1.Prediction of pulp exposure risk of carious pulpitis based on deep learning.
Li WANG ; Fei WU ; Mo XIAO ; Yu-Xin CHEN ; Ligeng WU
West China Journal of Stomatology 2023;41(2):218-224
OBJECTIVES:
This study aims to predict the risk of deep caries exposure in radiographic images based on the convolutional neural network model, compare the prediction results of the network model with those of senior dentists, evaluate the performance of the model for teaching and training stomatological students and young dentists, and assist dentists to clarify treatment plans and conduct good doctor-patient communication before surgery.
METHODS:
A total of 206 cases of pulpitis caused by deep caries were selected from the Department of Stomatological Hospital of Tianjin Medical University from 2019 to 2022. According to the inclusion and exclusion criteria, 104 cases of pulpitis were exposed during the decaying preparation period and 102 cases of pulpitis were not exposed. The 206 radiographic images collected were randomly divided into three groups according to the proportion: 126 radiographic images in the training set, 40 radiographic images in the validation set, and 40 radiographic images in the test set. Three convolutional neural networks, visual geometry group network (VGG), residual network (ResNet), and dense convolutional network (DenseNet) were selected to analyze the rules of the radiographic images in the training set. The radiographic images of the validation set were used to adjust the super parameters of the network. Finally, 40 radiographic images of the test set were used to evaluate the performance of the three network models. A senior dentist specializing in dental pulp was selected to predict whether the deep caries of 40 radiographic images in the test set were exposed. The gold standard is whether the pulp is exposed after decaying the prepared hole during the clinical operation. The prediction effect of the three network models (VGG, ResNet, and DenseNet) and the senior dentist on the pulp exposure of 40 radiographic images in the test set were compared using receiver operating characteristic (ROC) curve, area under the ROC curve (AUC), accuracy, sensitivity, specificity, positive predictive value, negative predictive value, and F1 score to select the best network model.
RESULTS:
The best network model was DenseNet model, with AUC of 0.97. The AUC values of the ResNet model, VGG model, and the senior dentist were 0.89, 0.78, and 0.87, respectively. Accuracy was not statistically different between the senior dentist (0.850) and the DenseNet model (0.850)(P>0.05). Kappa consistency test showed moderate reliability (Kappa=0.6>0.4, P<0.05).
CONCLUSIONS
Among the three convolutional neural network models, the DenseNet model has the best predictive effect on whether deep caries are exposed in imaging. The predictive effect of this model is equivalent to the level of senior dentists specializing in dental pulp.
Humans
;
Deep Learning
;
Neural Networks, Computer
;
Pulpitis/diagnostic imaging*
;
Reproducibility of Results
;
ROC Curve
;
Random Allocation
2.Technical recommendation for pragmatic randomized controlled trial of heat-sensitive moxibustion in community (Ⅰ): randomization with consideration of patient preference.
Xu ZHOU ; Ze-Huai WEN ; Ling LI ; Jian-Ping LIU ; Yi-Huang GU ; Xin-Feng GUO ; Xing LIAO ; Wei-Feng ZHU ; Shu-Qing LI ; Xin SUN
Chinese Acupuncture & Moxibustion 2022;42(1):85-90
Heat-sensitive moxibustion is the appropriate technique of the external treatment in traditional Chinese medicine and it is widely used in community because of its "easy learning, simple operation and clear curative effect". Pragmatic randomized controlled trial is a main intervention design in the real world study, which provides a high-level evidence for the effectiveness assessment of heat-sensitive moxibustion in community management. Focusing on the key links of randomization, e.g. block randomization, stratified randomization, cluster randomization, sample size allocation, allocation concealment and blinding, the paper elaborates the advantages, disadvantages and technical details of two-stage randomization with consideration of patient preference in pragmatic randomized controlled trials of heat-sensitive moxibustion in community. It facilitates improving the quality of evidence, reproducibility and methodological homogeneity among different trials.
Hot Temperature
;
Humans
;
Moxibustion
;
Patient Preference
;
Random Allocation
;
Reproducibility of Results
3.Xenon post-conditioning protects against spinal cord ischemia-reperfusion injury in rats by downregulating mTOR pathway and inhibiting endoplasmic reticulum stress-induced neuronal apoptosis.
Lan LUO ; Jia Qi TONG ; Lu LI ; Mu JIN
Journal of Southern Medical University 2022;42(8):1256-1262
OBJECTIVE:
The purpose of this study was to determine whether xenon post-conditioning affects mTOR signaling as well as endoplasmic reticulum stress (ERS)-apoptosis pathway in rats with spinal cord ischemia/reperfusion injury.
METHODS:
Fifty male rats were randomized equally into sham-operated group (Sham group), I/R model group (I/R group), I/R model+ xenon post-conditioning group (Xe group), I/R model+rapamycin (a mTOR signaling pathway inhibitor) treatment group (I/R+ Rapa group), and I/R model + xenon post- conditioning with rapamycin treatment group (Xe + Rapa group).. In the latter 4 groups, SCIRI was induced by clamping the abdominal aorta for 85 min followed by reperfusion for 4 h. Rapamycin (or vehicle) was administered by daily intraperitoneal injection (4 mg/kg) for 3 days before SCIRI, and xenon post-conditioning by inhalation of 1∶1 mixture of xenon and oxygen for 1 h at 1 h after initiation of reperfusion; the rats without xenon post-conditioning were given inhalation of nitrogen and oxygen (1∶ 1). After the reperfusion, motor function and histopathologic changes in the rats were examined. Western blotting and real-time PCR were used to detect the protein and mRNA expressions of GRP78, ATF6, IRE1α, PERK, mTOR, p-mTOR, Bax, Bcl-2 and caspase-3 in the spinal cord.
RESULTS:
The rats showed significantly lowered hind limb motor function following SCIRI (P < 0.01) with a decreased count of normal neurons, increased mRNA and protein expressions of GRP78, ATF6, IRE1α, PERK, and caspase-3, and elevated p-mTOR/mTOR ratio and Bax/Bcl-2 ratio (P < 0.01). Xenon post-conditioning significantly decreased the mRNA and protein levels of GRP78, ATF6, IRE1α, PERK and caspase-3 (P < 0.05 or 0.01) and reduced p-mTOR/mTOR and Bax/Bcl-2 ratios (P < 0.01) in rats with SCIRI; the mRNA contents and protein levels of GRP78 and ATF6 were significantly decreased in I/R+Rapa group (P < 0.01). Compared with those in Xe group, the rats in I/R+Rapa group and Xe+Rapa had significantly lowered BBB and Tarlov scores of the hind legs (P < 0.01), and caspase-3 protein level and Bax/Bcl-2 ratio were significantly lowered in Xe+Rapa group (P < 0.05 or 0.01).
CONCLUSION
By inhibiting ERS and neuronal apoptosis, xenon post- conditioning may have protective effects against SCIRI in rats. The mTOR signaling pathway is partially involved in this process.
Animals
;
Apoptosis
;
Caspase 3/metabolism*
;
Endoplasmic Reticulum Stress
;
Endoribonucleases/pharmacology*
;
Injections, Intraperitoneal
;
Male
;
Neurons/pathology*
;
Nitrogen/metabolism*
;
Oxygen/metabolism*
;
Protein Serine-Threonine Kinases
;
Proto-Oncogene Proteins c-bcl-2/metabolism*
;
RNA, Messenger/metabolism*
;
Random Allocation
;
Rats
;
Rats, Sprague-Dawley
;
Reperfusion Injury/metabolism*
;
Sirolimus/pharmacology*
;
Spinal Cord Ischemia/pathology*
;
TOR Serine-Threonine Kinases/metabolism*
;
Xenon/therapeutic use*
;
bcl-2-Associated X Protein/metabolism*
4.Effect of
Yi Qun LIU ; Ling He HUANG ; Pei Pei LIU ; Qing Bin XING ; Feng HAN ; Qin WANG ; Shu Rong CHEN ; Kimio SUGIYAMA ; Xue Song XIANG ; Zhen Wu HUANG
Biomedical and Environmental Sciences 2021;34(5):356-363
Objective:
This study aimed to investigate the effects of
Methods:
In this study, 0.1% DMG was supplemented in 20% casein diets that were either folate-sufficient (20C) or folate-deficient (20CFD). Blood and liver of rats were subjected to assays of Hcy and its metabolites. Hcy and its related metabolite concentrations were determined using a liquid chromatographic system.
Results:
Folate deprivation significantly increased pHcy concentration in rats fed 20C diet (from 14.19 ± 0.39 μmol/L to 28.49 ± 0.50 μmol/L;
Conclusion
DMG supplementation exhibited hypohomocysteinemic effects under folate-sufficient conditions. By contrast, the combination of folate deficiency and DMG supplementation has deleterious effect on pHcy concentration.
Animals
;
Biomarkers/metabolism*
;
Chromatography, Liquid
;
Diet
;
Dietary Supplements
;
Folic Acid Deficiency/metabolism*
;
Homocysteine/metabolism*
;
Liver/metabolism*
;
Male
;
Random Allocation
;
Rats
;
Rats, Wistar
;
Sarcosine/metabolism*
5.The mechanism of enriched environment repairing the learning and memory impairment in offspring of prenatal stress by regulating the expression of activity-regulated cytoskeletal-associated and insulin-like growth factor-2 in hippocampus.
Su-Zhen GUAN ; You-Juan FU ; Feng ZHAO ; Hong-Ya LIU ; Xiao-Hui CHEN ; Fa-Qiu QI ; Zhi-Hong LIU ; Tzi Bun NG
Environmental Health and Preventive Medicine 2021;26(1):8-8
BACKGROUND:
Prenatal stress can cause neurobiological and behavioral defects in offspring; environmental factors play a crucial role in regulating the development of brain and behavioral; this study was designed to test and verify whether an enriched environment can repair learning and memory impairment in offspring rats induced by prenatal stress and to explore its mechanism involving the expression of insulin-like growth factor-2 (IGF-2) and activity-regulated cytoskeletal-associated protein (Arc) in the hippocampus of the offspring.
METHODS:
Rats were selected to establish a chronic unpredictable mild stress (CUMS) model during pregnancy. Offspring were weaned on 21st day and housed under either standard or an enriched environment. The learning and memory ability were tested using Morris water maze and Y-maze. The expression of IGF-2 and Arc mRNA and protein were respectively measured by using RT-PCR and Western blotting.
RESULTS:
There was an elevation in the plasma corticosterone level of rat model of maternal chronic stress during pregnancy. Maternal stress's offspring exposed to an enriched environment could decrease their plasma corticosterone level and improve their weight. The offspring of maternal stress during pregnancy exhibited abnormalities in Morris water maze and Y-maze, which were improved in an enriched environment. The expression of IGF-2, Arc mRNA, and protein in offspring of maternal stress during pregnancy was boosted and some relationships existed between these parameters after being exposed enriched environment.
CONCLUSIONS
The learning and memory impairment in offspring of prenatal stress can be rectified by the enriched environment, the mechanism of which is related to the decreasing plasma corticosterone and increasing hippocampal IGF-2 and Arc of offspring rats following maternal chronic stress during pregnancy.
Animals
;
Cytoskeletal Proteins/metabolism*
;
Female
;
Gene Expression Regulation
;
Hippocampus/metabolism*
;
Insulin-Like Growth Factor II/metabolism*
;
Learning
;
Learning Disabilities/psychology*
;
Male
;
Memory Disorders/psychology*
;
Nerve Tissue Proteins/metabolism*
;
Pregnancy
;
Prenatal Exposure Delayed Effects/psychology*
;
Random Allocation
;
Rats
;
Rats, Wistar
;
Social Environment
;
Stress, Psychological/genetics*
6.Effect of acupuncture on intestinal flora in rats with stress gastric ulcer.
Liu-Jing WANG ; Ting XUE ; Ying-Qi WU ; Jia-Yu ZHAO ; Tu-Nan WANG ; Jing-Ting LI ; Chen-Lu FU ; Jia-Jia MA ; Li-Ping ZHANG ; Yi-Xuan SHAO ; Yi-Chen YANG ; Zi-Xian ZHOU ; Hui-Fang MA
Chinese Acupuncture & Moxibustion 2020;40(5):526-532
OBJECTIVE:
To observe the effect of acupuncture at "Baihui" (GV 20), "Zhongwan" (CV 12) and "Zusanli" (ST 36) on intestinal flora in rats with stress gastric ulcer (SGU) , and to explore the mechanism of acupuncture promoting SGU recovery.
METHODS:
Thirty-one SPF SD rats were randomly divided into a control group (7 rats), a model control group (8 rats), an acupuncture group (8 rats) and a medication group (8 rats). The rats in the model group, acupuncture group and medication group were selected to applied the improved restraint water-immersion stress method to establish the SGU model. After modeling, the rats in the control group and model group were fixed and restrained for 20 min every day for a total of 5 days; the rats in the acupuncture group were intervented with acupuncture at "Baihui" (GV 20), "Zhongwan" (CV 12) and "Zusanli" (ST 36), once a day, 20 min each time, and twisting needle for 30 s every 5 min for a total of 5 days; the rats in the medication group were gavaged by solution of omeprazole enteric-coated tablet (200 mg/mL), 2 mL for each rat, once a day. Guth method was used to calculate the gastric mucosal damage index (GMDI), HE staining was used to observe the pathological changes of gastric mucosa, and 16SrDNA identification was used to detect the structural abundance of intestinal flora.
RESULTS:
Compared with the control group, the GMDI of rats in the model group was increased (<0.01), the gastric mucosal pathological changes were significant, and the intestinal flora richness index Chao1, Observed species and diversity index Shannon were all decreased (<0.05), the diversity index Simpson was increased (<0.05). Compared with the model group, the GMDI of rats in the acupuncture group and medication group was reduced (<0.01, <0.05), the gastric mucosal damage degree was reduced, and the intestinal flora richness index Chao1, Observed species and diversity index Shannon were all increased (<0.05) and the diversity index Simpson decreased (<0.05). Compared with the medication group, the GMDI of rats in the acupuncture group was reduced (<0.01), the recovery of gastric mucosal injury was better than that of the medication group.
CONCLUSION
Acupuncture can effectively improve gastric mucosal injury of SGU, and the mechanism may be related to increasing the diversity of intestinal flora and promoting the correction of the disordered intestinal flora.
Acupuncture Points
;
Acupuncture Therapy
;
Animals
;
Gastrointestinal Microbiome
;
Random Allocation
;
Rats
;
Rats, Sprague-Dawley
;
Stomach Ulcer
;
microbiology
;
therapy
7.Effect of electroacupuncture at "Jiaji" (EX-B 2) points on the proliferation and differentiation of oligodendrocyte precursor cells in rats with acute spinal cord injury.
Rong HU ; Yi CHEN ; Hai-Peng XU ; Ke-Lin HE ; Lian-Zhu SUN ; Lei WU ; Rui-Jie MA
Chinese Acupuncture & Moxibustion 2020;40(5):519-525
OBJECTIVE:
To observe the effect of electroacupuncture (EA) at "Jiaji" (EX-B 2) points on the proliferation and differentiation of oligodendrocyte precursor cells in rats with acute incomplete spinal cord injury, and to explore the mechanism of EA on improving motor function of spinal cord injury.
METHODS:
A total of 72 male SPF SD rats were randomly divided into a sham operation group, a model group, an EA group and a medication group, 18 rats in each group. Each group was further divided into 1-day subgroup, 7-day subgroup and 14-day subgroup, 6 rats in each subgroup. The T acute incomplete spinal cord injury model was established by modified Allen's method in the model group, EA group and medication group. The rats in each group received intraperitoneal injection of 5-bromodeoxyuridine (BrdU, 50 mg/kg), once a day, and each subgroup received continuous injection for 1, 7, 14 times for cell proliferation labeling. The rats in the EA group were treated with EA at "Jiaji" (EX-B 2) points 3-4 mm next the spinous process of the upper and lower segments of the injured spinal cord (T, T) with a frequency of 2 Hz/100 Hz and intensity of 1-2 mA. The muscle twitch at the treatment site was taken as the degree. The treatment was given 20 min each time, once a day. In the medication group, monosialogangliosides (GM1) was injected intraperitoneally (10 mg/kg), once a day. The subgroups of EA group and medication group were treated for 1, 7, 14 times. The score of Basso Beattie Bresnahan (BBB) was used to evaluate the motor function of hind limbs. The co-expression of BrdU/NG2 positive cells was detected by immunofluorescence, and the expression of Olig2 and Sox10 was detected by Western blot.
RESULTS:
Compared with the sham operation group, the BBB score was decreased 1 day, 7 days and 14 days after operation in the model group (<0.05), the expression of Olig2 and Sox10 was increased (<0.05), and the co-expression of BrdU/NG2 positive cells was increased 7 days and 14 days after operation (<0.05). Seven days and 14 days after operation, the BBB score in the EA group and medication group was higher than that in the model group (<0.05), and the co-expression of BrdU/NG2 in the medication group was higher than that in the model group (<0.05). Fourteen days after operation, the co-expression of BrdU/NG2 in the EA group was higher than that in the model group (<0.05); 1 day, 7 days and 14 days after operation, the expression of Olig2 and Sox10 in the EA group and medication group was higher than that in the model group (<0.05). Compared with the medication group, the co-expression of BrdU/NG2 positive cells in the EA group 14 days after operation was decreased (<0.05); 1 day, 7 days and 14 days after operation, the expression of Olig2 and Sox10 in the EA group was decreased (<0.05).
CONCLUSION
EA at "Jiaji" (EX-B 2) points could promote the expression of Olig2 and Sox10 after spinal cord injury, which has similar effects with GM1. It could promote the proliferation and differentiation of oligodendrocyte precursor cells into oligodendrocytes, so as to promote the recovery of motor function of rats.
Acupuncture Points
;
Animals
;
Cell Differentiation
;
Cell Proliferation
;
Electroacupuncture
;
Humans
;
Male
;
Oligodendrocyte Precursor Cells
;
cytology
;
Oligodendrocyte Transcription Factor 2
;
metabolism
;
Random Allocation
;
Rats
;
Rats, Sprague-Dawley
;
SOXE Transcription Factors
;
metabolism
;
Spinal Cord
;
Spinal Cord Injuries
;
therapy
8.Effect of electroacupuncture on the expression of insulin signal pathway related proteins in hippocampus in mice with Alzheimer's disease.
Yan-Jun WU ; Kai-Hui WU ; Rong JIANG ; Yi-Ying WANG ; Min LUO ; Qing-Qian XIA ; Cheng-Lin TANG ; Shu-Juan ZHU ; Hua-Jun SHENG
Chinese Acupuncture & Moxibustion 2020;40(5):513-518
OBJECTIVE:
To observe the effect of electroacupuncture (EA) on the expression of insulin phosphatidylinositol-3 kinase/glycogen synthetase kinase-3α (PI3K/GSK3α) signal pathway related proteins in the hippocampus in mice with Alzheimer's disease (AD), and to explore the regulatory mechanism of EA on improving the pathological characteristics of AD.
METHODS:
Twelve male APP/PS1 double transgenic mice were randomly divided a model group and a treatment group, 6 mice in each group; another 6 wild-type male mice were taken as the control group. The mice in the treatment group were treated with EA (continuous wave, 2 Hz of frequency) at "Baihui" (GV 20) and bilateral "Shenshu" (BL 23), once a day; 7-day treatment was taken as a course of treatment, and 2 courses of treatment were given. The immunohistochemistry method and Western blot method were used to detect the distribution and expression level of hippocampal PI3K/GSK3α signal pathway related proteins P85α, P110α, GSK3α and pSGSK3α, and the number of hippocampal senile plaques (SP) was observed.
RESULTS:
The proteins of P85α, P110α, GSK3α and pSGSK3α were mainly distributed in the cytoplasm of hippocampal neurons, and the GSK3α was also distributed in the axons of neurons in the model group and the treatment group. The immunohistochemistry results showed that the distribution level of GSK3α in the hippocampus in the model group was significantly higher than that in the control group (<0.001), and the distribution level of pSGSK3α, P85α and P110α was significantly decreased (<0.01, <0.001); compared with the model group, the distribution level of GSK3α in the hippocampus in the treatment group was significantly decreased (<0.001), and the distribution level of pSGSK3α, P85α and P110α in hippocampus was significantly increased (<0.05, <0.001). The Western blot results showed compared with the control group, the expression of pSGSK3α, P85α and P110α as well as the ratio of pSGSK3α/GSK3α in the hippocampus in the model group were significantly decreased (<0.001), and the expression of GSK3α was increased (<0.05); compared with the model group, the expression of pSGSK3α, P85α, P110α and the ratio of pSGSK3α/GSK3α in the hippocampus in the treatment group were significantly increased (<0.01, <0.001), and the expression of GSK3α was decreased (<0.05). Compared with the control group, the number of hippocampal SP in the model group was significantly increased (<0.001); compared with the model group, the number of hippocampal SP in the treatment group was significantly decreased (<0.01).
CONCLUSION
EA could effectively regulate the expression of PI3K/GSK3α signal pathway related proteins in the hippocampus in mice with AD, so as to reduce the formation and deposition of SP.
Alzheimer Disease
;
therapy
;
Animals
;
Electroacupuncture
;
Hippocampus
;
physiology
;
Insulin
;
physiology
;
Male
;
Mice
;
Mice, Transgenic
;
Random Allocation
;
Signal Transduction
9.Early intervention of bone-nearby acupuncture combined with electroacupuncture on morphine tolerance in bone cancer pain rats and its effect on the expression of HDAC and MOR in dorsal root ganglia.
Xue-Mei ZHONG ; Jun-Fan FANG ; Bin JIANG ; Jie ZHOU ; Yang-Qian CAI ; Yi LIANG ; Jian-Qiao FANG ; Feng CHEN ; Jun-Ying DU
Chinese Acupuncture & Moxibustion 2020;40(4):405-410
OBJECTIVE:
To observe the effect of early intervention of bone-nearby acupuncture (BNA) combined with electroacupuncture (EA) on the expression of histone deacetylase1(HDAC1), histone deacetylase 2 (HDAC2) andμ-opioid recepter (MOR) in dorsal root ganglia (DRG) of bone cancer pain-morphine tolerance (BCP-MT) rats, and to explore its possible mechanism.
METHODS:
A total of 35 SD rats were randomized into a sham BCP group (=6), a BCP group (=7), a MT group (=7), a BNA+EA group (=8) and a shame BNA group (=7). Except of the sham BCP group, cancer cell inoculation operation at left tibia was given in the other 4 groups to establish the bone cancer pain model. In the MT group, the BNA+EA group and the shame BNA group, intraperitoneal injection of morphine hydrochloride was given to establish the morphine tolerance model. After the operation, bone-nearby acupuncture combined with electroacupuncture was applied at "Zusanli" (ST 36) and "Kunlun" (BL 60) in the BNA+EA group, with dilatational wave, 2 Hz/100 Hz in frequency, 0.5 to 1.5 mA in intensity. Intervention in the shame BNA group was applied at the same time and acupoints as those in the BNA+EA group, the needles were pierced the skin without any electrical stimulation. The needles were retained for 30 min, once a day for continuous 7 days in both BNA+EA and shame BNA groups. Before and 10, 11, 15, 22 days after the operation, the left paw withdrawal threshold (PWT) was measured in the 5 groups. The levels of HDAC1, HDAC2 and MOR in DRG were detected by Western blot.
RESULTS:
Ten days after the cancer cell inoculation operation, the PWT of the BCP, MT, BNA+EA and sham BNA groups was decreased compared with the sham BCP group (<0.01). Eleven days after the operation, the PWT of the MT, BNA+EA and sham BNA groups was increased compared with the BCP group (<0.01). Twenty-two days after the operation, the difference was no significant between the BCP group and MT group (>0.05); the PWT of the BNA+EA group was increased compared with the MT and sham BNA group (<0.01). In the BCP group, the DRG levels of HDAC1 and HDCA2 were increased, while the level of MOR was decreased compared with the sham BCP group (<0.05, <0.01). In the MT group, the DRG level of HDAC1 was increased compared with the BCP group (<0.05). In the BNA+EA group, the DRG level of HDAC1 was decreased compared with the MT group and the sham BNA group (<0.01, <0.05), while the level of MOR was increased (<0.01).
CONCLUSION
Early intervention of bone-nearby acupuncture combined with electroacupuncture can relieve the morphine tolerance in bone cancer pain rats, it may relate to down-regulating the expression of HDAC1 and up-regulating the expression of MOR in the dorsal root ganglia.
Acupuncture Points
;
Animals
;
Bone Neoplasms
;
complications
;
Cancer Pain
;
therapy
;
Drug Tolerance
;
Electroacupuncture
;
Ganglia, Spinal
;
metabolism
;
Histone Deacetylases
;
metabolism
;
Morphine
;
Random Allocation
;
Rats
;
Rats, Sprague-Dawley
;
Receptors, Opioid, mu
;
metabolism
10.Comparison of the effect between electroacupuncture and NSAIDs on pain memory based on cAMP/PKA/CREB pathway in anterior cingulate gyrus.
Jing SUN ; Jian-Qiao FANG ; Zui SHEN ; Yi-Lin ZHU ; Qin CHEN ; Fang FANG ; Jia-Ling WANG ; Fei LI ; Xiao-Mei SHAO
Chinese Acupuncture & Moxibustion 2020;40(4):397-404
OBJECTIVE:
To observe the direct intervention effects of electroacupuncture (EA) and non-steroid anti-inflammatory drugs (NSAIDs) on pain memory, and to explore their effects on cAMP/PKA/cAMP pathway in anterior cingulate gyrus (ACC).
METHODS:
Fifty clean healthy male SD rats were randomly divided into a control group, a model group, an indomethacin group, an EA group and a sham EA group, 10 rats in each group. Except the control group, the pain memory model was established in the remaining four groups by twice injection of carrageenan at foot; 0.1 mL of 2%λ-carrageenan was subcutaneously injected at the left foot of rats; 14 days later, when the pain threshold of rats of each group returned to the basic level, the second injection was performed with the same procedure. The rats in the EA group were treated with EA at bilateral "Zusanli" (ST 36) for 30 min; the rats in the indomethacin group was treated with indomethacin intragastric administration with the dose of 3 mg/kg; the rats in the sham EA group was treated with EA without electricity at the point 0.3 mm forward "Zusanli" (ST 36) with the depth of 2 mm for 30 min; the rats in the control group was not given any invention. All the above interventions were performed 5 h, 1 d, 2 d and 3 d after the second injection of 2% λ-carrageenan. The left-side paw withdrawal thresholds (PWT) were observed before the first injection, 4 h, 3 d, 5 d after the first injection, before the second injection and 4 h, 1 d, 2 d, 3 d after the second injection. Three days after the second injection, the number of positive cells of cAMP, p-PKA, p-CREB and the number of positive cells of protein co-expression in the right ACC brain area were detected by immunofluorescence, and the relative protein expression of p-PKA and p-CREB were detected by Western blot.
RESULTS:
Compared with the control group, the PWTs in the model group decreased significantly 4 h, 3 d and 5 d after the first injection and 1 d, 2 d and 3 d after the second injection (<0.05); compared with the control group, the positive expression of cAMP, p-PKA and p-CREB in the right ACC brain area in the model group increased significantly (<0.05), and the number of positive cells of the co-expression of cAMP/p-PKA and p-PKA/p-CREB also increased significantly (<0.05). Compared with the model group, indomethacin group and sham EA group, the PWTs in the EA group were increased significantly 1 d, 2 d and 3 d after the second injection (<0.05); compared with the model group, indomethacin group and sham EA group, the positive expression of p-PKA and p-CREB in the right ACC brain area in the EA group decreased significantly (<0.05), and the number of positive cells of co-expression of cAMP/p-PKA and p-PKA/p-CREB was decreased significantly (<0.05). Compared with the model group and sham EA group, the positive expression of cAMP in the right ACC brain area was decreased in the EA group (<0.05).
CONCLUSION
EA have a direct intervention effect on pain memory, which have significant advantage over NSAIDs in the treatment of chronic pain. The advantage effect of EA on pain memory may be related to the inhibition of cAMP/PKA/CREB pathway in ACC area.
Animals
;
Anti-Inflammatory Agents, Non-Steroidal
;
therapeutic use
;
Cyclic AMP
;
metabolism
;
Cyclic AMP Response Element-Binding Protein
;
metabolism
;
Cyclic AMP-Dependent Protein Kinases
;
metabolism
;
Electroacupuncture
;
Gyrus Cinguli
;
metabolism
;
Male
;
Pain Threshold
;
Random Allocation
;
Rats
;
Rats, Sprague-Dawley
;
Signal Transduction

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