Oxford Nanopore Technologies (ONT) Full-length Transcriptomics Shows Electroacupuncture ameliorates Lipid Metabolic Disorder through Suppressing Hepatic Pdia3/Perk/Qrich1 Expression in Rats
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: June 2024

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.

Objectives:
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.