Exploration On The Role Of Hydrogen Sulfide In The Pathogenesis And Treatment Of Achalasia

AimsTo study the expression of hydrogen sulfide synthase in esophageal tissue of patients with achalasia,the effect and possible mechanism of exogenous hydrogen sulfide in the treatment of achalasia animal model,and the treatment of hydrogen sulfide donor(Fluimucil)in patients with achalasia.To explore the role of hydrogen sulfide in the pathogenesis of achalasia and the feasibility of hydrogen sulfide for the treatment of achalasia.Methods(1)Eighteen patients with achalasia and eight patients with squamous cell esophageal cancer were randomly selected as the experimental group and the control group.In the experimental group,the esophageal tissue specimens were obtained in the POEM procedure,and the control group were obtained in the esophagectomy.The experimental group specimens were taken from 4cm above the cardia,2cm above the cardia,LES and the cardia;the control group specimens were took from the LES whose size was same as the experimental group.To observe the expression of CBS and CSE,the samples obtained from the experimental group and the control group were subjected to immunohistochemistry and WB experiments of synthase CBS and CSE and immunofluorescence staining experiment of the two enzymes respectively with sox-10,the neurospecific antibody;(2)Thirty Balb/c mice were randomly divided into model group and control group,the initial body weight was recorded.The model group was given BAC through laparotomy which was injected into the LES and the control group was injected with normal saline after laparotomy.The basic conditions and body weight changes of the postoperative animals were observed and the LES pressure and esophagography were measured again on the 21 st day to evaluate the success of achalasia model.Esophageal tissues taken from all groups were subjected to immunohistochemistry of actin and elastin protein.Another 30 Balb/c mice were randomly divided into three groups: model group,treatment group and control group.The model group was treated with the BAC.Then PBS was intraperitoneally injected for 21 days;the treatment group was treated with the same BAC method as above,and the sodium hydrosulfide solution(14 μmol/kg)was given intraperitoneally for 21 days;the control group was injected with normal saline after laparotomy,and intraperitoneally injected PBS solution for 21 days.The body weights and the LES pressure values of the mice were recorded and esophageal tissues taken from all groups were subjected to immunohistochemistry of actin and elastin protein after 21 days of treatment to observe the changes of the above indicators;(3)Twenty patients with achalasia from Tianjin Medical University General Hospital were randomly selected according to the corresponding inclusion criteria(age,gender,past history,informed consent,etc.)and exclusion criteria(basic status,complications,alergy history,etc).Fluimucil was given orally for 3 months with 200 mg 30 minutes before meals each time and 3 times a day.Patients were followed up monthly.LES pressure values and Eckardt symptom scores were recorded during initial enrollment and the follow-up procedure,and esophageal barium swalow angiography was performed at the initial and the end of the follow-up procedure.Results(1)The results of immunohistochemistry and WB showed that the expression of CBS and CSE in the LES of the patients with achalasia was significantly lower than that of the control group(P<0.05),and the expression of CBS and CSE in the LES of experimental group was significantly lower than that of other parts of the esophagus(P<0.05);Fluorescence staining showed that CBS and CSE enzymes were completely co-expressed with sox-10,neuro-specific antibody and the fluorescence count of LES in the experimental group was significantly less than that of the middle esophagus in the experimental group(P<0.05);(2)The weight gain rate of the achalasia animal model was slower than that of the control group,and LES pressure was significantly increased(P<0.05)and bird’s mouth sign was clearly seen,which means the model was successfully established.The immunohistochemistry results of elastin and actin protein showed that the protein expression of the model group was increased.After intraperitoneal treatment with the sodium hydrosulfide solution,the weight gain increased,the LES pressure was reduced,and the expression of elastin and actin protein in the esophageal tissues was decreased.Whereas,these indicators in treatment group were still higher than those in the control group,and the data differences between the three groups were statistically significant(P<0.05);(3)Seventeen patients were finally enrolled in the Fluimucil experiment,and the success rate after three months of treatment was 70.59%,the symptom relief rate was 82.35%.The patients’ initial LES pressure was(50.06±7.03)mmHg and the initial Eckardt symptom score was(8.12±2.17).After treatment,the patients’ LES pressure was(32.12±6.96)mmHg and Eckardt symptom score was(3.76±2.24).As for the esophageal barium swalowing,the height of the barium was reduced compared with the initial examination,and the esophageal stenosis was relieved.ConclusionsThe expression of CBS and CSE in the esophageal tissue of patients with achalasia was exsited and completely co-expressed with sox-10,the neuro-specific antibody.The expression of CBS and CSE in the LES of esophageal tissue in patients with achalasia was significantly lower than that in the normal control group,and the expression of the above two enzymes in LES was significantly lower than that in the middle esophagus.Animal models and patients of achalasia recovered to some extent after treatment with hydrogen sulfide,which may be related to improving esophageal remodeling.Hydrogen sulfide is reduced in the pathogenesis of achalasia,which may be related to nerve damage.Hydrogen sulfide can be used as a new treatment for achalasia,which provides a new direction for the treatment of this disease.