉、煎煮和提取物入药为中医药理论体系下发挥药效的主要用药形式。因此，不同用药形式可能是荜茇在双医药理论体系下发挥差异功效的重要因素。通过进一步挖掘多种入药形式下活性成分和分子机制差异，可为荜茇在双医药理论体系下同药不同效的临床应用提供药效物质基础和作用机制的实验依据。

Piperis longi fructus is widely used in traditional Chinese medicine and ethnic medicine. In ethnic medicine, especially in Mongolian medicine, it is mainly used to dispel "Bada gan·Heyi" and reduce blood lipids; while in the traditional application of traditional Chinese medicine, it has the effects of "warming the middle-jiao to dispel cold, descending qi to relieve pain". Moreover, the forms of medication under the two theoretical systems are different. In ethnic medicine, it is mostly used as raw medicine in the form of powder, while in traditional Chinese medicine, it is decocted for use. At present, although there are studies on Piperis longi fructus in improving disorders of glucose and lipid metabolism, the material basis and action mechanisms of its different pharmacological effects under the two medical theories are still unclear. This study aims to systematically evaluate the characteristic efficacy of ethnic medicine of Piperis longi fructus in improving disorders of glucose and lipid metabolism, as well as the traditional efficacy of traditional Chinese medicine in improving stomach cold and relieving pain, analyze the material basis of the differences in its pharmacological effects under the two medical theoretical systems, explore the action mechanisms of its different effects, and provide experimental evidence and data support for the clinical application of Piperis longi fructus.

In the study on the effect and mechanism of Piperis longi fructus in improving lipid metabolism disorder in obesity, based on the lipid accumulation models of HepG2 cells induced by oleic acid, H9c2 cells induced by palmitic acid, and the model of 3T3-L1 pre-adipocytes induced to differentiate into mature adipocytes, Oil Red and Bodipy staining were used to evaluate the lipid-lowering activities of the ethanol extract, water extract of Piperis longi fructus, piperine in the ethanol extract, non-piperine components, and six monomers. The action mechanisms of the effective components were preliminarily analyzed by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and transcriptomics technology. The results showed that the ethanol extract of Piperis longi fructus could inhibit the lipid accumulation in the three cell models. Among the monomers isolated from the ethanol extract, piperine, PL17, PL3, and PL13 could all inhibit the differentiation and maturation of 3T3-L1 cells, and the pharmacological effect intensity was PL17 > piperine > PL3 > PL13. The RT-qPCR results showed that the ethanol extract of Piperis longi fructus could down-regulate the relative mRNA expression levels of genes related to adipocyte differentiation and synthesis (Ppar-γ, Rxr-α, Cebp-α, Fabp4, Fasn) in 3T3-L1 cells and up-regulate the mRNA expression level of the lipid decomposition gene (Atgl). The sequencing results showed that PL17 could down-regulate the mRNA expression levels of genes related to the cellular respiratory chain (Cycs, Ndufv1, Atp5al) and genes related to adipocyte differentiation and synthesis (Dgat2, Hacd2, Cebp-α, Fabp4, Adipoq) in 3T3-L1 cells, and up-regulate the mRNA expression levels of genes related to lipid β-oxidation (Cpt-1α, Acsl3, Acsbg1). Based on the good lipid-lowering activity of the ethanol extract of Piperis longi fructus in cell experiments, the ob/ob spontaneous obese mouse model and the obese mouse model induced by a high-fat diet (HFD) were then used to verify the effect of the ethanol extract and crude powder of Piperis longi fructus in improving lipid metabolism disorder in vivo. The pharmacological effects were evaluated through physiological indexes (body weight, body fat, rectal temperature, food intake, respiratory metabolism, etc.), biochemical indexes (blood lipids, liver lipids, blood glucose, inflammatory factors, etc.), glucose homeostasis (glucose tolerance, insulin tolerance), and histomorphological changes. The results showed that the ethanol extract of Piperis longi fructus had no significant improvement effect on ob/ob mice; the crude powder of Piperis longi fructus could significantly improve the body weight, body fat, fasting blood glucose, and the level of inflammatory factor (TNF-α) in HFD-induced obese mice, and the ethanol extract of Piperis longi fructus could improve the fasting blood glucose and the level of inflammatory factor (TNF-α) in mice.

In the study on the effect and mechanism of Piperis longi fructus in improving glucose and lipid metabolism disorder in diabetes, by constructing the insulin resistance model of HepG2 cells induced by palmitic acid (PA) and dexamethasone (DEX), the pharmacological effects of the ethanol extract and water extract of Piperis longi fructus were evaluated using the cellular glucose consumption as an index. The results showed that both could improve the insulin resistance of HepG2 cells induced by DEX, and the pharmacological effect of the ethanol extract was better than that of the water extract, but there was no significant effect on the model induced by PA. The inhibitory activity of α-glucosidase, the expression of genes related to glucose metabolism and ferroptosis were further detected to preliminarily explore the action mechanism of the active components. The results showed that neither the ethanol extract nor the water extract of Piperis longi fructus had a significant effect on α-glucosidase and the ferroptosis pathway; however, the water extract of Piperis longi fructus could significantly up-regulate the mRNA levels of genes related to glucose metabolism (INSR, IRS1, and G6P) and the mRNA expression of the antioxidant enzyme NQO1 gene. Based on the good effect of the two extracts in improving insulin resistance in cell experiments, the db/db spontaneous diabetic mouse model was then used to verify the in vivo pharmacological effect of Piperis longi fructus. The pharmacological effects of the crude powder of Piperis longi fructus, the 95% ethanol extract of Piperis longi fructus, the 50% ethanol extract of Piperis longi fructus, the water extract of Piperis longi fructus, and the decoction of Piperis longi fructus were evaluated by detecting physiological indexes (food intake, water intake, urine volume, body weight, body fat, rectal temperature, etc.), biochemical indexes (fasting blood glucose, insulin, glycated hemoglobin, urine glucose, blood lipids, liver lipids, inflammatory factors, etc.), and glucose homeostasis (glucose tolerance, insulin tolerance). The results showed that the crude powder of Piperis longi fructus could reduce the fasting blood glucose and inflammatory factors (IL-6, TNF-α) in db/db mice and improve glucose tolerance and insulin tolerance, while the ethanol extract, water extract, and decoction of Piperis longi fructus could improve glucose tolerance (30 min) and the levels of inflammatory factors (IL-1β, IL-6).

The hot plate test and acetic acid-induced writhing test were used to evaluate the analgesic effect of Piperis longi fructus. The analgesic effects of the crude powder, decoction, and ethanol extract of Piperis longi fructus were evaluated through indexes such as the pain threshold of mice, writhing latency, and the number of writhing times. The results showed that each administration group had no significant effect on the pain threshold of mice in the hot plate test; in the acetic acid-induced writhing test, the crude powder, decoction, and ethanol extract of Piperis longi fructus could all significantly reduce the number of acetic acid-induced writhing times of mice, and the decoction of Piperis longi fructus could also significantly prolong the writhing latency of mice.

The model of stomach cold syndrome was established by intragastric administration of ice water combined with cold water bathing to explore the effect and mechanism of Piperis longi fructus in improving stomach cold. The effects of the crude powder, decoction, and ethanol extract of Piperis longi fructus in improving stomach cold were evaluated by detecting the body weight, rectal temperature, gastric residual rate, small intestine propulsion rate, and liver glycogen content of mice. The results showed that the crude powder and ethanol extract of Piperis longi fructus could significantly increase the rectal temperature of mice with stomach cold syndrome, reduce the body fat ratio, and increase the lean meat ratio; the crude powder of Piperis longi fructus could also significantly reduce the gastric residual rate, improve the decrease in liver quality, and increase liver glycogen storage; the decoction of Piperis longi fructus showed no significant pharmacological effect. RT-qPCR was used to detect the expression of inflammatory and thermogenic genes to preliminarily explore the action mechanism of Piperis longi fructus. The results showed that the ethanol extract of Piperis longi fructus could significantly up-regulate the relative mRNA expression level of the thermogenic gene Dio2 in the gastric tissue.

In conclusion, this study systematically explored the characteristic efficacy of ethnic medicine and the traditional efficacy of traditional Chinese medicine of Piperis longi fructus. In terms of the characteristic efficacy of ethnic medicine, the ethanol extract of Piperis longi fructus can inhibit lipid accumulation in in vitro models and the differentiation of adipocytes, and the ethanol extract and water extract can improve insulin resistance in cell models; the results of animal experiments show that the crude powder of Piperis longi fructus can significantly improve the disorders of glucose and lipid metabolism in obesity and diabetes. The results of this part verify that using the raw powder of Piperis longi fructus as medicine is the best form of medication for exerting its pharmacological effects under the theoretical system of ethnic medicine. In the study on the traditional efficacy of traditional Chinese medicine, in vivo experiments show that the crude powder, decoction, and ethanol extract of Piperis longi fructus can relieve the acetic acid-induced writhing pain in mice, and the crude powder and ethanol extract of Piperis longi fructus can improve the stomach cold syndrome in mice. The results of this part verify that using the raw powder, decoction, and extract of Piperis longi fructus as medicine are the main forms of medication for exerting its pharmacological effects under the theoretical system of traditional Chinese medicine. Therefore, different forms of medication may be an important factor for Piperis longi fructus to exert different effects under the two medical theoretical systems. By further exploring the differences in active components and molecular mechanisms under various forms of medication, it can provide experimental evidence of the pharmacological substance basis and action mechanism for the clinical application of Piperis longi fructus with different effects despite the same medicine under the two medical theoretical systems.