| ObjectiveBased on twirling reinforcing and reducing manipulation on spontaneously hypertensive rats (SHR)to observe the changes of blod pressure (systolic blod pressure/diastolic blood pressure,SBP/DBP), concentration of serum Angiotensin II (Ang II) and abdominal aorta matrix metalloproteinase-9(MMP-9), mRNA expression of pro collagen I (PC I)*procollagen Ⅲ (PCⅢ), P38Mitogen Activated Protein Kinases(P38/MAPK). From the point of the effect of twirling reinforcing and reducing manipulation on the P38/MAPK signal transduction pathway, the research aims at exploring the correlation between the reversal mechanism of artery ECM reconstruction of SHR rats and P38/MAPK signal transduction pathway and the differential effects of twirling reinforcing and twirling reducing manipulation. This article aims at revealing the mechanism of different biological effects as well as the differential effects of twirling reinforcing and twirling reducing manipulation, in order to providing objective basis for acupuncture on the reversal mechanism of artery ECM reconstruction of SHR rats as well as practicability, scientificity of twirling reinforcing and reducing manipulationMethodClean grade healthy male aged11weeks15WKY rats and60SHR rats weighing240-280g.15WKY rats as normal control group(A).60SHR rats were randomly divided into4groups: the model control group (B)、acupuncture control group(C)、twirling reinforcing method group(D)、twirling reducing method group(E). After the rats were fed one week,we started the operation. The operation began at AM8:00. Group A and B only with catch, bundle stimulation as other groups do, lasting for10minitues. Group C、D、E are all treated on double "Taichong" point(LR3) the next day alternating, rest one day every six days, total for28days. Group C with simple acupuncture for10minitues. Group D with twirling reinforcing method and group E with twirling reducing method. Twisting range of360°/time, frequency is60/min,1min constant twist,9min every time. Acupuncture operation is performed by the same person every time. Measured the artery systolic and diastolic bfood pressure in rat tails every PM4:00on the day before acupuncture treatment and treatment for the7,14,21,28days.28days after treatment, the rats were decapitated and clipped abdominal aorta. Enzyme linked immunosorbent assay (ELISA) to detect the content of Ang Ⅱ in serum and MMP-9in abdominal aorta. Reverse transcription PCR (RT-PCR) to detect the gene expression of PC I, PC III, P38/MAPK in abdominal aorta.Result1Effect of twirling reinforcing and reducing manipulation on systolic pressure, diastolic pressure of rats in each groupSystolic and diastolic blood pressure of SHR groups were higher than those of WKY (P<0.05), but they have no significant difference in SHR groups(P>0.05). Before and after acupuncture treatment for28days,blood pressure have changes as follows:group A elevated diastolic blood pressure after treatment compared with before treatment (P<0.05), but it had no obvious difference in systolic blood pressure compared before and after treatment (P>0.05); group B increased both systolic and diastolic blood pressure after treatment compared with before treatment (P<0.05); both group C and group E decreased systolic and diastolic blood pressure after treatment compared with before treatment (P<0.05); group D elevated systolic blood pressure after treatment compared with before treatment (P<0.05), but it had no obvious difference in diastolic blood pressure compared before and after treatment (P>0.05). Acupuncture treatment for28days, comparison of blood pressure between groups are as follows:systolic and diastolic blood pressure of SHR groups were higher than those of-WKY (P<0.05); the systolic and diastolic blood pressure of both group E and group C decreased compared with those of group B(P<0.05)and blood pressure of group E was significantly lower; both systolic and diastolic blood pressure of group D had no significant difference compared with those of group B (P>0.05); both systolic and diastolic blood pressure of group E decreased compared with those of group C(P<0.05); both systolic and diastolic blood pressure of group D increased compared with those of group C(P<0.05); both systolic and diastolic blood pressure of group E decreased compared with those of group D(P<0.05).2Effect of twirling reinforcing and reducing manipulation on content of Ang Ⅱ in serum of rats in each groupAng Ⅱ levels of group B, C, D, E were increased compared with that of group A (P<0.05); Ang Ⅱ levels of group C, D, E were decreased compared with that of group B (P<0.05); Ang Ⅱ levels of group C had no significant difference compared with that of group D (P>0.05); Ang Ⅱ levels of group E decreased compared with that of group C,D(P<0.05).3Effect of twirling reinforcing and reducing manipulation on content of MMP-9in abdominal aorta of rats in each groupMMP-9levels of group B, C, D, E were increased compared with that of group A (P<0.05); MMP-9levels of group C, D, E were decreased compared with that of group B (P<0.05); MMP-9levels of group C had no significant difference compared with that of group D (P>0.05); MMP-9levels of group E decreased compared with that of group C,D(P<0.05).4Effect of twirling reinforcing and reducing manipulation on gene expression of PC Ⅰ,PC Ⅲ in abdominal aorta of rats in each groupOn the expression of PC Ⅰ mRNA: expression of PC Ⅰ mRNA of group B, C, D, E were increased compared with that of group A (P<0.05); expression of PC Ⅰ mRNA of group C, D, E were decreased compared with that of group B (P<0.05); among PC Ⅰ mRNA expression of group C,D, E, they had no significant difference (P>0.05).On the expression of PCⅢmRNA:expression of PC Ⅲ mRNA of group B, C, D, E were increased compared with that of group A (P<0.05); expression of PCⅢmRNA of group C, D, E were decreased compared with that of group B (P<0.05); expression of PC Ⅲ mRNA of group C had no significant difference compared with that of group D (P>0.05); expression of PC Ⅲ mRNA of group E decreased compared with that of group C,D(P<0.05).5Effect of twirling reinforcing and reducing manipulation on gene expression of P38/MAPK in abdominal aorta of rats in each groupExpression of P38/MAPK mRNA of group B, C, D, E were increased compared with that of group A (P<0.05); expression of P38/MAPK mRNA of group C, D, E were decreased compared with that of group B (P<0.05); expression of P38/MAPK mRNA of group C had no significant difference compared with that of group D (P>0.05); expression of P38/MAPK mRNA of group E decreased compared with that of group C,D(P<0.05).Conclusion1Acupuncture decreasing blood pressure (SBP and DBP) have certain effect differences, the antihypertensive effect of twirling reducing method is more obvious than simple acupuncture,but twirling reinforcing method can elevate systolic blood pressure.2Acupuncture can reduce the content of Ang Ⅱ in serum,MMP-9in abdominal aorta and the expression of PC Ⅰ,PCⅢ,P38/MAPK mRNA in abdominal aorta of each SHR group. In addition to PC Ⅰ, the twirling reducing method can greatly decrease the content or gene expression of other index,which is better than the twirling reinforcing method as well as simple acupuncture.Twirling reinforcing and reducing method and simple acupuncture had no significant difference in expression of PC Ⅰ mRNA.3The mechanism of simple acupuncture as well as twirling reducing manipulation reversing arterial vascular reconstruction of ECM might be related to the inhibition of P38/MAPK signal transduction pathway and twirling reducing manipulation may be the maximum likelihood. By reducing the content of AngⅡ, the inhibition of P38/MAPK signaling pathway was inhibited. Then it caused the decreased concentration of MMP-9and expression of PC Ⅰ,PC Ⅲ mRNA in arteries,which plays an important role in preventing the excessive deposition of collagen and over decomposition of elastin of the artery wall and eventually reverses artery ECM reconstruction to achieve the antihypertensive effect. In this experiment, twisting reinforcing method for elevating systolic blod pressure,negative correlation with other indexes decreased.So whether twisting reinforcing method being able to reverse artery ECM reconstruction is still need to further research. |
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