Abstract：
BACKGROUND: The incidence of dyslipidemia increases after menopause. Electroacupuncture (EA) has been recommended for menopause-related disease. However, the positive effect on lipid metabolism disorders is still unclear.
OBJECTIVE: To investigate the underlying mechanism of EA treatment on lipid metabolism disorders through ONT full-length transcriptome sequencing Methods: Adult female SD rats were randomly divided into Ctrl, sham operation+high-fat feed(Sham+HFD), Ovariectomized+high-fat feed (OVX+HFD), Ovariectomized+high-fat feed + Atorvastatin (OVX+HFD+ATO) and OVX+HFD+EA groups. Periovarian adipose tissue around the bilateral ovaries of rats in the Sham+HFD group was resected. Rats in the OVX+HFD, OVX+HFD+ATO and OVX+HFD+EA groups were subjected to bilateral oophorectomy to prepare the ovariectomized rat model. Treatment was applied to rats in the OVX+HFD+EA group. ST36, PC6, SP6, BL18 and ST40 were the selected acupoints. Daily food intake and body weights of rats were recorded. The samples were collected 30 days after treatment. The serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein (HDL-C) were detected to assess the improvement of lipid metabolism disorders. HE and oil red O staining were used to stain the liver tissues. Total RNA was extracted from liver tissues, and its transcriptional changes were determined by high-throughput sequencing. Additionally, RTÁqPCR and immunofluorescence staining were used to verify the crucial signal pathway screened by the ONT fullÁlength transcriptome sequencing.
RESULTS: EA treatment resulted in a lowered weight of perirenal fat and liver and a significant improvement in the color of the liver. In addition, EA could improve the lipid profile and hepatic steatosis in OVX+HFD rats. According to fullÁlength transcriptome sequencing, 2292 genes showed differential expression in the OVX+HFD group; of these, 1121 were upregulated and 1171 down-regulated. 609 DEGs were found in the OVX+HFD+EA group compared to the OVX+HFD group; 235 up-regulated and 374 down-regulated. We also found that 77 genes are significantly upregulated after EA intervention through Venn map analysis (including Agtr1a, Pdia3, etc.), which may be the targeted genes for EA treatment of lipid metabolism disorders. Finally, we verified the expression of Pdia3, Perk and Qrich1 levels in liver tissues. HFD feeding could increase the expression of Pdia3 and its downstream signal pathways molecular Perk and Qrich1. But these effects were reversed by EA treatment, the results demonstrated that the expression of pdia3, Perk, as well as Qrich1 of OVX+HFD rats had a decreasing trend after EA treatment.
CONCLUSIONS: EA could ameliorate lipid metabolic disorder in OVX+HFD rats. The Pdia3/Perk/Qrich1 signal pathway may play crucial roles in the improvement of lipid metabolism disorder of OVX+HFD rats after EA treatment.
OBJECTIVE: To investigate the underlying mechanism of EA treatment on lipid metabolism disorders through ONT full-length transcriptome sequencing Methods: Adult female SD rats were randomly divided into Ctrl, sham operation+high-fat feed(Sham+HFD), Ovariectomized+high-fat feed (OVX+HFD), Ovariectomized+high-fat feed + Atorvastatin (OVX+HFD+ATO) and OVX+HFD+EA groups. Periovarian adipose tissue around the bilateral ovaries of rats in the Sham+HFD group was resected. Rats in the OVX+HFD, OVX+HFD+ATO and OVX+HFD+EA groups were subjected to bilateral oophorectomy to prepare the ovariectomized rat model. Treatment was applied to rats in the OVX+HFD+EA group. ST36, PC6, SP6, BL18 and ST40 were the selected acupoints. Daily food intake and body weights of rats were recorded. The samples were collected 30 days after treatment. The serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein (HDL-C) were detected to assess the improvement of lipid metabolism disorders. HE and oil red O staining were used to stain the liver tissues. Total RNA was extracted from liver tissues, and its transcriptional changes were determined by high-throughput sequencing. Additionally, RTÁqPCR and immunofluorescence staining were used to verify the crucial signal pathway screened by the ONT fullÁlength transcriptome sequencing.
RESULTS: EA treatment resulted in a lowered weight of perirenal fat and liver and a significant improvement in the color of the liver. In addition, EA could improve the lipid profile and hepatic steatosis in OVX+HFD rats. According to fullÁlength transcriptome sequencing, 2292 genes showed differential expression in the OVX+HFD group; of these, 1121 were upregulated and 1171 down-regulated. 609 DEGs were found in the OVX+HFD+EA group compared to the OVX+HFD group; 235 up-regulated and 374 down-regulated. We also found that 77 genes are significantly upregulated after EA intervention through Venn map analysis (including Agtr1a, Pdia3, etc.), which may be the targeted genes for EA treatment of lipid metabolism disorders. Finally, we verified the expression of Pdia3, Perk and Qrich1 levels in liver tissues. HFD feeding could increase the expression of Pdia3 and its downstream signal pathways molecular Perk and Qrich1. But these effects were reversed by EA treatment, the results demonstrated that the expression of pdia3, Perk, as well as Qrich1 of OVX+HFD rats had a decreasing trend after EA treatment.
CONCLUSIONS: EA could ameliorate lipid metabolic disorder in OVX+HFD rats. The Pdia3/Perk/Qrich1 signal pathway may play crucial roles in the improvement of lipid metabolism disorder of OVX+HFD rats after EA treatment.
摘要：
背景：绝经后血脂异常的发生率增加。电针（EA）已被推荐用于更年期相关疾病。然而,对脂质代谢紊乱的积极作用尚不清楚.
目的：通过ONT全长转录组测序研究EA治疗脂质代谢紊乱的潜在机制方法：成年雌性SD大鼠随机分为Ctrl,假手术+高脂饲料（Sham+HFD），去卵巢+高脂饲料（OVX+HFD），卵巢切除+高脂饲料+阿托伐他汀(OVX+HFD+ATO)和OVX+HFD+EA组。切除Sham+HFD组大鼠双侧卵巢周围的卵巢周围脂肪组织。OVX+HFD中的大鼠，OVX+HFD+ATO和OVX+HFD+EA组行双侧卵巢切除术制备去卵巢大鼠模型。对OVX+HFD+EA组的大鼠进行治疗。选择的穴位为ST36，PC6，SP6，BL18和ST40。记录大鼠的每日食物摄入量和体重。在处理后30天收集样品。血清总胆固醇(TC)水平,甘油三酯（TG），检测低密度脂蛋白胆固醇（LDL-C）和高密度脂蛋白（HDL-C）以评估脂质代谢紊乱的改善。HE和油红O染色用于肝脏组织染色。从肝脏组织中提取总RNA，并通过高通量测序确定其转录变化。此外,RTÁqPCR和免疫荧光染色用于验证ONTfullÁ长度转录组测序筛选的关键信号通路。
结果：EA治疗导致肾周脂肪和肝脏重量降低，肝脏颜色明显改善。此外,EA可以改善OVXHFD大鼠的血脂和肝脏脂肪变性。根据完整的Á长度转录组测序，2292个基因在OVX+HFD组中表现出差异表达；其中，1121被上调,1171被下调。与OVX+HFD组相比,在OVX+HFD+EA组中发现609个DEGs;235个上调和374个下调。我们还通过Venn图谱分析发现，EA干预后77个基因显著上调（包括Agtr1a，Pdia3等.),这可能是EA治疗脂质代谢紊乱的靶向基因。最后,我们验证了Pdia3，Perk和Qrich1在肝组织中的表达水平。HFD饲喂可以增加Pdia3及其下游信号通路分子Perk和Qrich1的表达。但是这些效果被EA治疗逆转了，结果表明，pdia3，Perk的表达，经EA处理后，OVX+HFD大鼠的Qrich1也有下降趋势。
结论：EA可以改善OVX+HFD大鼠的脂质代谢紊乱。Pdia3/Perk/Qrich1信号通路可能在改善OVX+HFD大鼠电针治疗后脂代谢紊乱中起重要作用。
目的：通过ONT全长转录组测序研究EA治疗脂质代谢紊乱的潜在机制方法：成年雌性SD大鼠随机分为Ctrl,假手术+高脂饲料（Sham+HFD），去卵巢+高脂饲料（OVX+HFD），卵巢切除+高脂饲料+阿托伐他汀(OVX+HFD+ATO)和OVX+HFD+EA组。切除Sham+HFD组大鼠双侧卵巢周围的卵巢周围脂肪组织。OVX+HFD中的大鼠，OVX+HFD+ATO和OVX+HFD+EA组行双侧卵巢切除术制备去卵巢大鼠模型。对OVX+HFD+EA组的大鼠进行治疗。选择的穴位为ST36，PC6，SP6，BL18和ST40。记录大鼠的每日食物摄入量和体重。在处理后30天收集样品。血清总胆固醇(TC)水平,甘油三酯（TG），检测低密度脂蛋白胆固醇（LDL-C）和高密度脂蛋白（HDL-C）以评估脂质代谢紊乱的改善。HE和油红O染色用于肝脏组织染色。从肝脏组织中提取总RNA，并通过高通量测序确定其转录变化。此外,RTÁqPCR和免疫荧光染色用于验证ONTfullÁ长度转录组测序筛选的关键信号通路。
结果：EA治疗导致肾周脂肪和肝脏重量降低，肝脏颜色明显改善。此外,EA可以改善OVXHFD大鼠的血脂和肝脏脂肪变性。根据完整的Á长度转录组测序，2292个基因在OVX+HFD组中表现出差异表达；其中，1121被上调,1171被下调。与OVX+HFD组相比,在OVX+HFD+EA组中发现609个DEGs;235个上调和374个下调。我们还通过Venn图谱分析发现，EA干预后77个基因显著上调（包括Agtr1a，Pdia3等.),这可能是EA治疗脂质代谢紊乱的靶向基因。最后,我们验证了Pdia3，Perk和Qrich1在肝组织中的表达水平。HFD饲喂可以增加Pdia3及其下游信号通路分子Perk和Qrich1的表达。但是这些效果被EA治疗逆转了，结果表明，pdia3，Perk的表达，经EA处理后，OVX+HFD大鼠的Qrich1也有下降趋势。
结论：EA可以改善OVX+HFD大鼠的脂质代谢紊乱。Pdia3/Perk/Qrich1信号通路可能在改善OVX+HFD大鼠电针治疗后脂代谢紊乱中起重要作用。
