Study On The Effect Of Ghrelin/GHS-R Expression In CP Rats Through LD Ointment Tuina

Objective:The effects of LD Ointment Tuina on the growth and development of CP rats were observed by detecting the changes in the expression of Ghrelin and GHS-R in CP rats,and to explore the molecular mechanism of LD Ointment Tuina on the growth and development of CP rats.So that can lay a foundation for the clinical application of the invention patent of LD Ointment.Method:108 newborn SD rats were screened strictly according to the experimental criteria.Female and high or low weight rats were excluded.59 male rats were randomly divided into Model Group(n=39),Sham Control Group(n=10),Blank Control Group(n=10).The Model Group was prepared by left carotid artery ligation combined with hypoxia,while the left carotid artery was separated in the Sham Control Group without ligation and the Blank Control Group without operation.30 SD rats with successful modeling were randomly divided into three subgroups: LD Ointment Tuina Group(n=10),Oil Control Group(n=10),Model Control Group(n=10).The LD Ointment Tuina Group and the Oil Control Group received tuina intervention.LD Ointment Tuina Group used LD Ointment as the medium.Oil Control Group used the oil as the medium.Body weight,body length,tail length and eyes-open time were recorded at the same time points.Praxiology experiment were tested at the same time points.Detecting the changes of Ghrelin and GHS-R in blood and brain tissue.The data were statistically analyzed by one-way ANOVA and Independent Samples T-Test.When P < 0.05(P < 0.01),was statistically significant.Result:1.After modeling,the model was verified by the righting reflex experiment.The average time of the Model Group was 12.550±5.287 s,the Sham Control Group was1.340±0.323 s,and the Blank Control Group was 1.436±0.209 s.Compared with Sham Control Group and Blank Control Group,the Model Group had significant difference(P <0.05).2.After 4 weeks of tuina intervention,the average weight of LD Ointment Tuina Group was 114.92±2.79 g,the Oil Control Group was 99.93±4.43 g,the Model Control Group was 85.96±3.23 g,the Sham Control Group was 111.71±3.17 g,and the Blank Control Group was 111.77±3.68 g.Compared with Oil Control Group,Model Control Group and Blank Control Group,the difference was statistically significant(P < 0.05).After 4 weeks of tuina intervention,the average length of body and tail was 30.83±0.61 cm in LD Ointment Tuina Group,28.78±0.62 cm in Oil Control Group,26.50±0.79 cm in Model Control Group,30.75±0.53 cm in Sham Control Group and 30.71±0.72 cm in Blank Control Group.Compared with Oil Control Group and Model Control Group,the difference was statistically significant(P < 0.05).After 4 weeks of tuina intervention,the average time of eye opening was 14.2±0.42 days in LD Ointment Tuina Group,15±1.05 days in Oil Control Group,16.4±1.58 days in Model Control Group,11.9±0.32 days in Sham Control Group and 11.8±0.42 days in Blank Control Group.Compared with Oil Control Group,Model Control Group and Blank Control Group,the difference was statistically significant(P < 0.05).3.After 4 weeks of tuina intervention,the average gait analysis motion speed of LD Ointment Tuina Group was 35.725±1.626 cm/s,the Oil Control Group was 30.450±1.237 cm/s,the Model Control Group was 24.148±0.701 cm/s,the Sham Control Group was 35.909±1.169 cm/s,and the Blank Control Group was 35.701±0.875 cm/s.Compared with Oil Control Group and Model Control Group,the difference was statistically significant(P < 0.05).After 4 weeks of tuina intervention,the average duration of gait analysis feet support was 0.150±0.020 s in LD Ointment Tuina Group,0.132±0.009 s in Oil Control Group,0.112±0.010 s in Model Control Group,0.153±0.015 s in sham control group and 0.157±0.008 s in Blank Control Group.Compared with Oil Control Group and Model Control Group,the difference was statistically significant(P < 0.05).After 4 weeks of tuina intervention,the average number of fatigue motion analysis was 297.2±7.01 s for the third time in LD Ointment Tuina Group,245.8±83.15 s in Oil Control Group,77.9±37.34 s in Model Control Group,300±0.00 s in Sham Control Group and 300±0.00 s in Blank Control Group.Compared with the Oil Control Group,the Model Control Group,the difference was statistically significant(P < 0.05).4.After 4 weeks of tuina intervention,Ghrelin expression in the peripheral of LD Ointment Tuina Group averaged 43.155±6.035 pg/ml,the Oil Control Group averaged41.070±2.771 pg/ml,the Model Control Group averaged 35.462±4.08 pg/ml,the Sham Control Group averaged 39.916±2.935 pg/ml,and the Blank Control Group averaged39.893±2.325 pg/ml.Compared with the Model Control Group,the difference was statistically significant(P < 0.05).After 4 weeks of tuina intervention,the peripheral GHS-R expression in the LD Ointment Tuina Group averaged 8.487±0.457 ng/ml,the Oil Control Group averaged8.079±0.884 ng/ml,the Model Control Group averaged 6.745±0.985 ng/ml,the Sham Control Group averaged 7.863±1.476 ng/ml,and the Blank Control Group averaged 7.873±0.768 ng/ml.Compared with the Model Control Group,the difference was statistically significant(P < 0.05).After 4 weeks of tuina intervention,the average ratio of central Ghrelin expression was 1.007±0.222 in the LD Ointment Tuina Group,0.792±0.216 in the Oil ControlGroup,0.215±0.111 in the Model Control Group,0.487±0.215 in the Sham Control Group and 0.495±0.230 in the Blank Control Group.Compared with the Model Control Group and the Blank Control Group,the difference was statistically significant(P < 0.05).After 4 weeks of tuina intervention,the average ratio of central GHS-R expression was 0.562±0.161 in the LD Ointment Tuina Group,0.383±0.118 in the Oil Control Group,0.110±0.067 in the Model Control Group,0.224±0.114 in the Sham Control Group and 0.221±0.108 in the Blank Control Group.Compared with the Model Control Group and the Blank Control Group,the difference was statistically significant(P < 0.05).Conclusion:1.LD Ointment Tuina could promote the growth and development of CP rats,and the experimental results were consistent with previous literature reports.2.LD Ointment Tuina could improve the motor balance function of CP rats,and the experimental results were consistent with previous literature reports.3.The effect of LD Ointment Tuina on promoting the growth and development of CP rats and improving the motor balance function may be achieved by affecting the protein expression of Ghrelin and GHS-R and regulating the release of GH.