特络细胞介导肥胖症大鼠膏摩寡肽中草药复合制剂的减肥作用及其机制

doi:10.12300/j.issn.1674-5817.2025.058

实验动物与比较医学 ›› 2025, Vol. 45 ›› Issue (5): 551-560.DOI: 10.12300/j.issn.1674-5817.2025.058

• 实验动物与中医药学专题 • 上一篇下一篇

特络细胞介导肥胖症大鼠膏摩寡肽中草药复合制剂的减肥作用及其机制

罗一凡¹(), 张臻玮¹, 梅璐¹, 史叶萍¹, 邢艺彤¹, 张泽奇²^,³^,⁴, 李楚欣²^,³^,⁴, 韩春霞²^,³^,⁴, 杨平顺²^,³^,⁴()(), 陈秋生¹()()

^1.南京农业大学动物医学院, 南京 210000
^2.深圳市容大生物科技股份有限公司, 深圳 518000
^3.东莞市容大生物科技有限公司, 东莞 523808
^4.广东省天然植物源先导化合物化妆品工程技术研究中心, 深圳 518000

收稿日期:2025-04-18 修回日期:2025-06-05 出版日期:2025-10-25 发布日期:2025-10-23
通讯作者:
陈秋生（1963—），男，博士，教授，博士生导师，主要从事经络实质细胞研究。E-mail： chenqsh305@njau.edu.cn。ORCID：0000-0002-6087-7029；

杨平顺（1967—），男，本科，高级工程师，主要从事化妆品研发。E-mail：524660751@qq.com。ORCID：0009-0007-6085-7937
作者简介:罗一凡（2000—），女，硕士研究生，主要从事经络实质细胞研究。E-mail： luoyifan0216@163.com。ORCID：0009-0005-0349-5429
基金资助:
深圳市容大生物科技股份有限公司-南京农业大学联合项目“基于激活特络细胞调节亚健康响应机制的方法探索与研究”(090HYQY23090)

Telocytes-Mediated Effects and Mechanisms of Anointing and Massage Therapy Using Oligopeptide-Herbal Medicine Composite Against Obesity in Rats

LUO Yifan¹(), ZHANG Zhenwei¹, MEI Lu¹, SHI Yeping¹, XING Yitong¹, ZHANG Zeqi²^,³^,⁴, LI Chuxin²^,³^,⁴, HAN Chunxia²^,³^,⁴, YANG Pingshun²^,³^,⁴()(), CHEN Qiusheng¹()()

^1.College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210000, China
^2.Shenzhen Rongda Biotechnology Co. , Ltd. , Shenzhen 518000, China
^3.Dongguan Rongda Biotechnology Co. , Ltd. , Dongguan 523808, China
^4.Guangdong Engineering Research Center of Natural Plant-Derived Lead Compounds for Cosmetics, Shenzhen 518000, China

Received:2025-04-18 Revised:2025-06-05 Published:2025-10-25 Online:2025-10-23
Contact: CHEN Qiusheng (ORCID: 0000-0002-6087-7029), E-mail: chenqsh305@njau.edu.cn;
YANG Pingshun (ORCID: 0009-0007-6085-7937), E-mail: 524660751@qq.com

摘要/Abstract

摘要：

目的探讨寡肽中草药复合制剂的减肥功效及其机制，为肥胖治疗提供新思路。方法将23只SPF级雌性SD大鼠随机分为对照组（3只）和造模组（20只），对照组连续6周饲喂普通饲料，造模组连续6周高脂饮食诱导构建单纯性肥胖大鼠模型。造模成功后，随机将造模组分为模型组（10只）和治疗组（10只），治疗组开始通过膏摩寡肽中草药复合制剂（3 g/只/次，1天1次，1次20 min）记作第1天，对照组和模型组不作寡肽中草药复合制剂处理但施以相同的按摩处理，并分别继续饲喂普通饲料和高脂饲料。将模型组和治疗组各10只大鼠分别分为3组，每组为3、3、4只，分别于治疗后第3、6、9天将3组大鼠经异氟烷呼吸麻醉后称重并行心脏采血，继续心脏放血法安乐死后采集腹部皮下脂肪组织（abdominal subcutaneous adipose tissue，aSAT）和肾周白色脂肪组织（perirenal white adipose tissue，pWAT）样本并称重。检测血清甘油三酯（triglycerides，TG）和高密度脂蛋白胆固醇（high-density lipoprotein cholesterol，HDL-C）水平。用Image J软件测量腹部皮下脂肪组织厚度、肾周白色脂肪细胞以及腹部皮下脂肪细胞直径和面积；通过免疫荧光技术观察腹部皮下脂肪组织单位面积的特络细胞（telocytes，TCs）、细胞连接、外泌体数量；通过透射电子显微镜统计腹部皮肤真皮及皮下TCs突起（telopodes，Tps）长度，并观测外泌体囊泡、粗面内质网、线粒体、细胞连接的分布。结果与对照组相比，造模组大鼠的体重显著增加（P<0.05）。与对照组相比，模型组大鼠血清TG水平显著升高，HDL-C水平显著降低，肾周白色脂肪组织质量显著增加（均P<0.05）。与模型组相比，治疗组大鼠的体重下降率显著增高，肾周白色脂肪组织质量显著降低，血清TG水平显著降低，HDL-C水平显著升高（均P<0.05）。与模型组相比，治疗组腹部皮下脂肪组织厚度、腹部皮下脂肪与肾周白色脂肪组织的细胞直径和面积显著减小（均P<0.05）。与模型组相比，治疗组腹部皮下脂肪组织单位面积TCs数量、腹部皮下脂肪组织TCs外泌体数量、Tps长度显著增加（均P<0.05）；治疗组TCs细胞内线粒体、粗面内质网，细胞连接数均有增多趋势。结论膏摩寡肽中草药复合制剂通过调节TCs及其细胞间通讯，有效改善肥胖大鼠的血脂紊乱和脂肪堆积等肥胖相关症状。

关键词: 特络细胞, 寡肽中草药复合制剂, 肥胖, 大鼠, 膏摩

Abstract:

Objective To investigate the weight loss efficacy and mechanisms of oligopeptide-herbal medicine composite, and to provide new approaches for obesity treatment. Methods Twenty-three SPF female SD rats were randomly divided into control group (n=3) and modeling group (n=20). The control group was fed an ordinary diet for 6 consecutive weeks, and the modeling group was fed a high-fat diet for 6 consecutive weeks to establish a simple obesity rat model. After successful modeling, the modeling group was randomly divided into model group (n=10) and treatment group (n=10). The treatment group started anoint-and-massage therapy with oligopeptide-herbal medicine composite (3 g per rat per time,once a day, 20 min each time), which was recorded as day 1. The control group and model group were not treated with oligopeptide-herbal medicine composite but received the same massage, and continued to be fed ordinary feed and high-fat feed respectively. Ten rats in the model group and ten in the treatment group were each divided into 3 groups, with 3, 3, and 4 rats in each group. On days 3, 6, and 9 after treatment, the rats in the three groups were weighed and cardiac blood collection was performed after isoflurane respiratory anesthesia. After euthanasia by cardiac bloodletting, abdominal subcutaneous adipose tissue (aSAT) and perirenal white adipose tissue (pWAT) samples were collected and weighed. Serum triglycerides (TG) and high-density lipoprotein cholesterol (HDL-C) levels were measured. Image J software was used to measure aSAT thickness and the diameter and area of perirenal white adipocytes and abdominal subcutaneous adipocytes. Immunofluorescence technique was used to observe the number of telocytes (TCs), cell junctions and exosomes per unit area of aSAT. Transmission electron microscopy was used to measure the length of telopodes (Tps) of dermal and subcutaneous TCs in abdominal skin, and to observe the distribution of exosome vesicles, rough endoplasmic reticulum, mitochondria, and cell junctions. Results Compared with the control group, the body weight of rats in the modeling group increased significantly (P<0.05). Compared with the control group, rats in the model group showed a significant increase in serum TG level, a significant decrease in HDL-C level, and a significant increase in pWAT mass (all P<0.05). Compared with the model group, the treatment group showed a significantly increased rate of body weight reduction, significantly decreased pWAT mass, significantly decreased serum TG level, and significantly increased HDL-C level (all P<0.05). Compared with the model group, the treatment group showed significant reductions in aSAT thickness, as well as cell diameter and area in both aSAT and pWAT (all P<0.05). Compared with the model group, the treatment group showed significantly increased number of TCs per unit area of aSAT, number of exosomes from TCs in aSAT, and Tps length (all P<0.05). The treatment group showed an increasing trend in the numbers of mitochondria, rough endoplasmic reticulum, and cell junctions in TCs. Conclusion Oligopeptide-herbal medicine composite applied via anoint-and-massage therapy effectively improves obesity-related symptoms such as dyslipidemia and fat accumulation in obese rats by regulating TCs and their intercellular communication.

Key words: Telocytes, Oligopeptide-herbal medicine composite, Obesity, Rats, Anoint and massage

中图分类号:

Q95-33

罗一凡,张臻玮,梅璐,等. 特络细胞介导肥胖症大鼠膏摩寡肽中草药复合制剂的减肥作用及其机制[J]. 实验动物与比较医学, 2025, 45(5): 551-560. DOI: 10.12300/j.issn.1674-5817.2025.058.

LUO Yifan,ZHANG Zhenwei,MEI Lu,et al. Telocytes-Mediated Effects and Mechanisms of Anointing and Massage Therapy Using Oligopeptide-Herbal Medicine Composite Against Obesity in Rats[J]. Laboratory Animal and Comparative Medicine, 2025, 45(5): 551-560. DOI: 10.12300/j.issn.1674-5817.2025.058.

图/表 4

图1 大鼠高脂饮食诱导单纯性肥胖模型注：A，对照组（CG）和肥胖造模组（OG）大鼠在造模过程中的体重变化情况；B，对照组和模型组（MG）血清甘油三酯（TG）水平；C，对照组和模型组血清高密度脂蛋白胆固醇（HDL-C）水平；D~E，对照组和模型组大鼠肾周白色脂肪组织（pWAT）及其质量，其中D为代表性肾周白色脂肪组织对比图，E为对照组和模型组肾周白色脂肪组织质量。*P<0.05，**P<0.01，****P<0.000 1。

Figure 1 Simple obesity model induced by high-fat diet in ratsNote： A， Changes in body weight of rats in the control group （CG） and the obesity modeling group （OG） during the modeling process； B， Serum triglyceride （TG） levels in the control group and the model group （MG）； C， Serum high-density lipoprotein cholesterol （HDL-C） levels in the control group and the model group； D-E， Perirenal white adipose tissue （pWAT）and its weight of CG and MG rats， where the D are the representative comparison figures of perirenal white adipose tissue and E represents the perirenal white adipose tissue weight of CG and MG rats. *P<0.05， **P<0.01， ****P<0.000 1.

图2 膏摩寡肽中草药复合制剂加速大鼠体内脂质代谢注：A，模型组（MG）和治疗组（TMT）大鼠肾周白色脂肪组织（pWAT）；B，各组大鼠肾周白色脂肪组织质量变化情况；C，各组大鼠肾周白色脂肪组织HE染色；D，各组肾周脂肪细胞直径变化；E，各组肾周脂肪细胞面积变化；F，各组大鼠腹部皮下脂肪组织（aSAT）HE染色；G，各组腹部皮下脂肪厚度变化；H，各组腹部皮下脂肪细胞直径变化；I，各组腹部皮下脂肪细胞面积变化；J，各组大鼠血清甘油三酯（TG）变化情况；K，各组大鼠血清高密度脂蛋白胆固醇（HDL-C）变化情况；L，各组大鼠体重下降率变化情况。Day 3，Day 6，Day 9分别代表治疗第3、6、9天；*P<0.05，**P<0.01，***P<0.001，****P<0.000 1。C和F标尺为100 μm。

Figure 2 Anointing and massage with oligopeptide-herbal medicine composite accelerates lipid metabolism in ratsNote： A， Perirenal white adipose tissue （pWAT） of model group （MG） and treatment group （TMT） rats； B， Changes in perirenal white adipose tissue weight of model group and treatment group rats； C， HE staining of perirenal white adipose tissue of model group and treatment group rats； D， Changes in perirenal adipocyte diameter of each group； E， Changes in perirenal adipocyte area of each group； F， HE staining of abdominal subcutaneous adipose tissue （aSAT） of model group and treatment group rats； G， Changes in abdominal subcutaneous adipose tissue thickness of each group； H， Changes in abdominal subcutaneous adipocyte diameter of each group； I， Changes in abdominal subcutaneous adipocyte area of each group； J， Changes in serum triglyceride （TG） of model group and treatment group rats； K， Changes in serum high-density lipoprotein cholesterol （HDL-C） of model group and treatment group rats； L， Changes in body-weight loss rate of model group and treatment group rats. Day 3， Day 6， Day 9 represent the 3rd， 6th， 9th day of treatment， respectively； *P<0.05， **P<0.01， ***P<0.001， ****P<0.000 1. Scale bars in C and F indicate 100 μm.

图3 膏摩寡肽中草药复合制剂对大鼠特络细胞的影响注：A，模型组（MG）和治疗组（TMT）大鼠腹部皮下脂肪（A1、A2）和肾周白色脂肪（A3、A4）中特络细胞（TCs）（白色箭头所指）的免疫荧光表征；B，模型组和治疗组单位面积腹部皮下脂肪中的TCs数量变化；C，模型组和治疗组肾周白色脂肪中的TCs数量变化；D，透射电子显微镜图片中模型组和治疗组大鼠腹部皮下脂肪组织的TCs突起（Tps）（黑色箭头所指）；E，模型组和治疗组TCs的Tps长度；*P<0.05，**P<0.01；A图标尺为50 μm，D图标尺为2 μm。

Figure 3 Effect of anointing and massage with oligopeptide-herbal medicine composite on telocytesNote： A， Immunofluorescence characterization of telocytes （TCs， indicated by the white arrows） in abdominal subcutaneous adipose tissue （A1， A2） and perirenal white adipose tissue （A3， A4） of model group （MG） and treatment group （TMT） rats； B， Changes in the number of TCs per unit area of abdominal subcutaneous adipose tissue of the model group and the treatment group； C， Changes in the number of TCs in perirenal white adipose tissue of the model group and the treatment group； D， Telopodes （Tps， indicated by the black arrows） in abdominal subcutaneous adipose tissue of the model group and the treatment group rats as shown in the transmission electron microscope images； E， Tps length of TCs in the model group and the treatment group； *P<0.05， **P<0.01； Scale bar in A indicates 50 μm and in D indicates 2 μm.

图4 膏摩寡肽中草药复合制剂对大鼠特洛细胞的亚细胞器和细胞间连接的影响注：A，免疫荧光结果图展示的模型组（MG）和治疗组（TMT）大鼠肾周白色脂肪组织（第一行）与腹部皮下脂肪组织（第二行）中间隙连接蛋白43（Cx43）标记的缝隙连接（白色箭头所指）；B，透射电子显微镜下的特洛细胞（TCs，白色三角形所示）与外泌体囊泡（黑色三角形）、线粒体（黑色波浪箭头）、细胞连接（黑色箭头）和粗面内质网（白色五角星）；C，模型组和治疗组大鼠腹部皮下脂肪组织中TCs分泌外泌体数量的比较；*P<0.05；A图标尺为50 μm，B图标尺为1 μm。

Figure 4 Effect of anointing and massage with oligopeptide-herbal medicine composite on subcellular organelles and intercellular junctions of telocytesNote： A， Immunofluorescence images show the gap junctions （indicated by white arrows） labeled with connexin 43 （Cx43） in the perirenal white adipose tissue （the first row） and the abdominal subcutaneous adipose tissue （the second row） of model group （MG） and treatment group （TMT） rats； B， Transmission electron micrographs of TCs （white triangles） and exosome vesicles （black triangles）， mitochondria （black wavy arrows）， cell junctions （black arrows） and rough endoplasmic reticulum （white five-pointed star）； C， The number comparison of exosomes secreted by TCs in abdominal subcutaneous adipose tissue between the model and treatment groups； *P<0.05； Scale bar in A indicates 50 μm and in B indicates 1 μm.

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特络细胞介导肥胖症大鼠膏摩寡肽中草药复合制剂的减肥作用及其机制

Telocytes-Mediated Effects and Mechanisms of Anointing and Massage Therapy Using Oligopeptide-Herbal Medicine Composite Against Obesity in Rats

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