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Effect of herbal cake-partitioned moxibustion on Leptin/JAK2/STAT3 in lipid-lowering pathway of hyperlipidemia rabbits

隔药饼灸对高脂血症兔调脂通路Leptin/JAK2/STAT3的影响

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Abstract

Objective: To observe the lipid-lowering effect of different transdermal absorption enhancers applied to the herbal cake-partitioned moxibustion in hyperlipidemia model rabbits, and to explore the possible mechanism.

Methods: Forty New-Zealand rabbits were randomly divided into 5 groups using the random number table method, with 8 rats in each group. Rabbits in the blank group were fed routinely with normal diet; rabbits in the other groups were fed with high-fat diet for 12 weeks to establish the hyperlipidemia model. Rabbits in the blank and the model groups were not treated. After the model was prepared, rabbits in the non-transdermal absorption enhancer group received herbal cake-partitioned moxibustion without transdermal absorption enhancer; rabbits in the laurocapram group and the borneol group received herbal cake-partitioned moxibustion with laurocapram or borneol respectively. After 4 weeks of treatment, serum was collected for enzyme-linked immunosorbent assay (ELISA), and the liver tissues were isolated for immunohistochemistry, quantitative polymerase chain reaction (qPCR) and Western-blotting (WB) detection.

Results: Serum ELISA results showed that leptin was significantly decreased in the model group compared with the blank group (P<0.05); compared with the model group, leptin was significantly increased in the non-transdermal absorption enhancer, the laurocapram and the borneol groups (all P<0.05); compared with the non-transdermal absorption enhancer group, leptin was significantly increased in the laurocapram group and the borneol group (both P<0.05); there was no significant difference in leptin between the laurocapram and the borneol groups (P>0.05). The qPCR results of rabbit liver tissues showed that the mRNA expressions of leptin, Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) in the model group were significantly lower than those in the blank group (all P<0.05); compared with the model group, the mRNA expressions of leptin, leptin receptor (LR), JAK2 and STAT3 in the non-transdermal absorption enhancer, the laurocapram and the borneol groups were significantly increased (all P<0.05); compared with the non-transdermal absorption enhancer group, the mRNA expressions of leptin, LR, JAK2 and STAT3 in the laurocapram and the borneol groups were significantly increased (all P<0.05); compared with the laurocapram group, the mRNA expressions of leptin, LR, JAK2 and STAT3 in the borneol group were significantly increased (P<0.05). The trend of immunohistochemistry and WB detection results was basically consistent with the qPCR assay results. The immunohistochemistry and WB detection results of phosphorylated JAK2 (phospho-JAK2) and phosphorylated STAT3 (phospho-STAT3) were basically consistent with those of JAK2 and STAT3.

Conclusion: The molecular expression of Leptin/JAK2/STAT3 pathway in the hyperlipidemia model rabbits was decreased. The molecular expression of Leptin/JAK2/STAT3 pathway was significantly increased after the herbal cake-partitioned moxibustion. The application of laurocapram and borneol, as transdermal absorption enhancers, in the herbal cake-partitioned moxibustion could more obviously up-regulate the factors of the Leptin/JAK2/STAT3 lipid-regulating pathway than the herbal cake-partitioned moxibustion alone.

摘要

目的: 观察不同促透剂运用于隔药饼灸对高脂模型兔降脂效果的影响, 探讨其可能机制。

方法: 将40只 新西兰兔按随机数字表法随机分成5组, 每组8只。空白组正常喂养普通饲料; 其余组高脂饲料喂养12周, 复制高 脂模型。空白组和模型组不治疗。无促透剂组成模后采用不加促透剂药饼施灸; 氮酮组和冰片组分别将氮酮和冰 片用于药饼中, 进行隔药饼灸。治疗4周后, 分离血清进行酶联免疫吸附测定(ELISA), 分离肝脏组织进行免疫组 化、定量聚合酶链式反应(qPCR)和蛋白免疫印迹(WB)检测。

结果: 血清ELISA检测结果显示, 与空白组比较, 模型 组Leptin显著下降(P<0.05); 与模型组比较, 无促透剂组、氮酮组和冰片组Leptin显著升高(均P<0.05); 与无促透 剂组比较, 氮酮组和冰片组Leptin均显著升高(均P<0.05); 氮酮组和冰片组Leptin差异无统计学意义(P<0.05)。兔 肝脏组织qPCR结果显示, 与空白组比较, 模型组Leptin、JAK2和STAT3 mRNA表达显著降低(均P<0.05); 与模型组比 较,无促透剂组、氮酮组和冰片组Leptin、Leptin受体、JAK2和STAT3 mRNA表达显著升高(均P<0.05); 与无促透剂 组相比, 氮酮组和冰片组的Leptin、Leptin受体、JAK2和STAT3 mRNA表达显著升高(均P<0.05); 与氮酮组比较, 冰 片组的Leptin、Leptin受体、JAK2和STAT3 mRNA表达显著升高(均P<0.05)。免疫组化和WB检测趋势与qPCR结果基本一致。磷酸化JAK2 (Phospho-JAK2)和磷酸化STAT3 (phospho-STAT3)免疫组化和WB检测趋势与JAK2和STAT3基本一 致。

结论: 高脂模型兔的Leptin/JAK2/STAT3通路分子表达下降, 隔药饼灸后Leptin/JAK2/STAT3通路因子表达显著上 升, 氮酮和冰片作为促透剂运用于隔药饼灸比单纯隔药饼灸对Leptin/JAK2/STAT3调脂通路相关因子的上调作用更 明显。

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Acknowledgments

This work was supported by Hunan Province College Innovation Open-end Fund Project (湖南省高校创新平台开放基金项目, No. 16K068); 2017 Scientific and Research Innovative Project for Postgraduate Students of Hunan Province (2017 年湖南省研究生科研创新项目, No. CX2017B431).

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Authors and Affiliations

  1. Hunan University of Chinese Medicine, Changsha, 410208, China

    Zong-li Liao  (廖宗力), Jing Tan  (谭静), Lu Sun  (孙璐), Ren-da Yang  (阳仁达) & Xiao-rong Chang  (常小荣)

  2. Huizhou Hospital of Traditional Chinese Medicine, Guangdong, 516000, China

    Chong-zheng Zhu  (朱重政)

  3. The Second Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, 410005, China

    Wen-tao Huang  (黄文韬)

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  1. Zong-li Liao  (廖宗力)
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Correspondence to Xiao-rong Chang  (常小荣).

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Liao, Zl., Tan, J., Zhu, Cz. et al. Effect of herbal cake-partitioned moxibustion on Leptin/JAK2/STAT3 in lipid-lowering pathway of hyperlipidemia rabbits. J. Acupunct. Tuina. Sci. 17, 371–382 (2019). https://doi.org/10.1007/s11726-019-1139-2

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