ONT Full-length Transcriptomics Shows Electroacupuncture Improves Lipid Metabolic Disorder through Pdia3/Perk/Qrich1 Signal Pathway
Authors: Zhang Pu , Li Yue , Zhang Ning , Wang Xiao-gang , Qu Yan-wei , Pei Hui , Liu Xiao-chun , Liu Qi , Lv Pei-ran , Zhao Xian 
Published: December 2022

Abstract
Background: The incidence of dyslipidemia increases after menopause. Electroacupuncture (EA) has some advantages and potential for the development of treatment of menopause-related disorders. But, the positive effect on lipid metabolism disorders remains unclear.
Objectives: To investigate the underline mechanism of EA treatment on the lipid metabolism disorders through ONT full‑length transcriptome sequencing.
Methods: Adult Female SD rats were randomly divided into Ctrl, Sham+HFD, OVX+HFD, OVX+HFD+ATO and OVX+HFD+EA groups. Adipose tissue around bilateral ovaries of rats in 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 OVX+HFD+EA group. ST36, PC6, SP6, BL18 and ST40 were the selected acupoints. Track daily food intake. Body weights were recorded weekly. The samples were collected after 30 days of treatment. The serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein (HDL-C) were detected. 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, RT‑qPCR and immunofluorescence staining were used to verify the crucial signal pathway screened by the ONT full‑length transcriptome sequencing.
Results:The results showed that the weight of perirenal fat and liver was decreased after EA treatment. The volume of liver tissue in the OVX+HFD group became larger, and the liver color showed light yellow, showing that it may have fatty degeneration. After EA and ATO treatment, it had obvious improvement. Also, EA could improve the lipid profile and hepatic steatosis in OVX+HFD rats. Ground on the full‑length transcriptome sequencing, 2292 genes showing differential expressed in the OVX+HFD group; of these, 1121 up-regulated 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 obtained 77 genes significantly up-regulated after EA intervention through Veen map analysis (including Agtr1a, Pdia3, etc.), which may be the goal genes for EA treatment lipid metabolism disorders. Finally, we verified the expression of Pdia3, Perk and Qrich1 level 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. Pdia3/Perk/Qrich1 signal pathway may play crucial roles in the improvement of lipid metabolism disorder of OVX+HFD rats after EA treatment.