| PrefaceDiabetic gastroparesis (DGP) is one of the common complications of diabetes. At present, it is considered to be caused by functional disturbance of gastric intestinal smooth muscle neuromuscular junction as a result of long-term diabetes process, so abnormal intestinal peristalsis causes nausea, vomiting, abdominal discomfort, abdominal distension and so on. DGP seriously influences the quality of the patients' lives and is difficult to be treated. Development of the molecular biology and cytology makes it possible to deeply study evolutionary process of DGP in molecular level as well as cell biological effect.It is conformed that MTL, GAS, SS, VIP and SP play important role in regulating gastrointestinal movements. These gut hormones also exist in central nervous system, so they are called Brain gut peptide, Some of which are neurotransmitter or neuromodulator, then gastro intestinal system and nervous system are connected together like this. Someresearch have indicated that the Brain gut peptide which are distributed widely in central nervous system, peripheral nervous system , gastrointestinal tract and many other organs belong to G protein(GTP-binding protein) that regulates gastrointestinal motility and contracts or relaxes intestinal smooth muscle. Their biological action is wide, and it has been conformed that MTL > VH\ SP correlate with DGP closely. The role GAS and SS as well as their receptors play in DGP has not been reported till now.GAS secreted by G cells exists in gastrointestinal tract especially in gastric antrum mucosa. Its main function is stimulating gastric acid secretion, nutritional supporting of mucosa from acid area and stimulating movement of gastric antrum. The GAS level in serum is often high in the diabetics who have autonomic neuropathy. SS can inhibit GAS intensively, so when hyperglycemia appears, secretion of SS decreases and inhibits GAS weakly and resulting in GAS level's increase in serum. The process GAS stimulates incretion of gastric acid is mediated by GAS/CCK-B receptor. SS is a ring type polypeptide which has two main kinds of exist form in mammals, they are 14 and 28 peptides. Besides regulating secretion, SS can regulate multiple physiological activities and takes part in some pathologic process as a neurotransmitter. SS secreted by D cells of gastric antrum can inhibit secretion of multiple gastrointestinal hormones and inhibits gastrointestinal tract movementand absorption of glucose^ triglyceride and electrolytes, so serum glucose transports to tissues more and the blood sugar level degrades.The HP infection rate in DGP patients is 78.61%, which is far more than in patients without DGP(43.85%). The HP infection rate of diabetics without DGP is close to the rate of common healthy crowd(46.5%), which shows that HP infection is related with DGP.This study aims at revealing the pathogenic mechanism of DGP by multiplicity and conduces to clinical treatment and control of diabetes complications.ObjectiveTo explore the effect of GAS> SS and HP in the development of DGP, and the morphous , quantity , distribution and configuration of the secretory cell of GAS (G cell) and the secretory cell of SS (D cell) in gastromucosa tissue, observe their interrelationship and clinical diagnosic significance in DGP.MethodUtilize radioimmunity device and RT-PCR to detect the level and protein express of serum GAS> SS in normal control(17cases), simple delayed gastric emptying group(30 cases), diabetes mellitus with normal gastric emptying group(23 cases), and DGP group(33 cases);apply hybridization in situ method to detect the level of GAS^ SS in the sinusventriculi tissue, to detect the gene sequence of mRNA in every group, and observe the quantity ,morphous and distributionand of G cell and D cell in sinus ventriculi in these groups. Use tachy-urease experimentation and Giemsa staining method to check HP in mucosa of sinus ventriculi in every group cases. Use radioimmunity Methods to determine the contents of serum GAS^ SS in every group cases o Use statistics multiplicity to analysis dependability among GAS n SS and HP as well as the expression of mRNA gene and protein. Approach their interaction in DGP development.ResultsThe radioimmunity device -, hybridization in situ method and RT-PCR yield the tendency coincident results, which hint the indeed biology effect of GAS> SS and HP in DGP morbility and also has found the dependability among the quantity , morphous and distribution of G cell and D cell with GAS> SS and HP in DGP morbility . The detective results of GAS> SS:In the normal control ( I ) : serum GAS is 83.47±14.35pg/ml, serum SS is52.08±15.11pg/ml.2> simple diabetes mellitus group (II ) : The serum GAS is 102.21 + 21.54pg/ml;The serum SS is 42.57 ± 13.56pg/ml, there is obvious difference between the two groups (P<0.05) respectively. 3> The simple gastroparesis group (III) : serum GAS is 98.31 +23.81pg/ml, serum SS is 60.86 ± 14.15pg/ml, There is obviouslydifference compare with the normal control (P<0.05) .^ DGP group (IV) : the serum GAS is 146.58 ±28.83pg/ml, the serumSS is 16.15 + 3.85pg/ml ,there is significant differences compare withnormal control (P<0.01);and there is obvious differences compare withII and III (P<0.05) .The detective results of mRNAHIS in G cell and D cell:1> normal control ( I ) : the positive rate of GAS is 17.65%;the positive rate of SS is 76.47%.2> simple diabetes mellitus group ( II ) : the positive rate of GAS is 26.67%;the positive rate of SS is 56.67%, there is obviously difference between the two groups (P<0.05) respectively.3> The simple gastroparesis group (III) : the positive rate of GAS is 34.78%, the positive rate of SS is 56.52%, there is obviously difference compare with the normal control (P<0.05 ) .4> DGP group (IV ) : the positive rate of GAS is 78.79%, the positive rate of SS is 24.24%, there is obviously difference compare with the normal control (P<0.01);and there is obvious differences compare with II and III (P<0.05) .The PCR detective results of GAS^ SS :U Normal control ( I ) : the I value of GAS is 0.338 + 0.313;the I value of SS is 0.756 + 0.302.2> Simple diabetes mellitus group ( II ) : the I value of GAS is 0.408 ±0.439ps;the I value of SS is 0.521 + 0.232, there is obviously difference between the two groups (P<0.05) respectively. 3 >> The simple gastroparesis group (III) : the I value of GAS is 0.432 ±0.299, the I value of SS is 0.528 + 0.423, there is obviously difference compare with the normal control (P<0.05) . 4> DGP group (IV ) : the I value of GAS is 0.662 + 0.301, the I value of SS is 0.326+0.282, there is obviously difference compare with the normal control (P<0.01);and there is obvious differences compare with II and III (P<0.05) . The electron microscope detective results of G cell and D cell:Ultrastructure of G cell and D cell in sinus ventriculi of simple diabetes mellitus and simple gastroparesis have not obvious changed when compared with normal control group . The G cell in sinus ventriculi of DGP is expressed by that, the change of internal structure is not obviously, the secretory granules are not reduced, beside the number has increased. In D cell of DGP , rough endoplasmic reticulum(RER) distend to capsular, the ribose granula disappears and the cell organelle becomes decrease.The HP of sinus ventriculi by tachy-urease experimentation and Giemsa staining method:There is obviously difference between every group whenexaminated by tachy-urease experimentation and Giemsa staining method (P>0.05). To utilize tachy-urease experimentation and Giemsa staining method, the HP detection rate of normal control simple gastroparesis group is, the simple diabetes mellitus group is, DGP group is (47.1%, 41.2%), (73.3%, 66.7% X (47.8%, 43.5% )> (87.8%, 81.9%) respectively,there is no difference between the two methods (P>0.05) . There is no difference between the simple diabetes mellitus group and the normal control group (P>0.05);there is obviously difference in simple gastroparesis group compared with normal control(P<0.05). There is obviously difference in DGP group when compare with the simple diabetes mellitus group and the normal control group (P<0.001), and obviously difference compare with simple gastroparesis group (P<0.01).All these express of the gastroparesis have relationship with HP infection. Nearly 90% of DGP patients have HP infection, which show that HP infection may aggravate the DGP.HP (+) group: the GAS of DGP group is higher than normal control (P<0.001), and the SS is lower than normal control (P<0.001);HP(-) group: In every group, GAS and SS have no obviously difference (P>0.05).DGP group: the serum GAS of HP(+) is higher than HP(-), and the difference is also obviously (P<0.05);the serum SS of HP(+) is lower than HP(-), and the difference is ALSOobviously (P<0.05).ConclusionsThe increase of GAS and the depression of SS are the factors in the arises of DGP. In D cell of DGP , rough endoplasmic reticulum(RER) distend to capsular, the ribose granula disappears and the cell organelle becomes decrease, those are the main factors which cause the depression of SS and the increase of GAS. HP infection can facilitate the arise of DGP, and has dependability with the secretion of GAS^ SS. |
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