| ObjectiveTo build up the animal model of submandibular gland injury radiation-induced as well as to test the damage of submandibular gland,we adopted the technology of HE staining,PCR to detect gland' s change in morphology and the expression of AQP1 and AQP5-after all interventions were done within 28 days.Additionally,the experiment explored the effect and the mechanism of plum spray to submandibular gland injury radiation-induced at molecular pathology level,which lays the foundation for clinical application,easing the burden on clinical nursing work,increasing patients'comfort,and reducing the economic burden of patients.MethodsMethod of experiment one:115 Wistar rats at 6?7 weeks and weighs about 180?220 grams,were randomly divided into irradiated group(IG,n=59)and sham-irradiated control group(SIG,n=56)and the experiment has been approved by ethical review board.IG was given single dose of 18Gy irradiation located on the submandibular glands,while SIG didn't exposure to radiation.The two group were divided into eight point times after radiation such as the 1th,3tn,7th,14th,21th,28th,35th,42th day,after that their dietary intake,volume of saliva and other indicators were examined.Each group of each time point took 7-8 rats to dissect the submandibular gland tissue for histopathological examinations.Morphology examinations were assessed by HE staining and the expression of AQP1 and AQP5 was assessed by PCR technology.Through the volume of water consumption and saliva,morphology examination,expression of water channel AQP1/AQP5,we determined whether the rats model of submandibular gland injury radiation-induced was built successfully or not.Method of experiment two:As a randomized controlled trail,175 Wistar rats were randomly divided into five groups,including sham-irradiated control group(SIG,n=35),irradiated group(IG,n=35),normal saline group(NSG,n=35),pilocarpine group(PG,n=35)and dark plum spray group(DPSG,n=35).Except SIG,other four groups were given one-time 18Gy irradiation locally on the submandibular glands,after that,we collected the volume of saliva and submandibular gland on the 1th,3th,7th,14th,28th day.With HE staining and PCR technique,we detected changes in morphology and the function of the salivary glands and the expression of water channel protein AQP1 and AQP5.ResultsResult of experiment one:In 1?21 days after irradiated,there was statistically significant higher water consumption(6.42±1.91)ml in IG compared with SIG(6.42±1.91ml vs 4.82±1.20 ml,P<0.05).During 42days after radiation,the saliva secretion of IG was significantly lower than SIG on the 7th,21th,28th,42th day(P<0.05).There were significantly differences in the comparison of gland radio among two groups on the 1th,21th,28th,35th,42th day(P<0.05).Furthermore,compared to SIG,the expression in IG of Water channel protein AQP1 and AQP5 was significantly suppressed and HE pathology revealed that the submandibular glands of rats became atrophy,inflammatory and reduced cytoplasm secretion after irradiation and it was most serious on the 42th day after radiation.Result of experiment two:There was statistically significant lower saliva flow rate in IG compared with SIG on the 7th,28th day(P<0.05).Moreover,the saliva flow rate of DPSG was statistically significant higher than IG and SIG in 7 days(P<0.05),while Saliva composition had no obviously changed after intervened by dark plum spray.The submandibular gland cells in DPSG was less atrophy,inflammatory and more cytoplasm secretion than the other irradiated groups and the restoration of submandibular gland cells were better on the 14th,28th day.In addition,PG and DPSG could promote the gene and protein expression of water channel of AQP1 and AQP5(P<0.05).ConclusionAfter given single dose of 18Gy,Animal model of submandibular gland injury radiation-induced was established successfully.The mechanism we find is that when submandibular gland cells were damaged by radiation,the expression of water channel of AQP1 and AQP5 were inhibited at the same time,and then water transmembrane was damaged,along with saliva secretion declined and gland damage became increasingly seriously.Furthermore,after 2 weeks intervention,dark plum spray promotes water channel of AQP1 and AQP5 high expression to increase the production of saliva,and to repair the damage of submandibular gland.But there is no correction in saliva composition changes after intervened by dark plum spray. |
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