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神経科学

Enhancing the Development and Growth of Infant Cerebral Palsy Rats Using Selective Spinal Manipulations

Published: February 02, 2024
doi:
10.3791/65659
, , , , , , , ,
1Yunnan University of Traditional Chinese Medicine

概要

This study illustrates the effect of selective spinal manipulation on the growth and development of infant rats with cerebral palsy, emphasizing the specific procedure and standardized protocol. Body weight measurement, Rotarod test, Foot-fault score, other behavioral tests, and growth hormone detection were performed to evaluate the protocol.

Abstract

Cerebral palsy (CP) is a refractory pediatric disease with a high prevalence, high disability rate, and difficult treatment. A variety of treatments are currently used for CP. The treatment involves drug and non-drug therapy. Traditional Chinese medicine external therapy is a very distinctive treatment method in non-drug therapy. As one of the external therapies of traditional Chinese medicine, massage is used in treating cerebral palsy and has good efficacy, small side effects, and strong operability. As a part of TCM external therapy, selective spinal manipulation can effectively promote the growth and development of infant rats with cerebral palsy.The operation was mainly divided into four steps: first, the rubbing method was applied to the spine and both sides of the spine for 1 min. The pressing and kneading method was applied to the spine for 5 min, and the muscles on both sides of the spine for 5 min. Second, pressing and kneading the sensitive local acupoints in the spine for 2 min were performed. Thirdly, the affected limb was treated by twisting method for 1 min. Fourth, the rubbing method was applied to a midline from the forehead to the back of the brain for 1 min. This study aimed to use selective spinal manipulation to treat infant rats with cerebral palsy. The weight, Rotarod test, Foot-fault score, and growth hormone of infant rats with cerebral palsy were detected to understand the effect of selective spinal manipulation on the growth and development of infant rats with cerebral palsy. The results showed that it can promote weight gain, improve balance ability and motor function, promote growth and development of infant cerebral palsy rats, promote growth hormone secretion, and increase the temperature of sensitive parts of the back.

Introduction

Cerebral Palsy (CP), caused by non-progressive damage to the brain in the fetus or infancy, is a group of disorders characterized by abnormal motor and postural development1, developmental disorders including slow weight gain2,3,4, and motor dysfunction. The incidence of cerebral palsy in China is 2.48%, and the prevalence is 2.46‰ (1-6 years old)5, respectively. Foreign studies reported a prevalence of 2.4%-3.6%6. Cerebral palsy is a major cause of physical injury and disability in children, among which balance and movement disorders significantly impact daily living activities1. Cerebral palsy can be caused by premature birth, infection, genetics, neonatal ischemia and hypoxia, neonatal jaundice, and other complex and diverse pathogenic factors7. The goal of treatment for cerebral palsy is to improve physical function and quality of life 8. Currently, the treatment methods for cerebral palsy include deep brain stimulation9, robot-assisted gait training10, wrist and ankle acupuncture11, and meridian acupuncture combined with massage12.

Traditional Chinese medicine (TCM) has been increasingly used as an effective treatment for CP13,14. Massage, as a part of it, also plays a certain role. For example, selective spinal manipulation can regulate the state of DNA hydroxymethylation to regulate neural development and improve learning and memory function15. In the treatment of infant rat with cerebral palsy, selective spinal manipulation may improve the inflammatory homeostasis of the cortex and hippocampus by regulating the methylation level of inflammatory cytokines TNF-α and IL-10 gene promoter region16 and play a positive therapeutic role in the balance ability of infant rat with cerebral palsy16. Selective spinal manipulation can improve the cognitive function of infant rats with cerebral palsy17. Selective spinal manipulation was used in the treatment of infant rats with cerebral palsy, and it was observed that it also had therapeutic effects on their growth and development18.

The purpose of this study is to illustrate the effect of selective spinal manipulation on the growth and development of infant rats with cerebral palsy by measuring the body weight, Rotarod test19, Foot-fault score20,21, and other behavioral tests and the detection of growth hormone, and to provide research ideas for relevant personnel.

Protocol

This study was approved by the Experimental Animal Ethics Committee of Yunnan University of Traditional Chinese Medicine. All experimental operations on animals followed the 3R principle of experimental animal reduction, optimization, and replacement (No. R-06202018). Healthy Sprague Dawley (SD) rats (14 males and 7 females) of Specific pathogen-free (SPF) grade with an average body weight of 250-300 g were used in this experiment. The rats were raised in the SPF animal room of Yunnan University of Traditional Chinese Me…

Representative Results

Selective spinal manipulation can promote body weight gain in infant rats with cerebral palsy. During the treatment, body weight was measured on postnatal days 3, 14, 28, 42, and 61 (Figure 4, Table 3). On the third day after birth, the body weight of the Sham group was 5.53 ± 0.035 g, and the body weight of the Control group was 3.15 ± 0.43 g. The body weight of the Treatment group was 4.42 ± 0.13 g, and it was not significantly differe…

Discussion

Because ischemia and hypoxia are important pathogenic factors of cerebral palsy, the internationally recognized method of establishing the cerebral palsy model is combined with hypoxia to prepare the cerebral palsy model16,28,29,30. When cerebral palsy develops, it causes global developmental delay, including weight and motor function, which may be related to the level of growth hormone secreti…

開示

The authors have nothing to disclose.

Acknowledgements

This work has been supported by the General Program of National Natural Science Foundation of China (82374614), the Major Biomedical Project of the Science and Technology Department of Yunnan Province (202102AA100016), the Joint Major Project of Applied Basic Research of the Department of Science and Technology of Yunnan Province — Yunnan University of Traditional Chinese Medicine (201901AI070004), Supported by the Key Laboratory of Acupuncture, Moxibustion, and Massage for Prevention and Treatment of Encephalopathy in Colleges and Universities of Yunnan Province (2019YGZ04), Department of Science and Technology of Yunnan Province — Youth Project of Basic Research Program of Yunnan Province (202101AU070002), Graduate Program of Science Research Fund of Education Department of Yunnan Province, (2023Y0433); Scientific Research Foundation of Education Department of Yunnan Province, (2023Y0462).

Materials

96-well plates Beijing Lanjieke Biotechnology Co., LTD 11510 Determination of protein concentration
Anti-beta Actin antibody Abacm Ab8227 Dilution: 1/2000
Anti-Growth Hormone antibody Abacm Ab126882 0.5 µg (0.5 ng/lane)
Anti-Growth Hormone receptor antibody Abacm Ab202964 Dilution: 1/1000
Basic operating microscope Shanghai YuYAN Scientific Instrument Co. LTD SM-101 The common carotid artery was isolated under microscope during modeling
BCA developer Biyuntian Biological Engineering Co., LTD P0010 Determination of protein concentration
Chemiluminescence imaging system Shanghai Qinxiang Scientific Instrument Co., LTD 100240073 Protein banding imaging
Direct-load Color Prestained Marker Beijing Kangrunchengye Biotechnology Co., LTD (GenStar) M221 Western Blot
DK-30Automatic snow ice maker Henan Brothers instrument equipment Co., LTD SHDX0023 Ice-making
ECL luminescent substrate kit Beijing Lanjieke Biotechnology Co., LTD BL520B Convert latent images in exposed film into visible images
Electric-heated thermostatic water bath TAISITE INSTRUMENT DK-98-II The young rats were resuscitated after modeling
Electronic scales Kunshan YoukeWEI ELECTRONIC Technology Co. LTD CN-LQC10002 The body weight of the young rats was measured
German small white electric coagulation pen hemostat Haohang L55×W125×H37 It was used to coagulate the left common carotid artery
Glove Jiangsu YANGzi LiDE Medical Device Co. LTD Q/320684 YZYL001-2017 For massage operation
Glycine Beijing Soleibao Technology Co., Ltd. Cat#G8200 Electrophoretic solution, Configure the transfer fluid
Goat Anti-RabbitIgG H&L (HRP)  Abacm Ab6721 Dilution: 1/10000
Intelligent laboratory ultra-pure water machine Chongqing huachuag water treatment engineering co.,LTD N/A Filtration (15 L)
Isoflurane Shandong Ante Animal Husbandry Technology Co. LTD 15198 Anesthesia was maintained by induction in young rats
LinkIR FOTIRC V1.3.2.134 Infrared image analysis software
Low temperature high speed tissue grinder Wuhan Servicebio technology CO.,LTD SKZ3F20200191 Tissue grinding
Methanol Guangdong Guanghua Sci-Tech Co., Ltd 20220519 Configure the transfer fluid
Mini-PROTEAN Tetra Bole Life Medical Products (Shanghai) Co., Ltd 552BR 233193 Electrophoresis
Multiskan Spectrum Microplate Spectrophotometer TECAN Spark The absorbance and concentration of tissue protein were detected
Pressure-sensing smart gloves Jinan Super Sense Intelligent Technology Co. LTD Miiglove It is used to measure the manipulative strength of the operator
PVDF membrane MerckMillipore Corporation  IPVH00010  Western Blot
Refrigerated centrifuge Hettich Precision Technology (Zhuhai) Co., LTD MIKRO 220R Centrifuge
Research three-in-one thermal imager FOTIRC 226S (384 x 288) Temperature measurement
RIPA lysate Beijing Solaibao Technology Co., LTD (Solarbio) R0010 Lytic tissue
SHA-CA digital display water bath thermostatic oscillator Changzhou Aohua Instrument Co. LTD SHA-CA Young rats were used in hypoxia
Six-rat fatigue rotarod apparatus Shanghai Duoyi Industry Co., LTD DO01104RT703 CP motor function was detected
Skim milk powder Guangzhou Saiguo Biotechnology Co., LTD (BIOFROXX) 1172GR500 Confining liquid
Surgical plate Shanghai YuYAN Scientific Instrument Co. LTD 51002 The model operating table was established in young rats
TS-200 Orbital shaker Haimen Qilin Bell Manufacturing Co., Ltd. TS-8S Gel fixation
Tween 80 MedChemExpress HY-Y1819 Configure TBST 
ZS-MV Portable anesthesia machine ZHONGSHI SCIENCE &TECHNOLOGY ZS-MV-I Anesthesia was induced and maintained in experimental animals
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タグ

Cerebral PalsySelective Spinal ManipulationInfant RatsWeightRotarod TestFoot-fault ScoreGrowth HormoneTraditional Chinese MedicineMassageExternal Therapy
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Gao, T., Wang, Y., Li, M., Yang, L., Chen, S., Gao, L., Shi, Y., Zhang, X., Tai, X. Enhancing the Development and Growth of Infant Cerebral Palsy Rats Using Selective Spinal Manipulations. J. Vis. Exp. (204), e65659, doi:10.3791/65659 (2024).
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