| ObjectiveRecently, people have gradually realized the important role of gallbladder motility disturbance in the pathogenesis of gallstone , and realized the effects of gastrointestinal peptide hormones on the contraction and relaxation of gallbladder and Oddi's sphincter. These hormones may alter one or more lithogenic factors and affect the formation of gallstones . It is necessary to study these factors to discuss the etiology of gallstones. The content of MTL and SP in plasma and gallbladder wall tissue of the patients with cholelithiasis was measured to evaluate their role and clinical significance in the formation of gallstones with radioimmur noassay in this study.Materials and methodsI . Materials( I ) Main instrument and apparatus : EK -801 vacum pump FT -613 RIA meter TGL - 16G centrifugor rapid liquid mixture UP - 200H homogenize,SANYO -70℃ refrigerator.SHINKO -SH -210R electric analysis scale.( fl ) Main reagents; aprotinin, 10% EDTA - 2Na MTL RIA kit.SP RIA kit.II. Methods( I ). Subjects1. Gallstones group-, all patients were collected from the 2nd affiliated hospital of China Medical University from 2004.9 to 2004. 12 . gallstones Patients with gastrointestinal heart lung liver kidney diseases were excluded. The oper-ation confirmed the diagnosis of gallstones. 36 patients were enrolled, with 13 males and 23 females. The average age was 57. 8 ± 10.9 years old , with range from 34 to 75 years old. All patients received ultrasonograph study and didn t take drugs that affect bile duct motility in the near future.2. Gallbladder polyps group: 14 patients were enrolled ,with 6 males and 8 females . the average age was 43. 1 ±11.6 years old, with range from 26 to 64 years old. The diagnosis was confirmed by operation and pathology studies. The rest condition were similar to these of gallstones group.3. Common bile duct stone group; 11 patients were enrolled,with 3 males and 8 females. The average age was 59.4 ± 18.7 years old,with range from 21 to 82 years old . The radiology studies confirmed the diagonasis of common bile duct stone. The rest condition were similar to these of gallstones group.4. Control group: Blood samples were collected from healthy doctors and nurses and relatives of patients. 10 samples were collected ,with 8 males and 2 females ,The average age was 45.5 ±14.0,with range from 32 to 71 years old. 7 normal gallbladder wall tissue samples were collected from the donor during liver transplantation.(II). Samples collection1. Blood samples for MTL:2ml vein blood sample was collected from overnight fasting patients in the morning , added into tubes with 30ul 10%EDTA -2Na and 30ul aprotinin, mixed , centrifugated at 4℃ (1500r/min) for 15 min. The supernate were stored at -70℃ for later analysis.2. Blood samples for SP: 3 ml vein blood sample was collected from overnight fasting patients in the morning , added into tubes with 30ul 10% EDTA -2Na and 30ul aprotinin,mixed ,centrifugated at 4℃ (1500r/min)for 15min. The supernate were stored at - 70℃ for later analysis.3. Gallbladder wall tissue samples for MTL:300mg gallbladder wall tissue sample was collected , boiled in lml physiological saline for 3 min . then 0.5ml 0. 1N icy hydrochloric acid was added,mixed,setted at 4℃ 1.5 hours,neutralized with 0. 1N NaOH 0. 5ml and then centrifugated at 4℃ (3000r/min) for 30 min . The upper serum was collected, stored at - 70℃ for later analysis.4. Gallbladder wall tissue samples for SP:300mg gallbladder sample wascollected , weighted and boiled in 1ml physiological saline for 3 min . then 0.5ml IN icy acttric acid was added,mixed,setted at 4℃1. 5 hours,neutralized with IN NaOH 0.5ml and then centrifugated at 4℃ (3000r/min) for 30min . The upper serum was collected, stored at -70℃ for later analysis.( I) MTL and SP were measured by RIA.( IV ) Statistical anaysis: All the datas were analyized by SPSS 11.0 software. The results were expressed as x ± s,and t test was used. Statistical significance was defined as P < 0.05.ResultsI .The results of MTL:1. Results of plasma MTL : The plasma MTL level in gallstone group was higher,compared with that of control and gallbladder polyp group(P < 0. 05,P< 0.05 ). There was no statistical significance between gallbladder polyp group and control group. The same result was found between common bile stone group and control group and gallbladder polyp group.2. Results of gallbladder wall tissue MTL: the gallbladder wall tissue MTL level in gallstone group was higher , compared with that of control and gallbladder polyp group( P < 0.05, P < 0.05 ). There was no statistical significance between gallbladder polyp group and control group.II. The results of SP:1. Results of plasma SP: the plasma SP level in gallstone group was higher, compared with that of control and gallbladder polyp group(P < 0. 05,P < 0. 05). There was no statistical significance between gallbladder polyp group and control group. The plasma SP level in common bile stone group was higher than that of control group.2. The measurment of SP of gallbladder tissue failed and no results were obtained.DiscussionUp to date, more than sixty categories and ten groups of gastrointestinal peptide hormones have been discovered. These hormones play a role in the regulation of extra - hepatic bile duct dynamics. Researches have shown that the changes of the levels of these hormones appear before bile stasis and are in accordance with the changes of gallbladder empty. Thus, it is considered that the change of the gastrointestinal peptide hormones is the cause of bile stasis. This alteration may affect the motility of gallbladder, lead to the disturbance of bile juice excretion and play an important role in the changes of gallbladder function. Experiments have verified the effect of these hormones on the extra - hepatic bile duct.It has been shown that the disturbance of the dynamics of extra - hepatic bile duct play an important role in the formation of gallstones. Mekel and Helm-basch stated that bile stasis played an important role in the pathogenesis of gallstones in 1856. Motilin and substance P may affect the formation of gallstones through the abnormal regulation of dynamics of extra - hepatic bile duct. Now, it is still controversial about the change of plasma motilin in gallstone patients and animals. This study showed that the content of plasma and gallbladder tissue motilin increased significantly in gallstone patients indicating the disturbance of motilin and other gastrointestinal hormones induced the disturbance of bile duct motility and facilitated the formation of gallstones. The difference of species may justify the controversy among the researches. The stimulation of stones to the vagus nerve of the gallbladder wall may increase the release of motilin. Moreover, the metabolism disturbance of motilin can lead to the reduction of degradation of motilin. Large dosage of motilin may probably lead to the contraction of Oddi's sphincter in animals,inhibit the excretion of bile juice from the bile duct to the duodenum, increase the pressure of bile duct and cause the stasis of bile juice. Furthermore, the contraction of Oddi s sphincter can also excite the vagus nerve, promote the secretion and release of the hormones that can inhibit the niotility of gallbladder. Further studies are needed to testify this explanation. |
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