| Objective:In this study,functional dyspepsia(FD)rat was established to study the effect of massage on gastric motility in FD rats,and to clarify the improvement effect of massage on FD.Based on the mechanism of oxidative stress,this paper further studies the mechanism of“Tongjing Tiaozang” massage therapy in the treatment of FD from the perspective of repairing mitochondrial damage,regulating mitophagy,and improving gastric motility,and reveals the internal relationship and mechanism between its effect and oxidative stress,so as to provide experimental basis and theoretical basis for the clinical promotion of “Tongjing Tiaozang” massage therapy.Methods:1.Effect of massage method of “Tongjing Tiaozang” on the general condition of FD ratsIn this experiment,FD rat was constructed by irregular diet combined with tail clamping stimulation.In the treatment,the massage method of “Tongjing Tiaozang” was used as the treatment group;the gastrointestinal motility drug mosapride was used as the positive drug control group,and a normal control group was set up.During the experiment,the daily state of rats in each group was observed and recorded.Body weight,diet and water intake were monitored regularly.Through the above research,the effect of “Tongjing Tiaozang” massage on the general condition of FD rats was clarified.2.Effects of “Tongjing Tiaozang” massage on gastric motility in FD ratsAfter the experiment,the gastric emptying rate of rats in each group was detected.The expression of c-kit and Cx43 in gastric tissue was detected by Western Blot,q PCR and immunohistochemistry.The content of ATP in gastric tissue was detected by ELISA method.Through the above research,the effect of massage on gastric motility in FD rats was clarified.3.Effects of “Tongjing Tiaozang” massage on oxidative stress in gastric tissue of FD ratsThe content of ROS in gastric tissue was detected by ELISA,the content of MDA and the activity of SOD in gastric tissue were detected by colorimetry,and the effect of“Tongjing Tiaozang” massage on oxidative stress in gastric tissue of FD rats was determined.4.Effects of “Tongjing Tiaozang” massage on mitochondrial damage in gastric tissue of FD ratsThe mitochondria of gastric tissue were observed by transmission electron microscopy.The expression level of AMPK-ULK1 signaling pathway in gastric tissue was detected by Western Blot.To clarify the effect of “Tongjing Tiaozang” massage on mitochondrial damage in FD rats.5.Effect of “Tongjing Tiaozang” massage on mitophagy in FD ratsThe co-expression of LC3 and VDAC1 in gastric tissue was detected by immunofluorescence.The expression levels of autophagy-related proteins P62 and LC3II/I and mitophagy pathway PINK1-Parkin signaling pathway in rat gastric tissue were detected by Western Blot.Through the above research,the effect of “Tongjing Tiaozang” massage on mitophagy in FD rats was clarified.Results:1.Effect of “Tongjing Tiaozang” massage on the General Condition of FD ratsAfter the experiment,the model group was depressed and unresponsive,the rat hair was messy and yellow,the feces were yellow-green loose stool,and the diet,water intake and body weight decreased significantly.Compared with the model group,the mental state of the massage group gradually improved,the activity increased significantly,the rat hair was not much different from the normal group,the feces were formed,the quality was slightly soft,and the diet,water intake and body weight increased significantly.Compared with the massage group,the mosapride group was more depressed,with less activity,and there was no significant difference in other aspects.It is suggested that the massage method of “Tongjing Tiaozang” has a significant improvement effect on the general state of FD rats.At the same time,the results of HE staining showed that there were no organic lesions such as inflammation and ulcers in the gastric tissues of rats in each group,indicating that the FD rat model was successfully prepared.2.Effects of “Tongjing Tiaozang” massage on gastric motility in FD ratsGastric emptying detection is the most direct detection method of gastric motility.The results showed that compared with the normal group,the gastric emptying rate of the model group decreased significantly(P<0.01),suggesting that the gastric motility of the model group decreased under FD state.The gastric emptying rate of the massage group was significantly higher than that of the model group(P<0.01).It shows that the massage method of “Tongjing Tiaozang” can restore the gastric motility of FD rats and improve the disease state of FD.ICCs are pacemaker cells in the gastrointestinal tract,and their abnormal number and network structure are one of the important factors affecting gastric motility.C-kit is the specific receptor of ICCs.Cx43 is a gap junction protein connecting ICCs and smooth muscle.The content of both can reflect the number and network structure of ICCs.The results showed that compared with the normal group,the expression of Cx43 and c-kit in the model group decreased significantly,suggesting that the number and network structure of ICCs in the model group were abnormal in FD state.The expression of Cx43 and c-kit in the massage group was significantly higher than that in the model group,and better than that in the mosapride group.The above results indicate that the massage method of “Tongjing Tiaozang” can restore gastric motility and improve the disease status of FD by maintaining the number and network structure of ICCs in gastric antrum tissue.ATP is the energy supply of gastrointestinal motility,and the decrease of energy supply can affect the gastric motility of FD rats.The results showed that compared with the normal group,the ATP content in the model group decreased significantly(P<0.01),suggesting that the energy supply of gastrointestinal motility in FD state decreased.The ATP content in the massage group was significantly higher than that in the model group(P<0.05),indicating that the massage method of “Tongjing Tiaozang” can inhibit the decrease of energy supply in the gastric tissue of FD rats,restore gastric motility,and improve the disease status of FD.3.Effects of “Tongjing Tiaozang” massage on oxidative stress in FD ratsExcessive production of ROS is the main cause of oxidative stress.SOD and MDA can be used as markers to evaluate this stress level,which can directly reflect the degree of oxidative damage.The results showed that compared with the normal group,the contents of ROS and MDA in the model group increased significantly(P<0.01),and the activity of SOD decreased significantly(P<0.01).Compared with the model group,the content of ROS and MDA in the massage group decreased significantly(P<0.01)and the activity of SOD increased significantly(P<0.05).It shows that the massage method of “Tongjing Tiaozang”can improve the oxidative stress state.The results of transmission electron microscopy of gastric tissue showed that compared with the normal group,the number of mitochondria in the model group decreased,and more damaged mitochondria and autophagosomes could be seen.Compared with the model group,the number of mitochondria in the massage group increased significantly,and the number of damaged mitochondria and autophagosomes decreased.It shows that “Tongjing Tiaozang”massage can alleviate mitochondrial damage caused by oxidative stress.AMPK-ULK1 is an important signaling pathway that causes autophagy.The results showed that compared with the normal group,the expression of phosphorylated AMPK and ULK1 protein in the model group increased significantly(P<0.01),suggesting that mitochondrial damage caused by oxidative stress can cause abnormal autophagy.The expression of phosphorylated AMPK and ULK1 protein in the massage group was significantly lower than that in the model group(P<0.05),which was basically the same as that in the mosapride group.The above results indicate that “Tongjing Tiaozang” massage can reduce the occurrence of abnormal autophagy by inhibiting the activation of AMPK-ULK1 signaling pathway.4.Effect of “Tongjing Tiaozang” massage on mitophagy in gastric tissue of FD ratsP62 and LC3 are the marker proteins of autophagy.When autophagy occurs,the expression of P62 decreases and the ratio of LC3II/I increases.At the same time,the co-expression of LC3 and VDAC1 can reflect the level of mitophagy.The results showed that compared with the normal group,the expression level of P62 in the model group decreased significantly,the ratio of LC3 II/I increased significantly,and the co-expression area of LC3 and VDAC1 increased significantly(P<0.01),suggesting that the FD model showed abnormal mitophagy.Compared with the model group,the expression level of P62 in the massage group was significantly increased,the ratio of LC3 II/I was significantly decreased,and the co-expression area of LC3 and VDAC1 was significantly decreased(P<0.01).The above results indicate that “Tongjing Tiaozang” massage can reduce the occurrence of abnormal mitochondrial autophagy..In the PINK1-Parkin signaling pathway: Compared with the normal group,the expression levels of PINK1 and Parkin in the model group were significantly increased(P<0.01);compared with the model group,the expression levels of PINK1 and Parkin in the massage group and the mosapride group were significantly decreased(P<0.05).It shows that the “Tongjing Tiaozang” massage method can alleviate mitochondrial excessive autophagy by regulating the PINK1-Parkin signaling pathway.Conclusion:1.The massage method of “Tongjing Tiaozang” can not only improve the mental state,activity,hair and defecation of FD rats,but also increase the diet,water intake and body weight,and improve the general state of FD rats.2.The massage method of “Tongjing Tiaozang” can maintain the number and network structure of ICCs by increasing the expression levels of Cx43 and c-kit in gastric antrum tissue,increase the content of ATP,increase the energy supply of gastric tissue,and then restore the gastric motility and improve the gastric emptying ability of FD rats.3.The massage method of “Tongjing Tiaozang” can reduce the content of ROS in gastric tissue,increase the activity of SOD,reduce the release of MDA,and thus alleviate the oxidative stress state of gastric tissue.4.The massage method of “Tongjing Tiaozang” can reduce the mitochondrial damage of gastric tissue caused by oxidative stress,inhibit the activation of AMPK-ULK1 signaling pathway,and reduce the occurrence of abnormal autophagy.5.The massage method of “Tongjing Tiaozang” inhibiting the activation of AMPK-ULK1 signaling pathway in gastric tissue,thereby inhibiting the activation of PINK1-Parkin signaling pathway,the abnormal mitophagy was improved.6.The massage method of “Tongjing Tiaozang” can enhance gastric motility and improve FD by alleviating oxidative stress,repairing mitochondrial damage and regulating mitophagy in gastric tissue. |
PDF Full Text Request