| ObjectiveTo explore the different effects of reinforcing and reducing methods by twirling and rotating the needle on prevention for left ventricular damage of SHR.. the blood pressure in SHR,endothelin-1,the level of Ⅰ、Ⅲ collagen mRNA, production of eNOs and gene expression of P38/MAPKin left ventriclewere measured by different biochemistry methods to evaluatecurative effects of acupuncture. Observing the morphology of left ventricle of SHR aims to reveal weather acupuncture has a good effect on preventingleft ventricular damage of SHR. Which can also help to reveal the mechanism in molecular level, providing scientific and objective evidence for different effects of reinforcing and reducing methods by twirling and rotating the needle.MethodSixty male SHR of11weels’old were randomly divided into four groups:model group (A), group (B) with twirling reinforcing method, group (C) with twirling reducing method, group (D) without manipulation, and a normal Wistar rats of the same age with SHR as group (E) without acupuncture. Group B、 C、D are all treated on single "Taichong" point (LR3) every day, ten minutes every time, rest one day every six days.28days later observing myocardial hypertrophy and myocardial fibrosis situation of each group with biochemistry methods. As well as their blood pressure change, observing tissues of HE and Masson staining of left ventricleunder light microscopy, testing the content of endothelin-1by ELISA, detecting the level of of I and Ⅲ collagen mRNA, observing immunohistochemical with cardiac eNOs and gene expression of P38/MAPK.ResultsThe changes of systolic pressure and diastolic pressure of each groupBefore the experiment the average blood pressure of SHR groups has no statistic difference among each group, but higer than blank control group E(P<0.05). After experiment the systolic pressure and diastolic pressure of group A and group B are higher than other groups, no significant difference between A and B(p>0.05); compared with group C and group D, revealing statistic differences (p<0.05), the blood pressure are lower in both group C and D,However the blood pressure of group C is much lower than group D(P<0.05). the blood pressure of SHR groups are all still in accordance with the standard of hypertension compared with Wistar group.The results of HE and Masson stainingHE and Masson pictures show myocardial hypertrophy and myocardial fibrosis on group A、B、C and D. but less serious on group C, which were characterized by disorder of myocardial cells, bigger unclear, collagen hyperplasia, vascular changes and collagen fibers deposition on endocardium of left ventricle. Group E is normal.The content of endothelin-1of each groupThe content of endothelin-1of each group is different from each other(P<0.05), content of five group ranked from high to low are:A>B>D>C>E.Level of Ⅰ、Ⅲ collagen mRNA of each groupSemi quantitative PCR of I collagen mRNA shows that there is no statistic difference between group C and group D (P>0.05), but the both groups compared with group A and B, showing significant difference (P<0.05), the level of group C and D is lower than group A and B;similarly, the level of Ⅲ collagen mRNA shows that group C is lower than group A, B and D (P<0.05), but higher than normal group E (P<0.05).The result of immunohistochemical analysis with eNOsThe analysis shows that all five groups appear positive eNOsmolecules, mainly distributed in vascular walls and vascular cavity of left ventricle,compared with other groups, acupuncture with reducing method group and acupuncture without manipulation group show more positive eNOs, but there is no significant difference between the two groups(P>0.05).Western blot analysis of P38/MAPKThe result of P38enzyme shows that model group A has the strongest activity (P<0.05), compared with other groups. Group E is stronger than group C and D, but weaker than group B(P<0.05), the activity of group D is the weakest.Conclusion(1) Acupuncture with twirling reducing method and acupuncture without manipulation method can effectively decrease the blood pressure of SHR, however, reducing method is more effective, but twisting reinforcing method has no effect on controlling blood pressure.(2) Twirling reducing method and acupuncture without manipulation method can both effectively suppress the production of endothelin-1, level of collagen I and III in left ventricle of SHR, but twirling reducing method plays a more effective role on prevention. However the experiment shows that acupuncture can depress the gene expression of P38, acupuncture without manipulation has the most effective inhibiting action, secondly the reducing method and thirdly the reinforcing method.(3) Reducing metod and acupuncture without manipulation method both can promote the produnction of eNOs in left ventricle, but no significant statistic difference between the two methods. (4) Twirling reducing method has significant effect on preventing myocardial hypertrophy and myocardial fibrosis, which may work through decreasing the biological activity of myocardial P38, then reducing the production of ET-1and the level of collagen Ⅰ and Ⅲ mRNA further. On the other hand increasing the expression of eNOs in left ventricle then to improve the supply of oxygen of myocardial cells, moreover to relax the vascular smooth muscle.(5) Twirling reducing method has an important role on preventing myocardial damage caused by hypertension. Acupuncture without manipulation can also protect myocardial cells, but less effective, however reinforcing method neither has protection effect nor damage effect. Therefore, twirling reducing and twirling reinforcing method have different biomedical effect on16weeks’SHR. |
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