| Purpose:Objective To observe the effect of electroacupuncture at "Zusanli" and "Sanyinjiao" on the expression of inflammation related genes and proteins in myocardial cells of hypertensive and obese rats,and to explore the mechanism of electroacupuncture at "Zusanli" and "Sanyinjiao" in improving the inflammation of myocardial cells of hypertensive and obese rats from the perspective of NF-?B/NLRP3 signaling pathway.Material and method:In the normal group,10 6-week-old Wistar Kyoto rats(WKY rats),male,weighing 200gą10g,were used;Forty 6-week-old spontaneously hypertensive rats(SHR),male,weighing 200gą10g,were randomly divided into model group,obesity model group,acupuncture group and acupuncture control group with 10 rats in each group.After a week of adaptive feeding,the normal group and model group were fed with normal diet;The obesity model group,acupuncture group and acupuncture control group were fed with high-fat diet.The blood pressure of rats in each group was monitored by intelligent non-invasive sphygmomanometer twice a week.The weight of rats in each group was monitored by intelligent electronic scale twice a week.When the weight of rats in obesity model group,acupuncture group and acupuncture control group exceeded 20% of that in normal group,the model was established successfully.After successful modeling,the acupuncture group was treated with filiform needle with a depth of about 5mm at bilateral "Zusanli" and "Sanyinjiao" points;In the acupuncture control group,the upper 1/3(non meridian and non acupoint)of the tail of rats was selected as the control;The model group and obesity model group were fixed daily without acupuncture.Acupuncture group and acupuncture control group: the needle handle was connected with electroacupuncture instrument.Intensity: 1m A,with slight limb tremor as the degree.Frequency: sparse wave,sparse wave: 1Hz,dense wave:5Hz,sparse wave time: 10 s,dense wave time:15s.20 minutes each time,acupuncture once a day,continuous treatment for 4 weeks.After the treatment,rats were fasted for24 hours and injected intraperitoneally with 10% chloral hydrate solution at a dose of0.4ml/100 g.After anesthesia,the rats were fixed on the experimental platform.Part of the heart tissue was taken and placed in a sterile EP tube and stored in a refrigerator at-80? for standby.Part of the heart was taken and fixed in 4% paraformaldehyde.The expression of NF-?B m RNA was measured by fluorescence quantitative PCR,the protein expressions of NF-?B,NLRP3,caspase-1 and IL-1? were measured by WB;HE staining was used to observe the heart inflammation.The experimental data were analyzed by spss26.0 statistical software,Use the meanąstandard deviation(SX(10)),One way ANOVA test was used to analyze the pairwise comparison between groups.LSD method was used to test the hypothesis of equal variance between groups.Tamhani T2(M)test was used to test the hypothesis of equal variance between groups.The difference was statistically significant(P<0.05).Results:1.The weight changes of rats in each group were as follows: With the beginning of acupuncture treatment,the weight of the acupuncture group began to decrease gradually,and gradually approached the normal group;the body weight of the acupuncture control group showed a normal growth range.After four weeks of acupuncture treatment,compared with the normal group,the weight of rats in the obesity model group was significantly higher than that in the normal group(P<0.05);compared with the model group,the weight of the obese model group was significantly higher than that of the model group(P<0.05);the body weight of the acupuncture group was significantly lower than that of the obesity model group(P<0.05);the difference between the acupuncture control group and the acupuncture group was statistically significant(P<0.05).2.The changes of blood pressure of rats in each group were as follows: With the beginning of acupuncture treatment,the blood pressure of the acupuncture group began to decrease gradually,and gradually approached the normal group;there was no significant change in blood pressure in the acupuncture control group.After four weeks of acupuncture treatment,compared with the normal group,the weight of rats in the model group,obesity model group and acupuncture control group was significantly higher than that in the normal group(P<0.05);compared with the model group,the weight of obesity model group and acupuncture control group was significantly higher than that of model group,and the weight of acupuncture group was significantly lower than that of model group(P<0.05);the body weight of the acupuncture group was significantly lower than that of the obesity model group(P<0.05);the difference between the acupuncture control group and the acupuncture group was statistically significant(P<0.05).3.HE staining results showed that:He staining showed that the morphology and structure of myocardial cells in normal group were clear,the intercellular space was small,the structure of myocardial fibers was complete,and there was no obvious inflammatory cell infiltration;In the model group and the obesity model group,the myocardial morphology and structure were not clear,the arrangement was disordered,the myocardial gap was enlarged,accompanied by extensive inflammatory cell infiltration,and the degree of inflammation in the obesity model group was significantly heavier than that in the model group;In the acupuncture group,the pathological changes of myocardial tissue were significantly alleviated,the cell morphology was clear,the arrangement was relatively regular,and the infiltration of inflammatory cells was alleviated;The inflammation in the acupuncture control group was not relieved.4.The protein expression of rat heart in each group was statistically analyzed: Compared with the normal group,the NF-?B,NLRP3,Caspase-1 and IL-1? protein content in the model group was significantly increased(P<0.05);Compared with the model group,the content of NF-?B,NLRP3,Caspase-1 and IL-1? protein in the obesity model group was significantly increased(P<0.05);Compared with the obesity model group,the NF-?B,NLRP3,Caspase-1and IL-1? protein content in the acupuncture group was significantly lower(P<0.05);The NF-?B,NLRP3,Caspase-1 and IL-1? protein content in the acupuncture control group was significantly higher than that in the acupuncture group(P<0.05).5.According to the statistical results,the m RNA expression of rat heart in each group was as follows: Compared with the normal group,the expression of NF m RNA in the model group was significantly higher than that in the normal group(P<0.05);Compared with the model group,the expression of NF m RNA in the obesity model group was significantly higher than that in the model group(P<0.05);The expression of NF m RNA in the acupuncture group was significantly lower than that in the obesity model group(P<0.05).Conclusion:Electroacupuncture at "Zusanli" and "Sanyinjiao" can regulate NF-?B/NLRP3 signaling pathway,reduce the expression of IL and other inflammatory factors,This proves that electroacupuncture can protect the normal operation of heart function in patients with hypertension and obesity. |
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