Regulatory Mechanism Of Shengli Decoction On Aquaporin In Submandibular Gland Of Sjogren 's Syndrome Model Mice

Sjogren’s syndrome (Sjogren’s syndrome, SS) is a chronic autoimmune disease, in violation of the lacrimal gland, salivary glands and other exocrine glands resulting in reduced secretion, dry mouth, dry eye symptoms as the initial performance, often accompanied by a variety of organs damage. Some patients may develop permaent organ damage or evenlife-threatening condition. The etiology of SS by modern medicine study may be genetic, viral infections, immune dysfunction and other factors.Traditional Chinese medicine has made some achievements in the clinical treatment of pSS by delaying the disease progression, relieving symptoms, and significantly improving life qulity. Zengye Decoction from "Treatise on Warm Disease" is proved to be effective in treating SS. Preliminary study indicate that Zengye Decoetion can increase the expression of aquaporin AQP1, AQP5 in submandibular gland, promote glandular secretion, relieve dry mouth symptoms of Sjogren’s syndrome model mice. Wherease the expression level of AQP1 and AQP5 in submandibular gland were significantly different, the AQP5 expression level in model group was significant downregulated while AQP1 up-regulated, the Zengye Decoction can continue to increase the AQP1 expression which indicate the regulation of the two aquaporins may be different in signaling pathwayor immunological mechanisms.PURPOSE:This study focus on:1 explore the pharmacological effects of improving the phathological changes of mandibular salivary gland and regulating AQP1,AQP5 expression of Zengye Decoction on pSS mice model.2 clarify whether Zengye Decoetion regulate the expression of AQP1/AQP5 through the 3 signalling pathways TNF-a/TNFR/IKK/NF-κB, TRAF6/JNK/c-Jun, MKK7p38 lightning/c-Fos, to interpret the treating method of "increasing the water essence to norish body tissue" in traditional Chinese medicine.METHODS:Thirty BALb/C mice, were randomly divided into 3 groups:control group, model group, add Zengye decoction group, mice gland antigen preparation was performed by differential centrifugation method. Multipoint subcutaneous injection of freund’s complete adjuvant of submandibular gland immune antigen was performed in Model group and Zengye decoction group for 21 days.Animal weight and food intake amount was recored through the whole experiment days. Mice submandibular gland index is calculated after animals were sacrificed, submandibular gland pathological section were stained with H-E for histologic observation by optical microscope.Protein expression of AQP1 and AQP5 of submandibular gland in pSS mice model were measured with Immunohistochemical techniques.Westerning bloting method was appled to detect NF-κB, c-Jun and p38 expression level in experimental animal.RESULTS:Compared with the control group, the submandibular gland index of pSS mice model was significantly decreased (P< 0.01); glands atrophy was dectected in submandibular gland tissue by histologic observation, focal lymphocytic infiltration was observered, which indicate typical pathological changes in sjogren’s syndrome. AQP1 and AQP5 expression of Submandibular glandin Zengye decoction group was increased significantly, while in mice modle group, AQP1 expression was increased, AQP5 expression was declined. Westernblot results showed positive expression NF-κB, c-Jun, p38,which indicating the possible regulating pathway of Aquaporin in mice model, were significantly higher than that of control group (P< 0.01), Zengye decoction can significantly restrain the expression of NF-κB, c-Jun, p38 compared with model mice (P< 0.01).CONCLUSION:This sduty showed that Zengye decoction can significantly increase submandibular gland index in SS mice, inhibiate lymphocyte infiltration in submandibular gland, prevent pathological process of submandibular gland atrophy and fibrosis, upregulating the expression of AQPs and influence its distribution, promotingwater secretion and transportation in saliva gland cells. Further study indicate that three signaling pathways TNF-α/TNFR/IKK/NF-κB, TRAF6/JNK/c-Jun, MKK/p38 lightning/c-Fos thatmight be invoved in the regulation of aquaporin in the pSS mice model;Zengye decoction can significantly decrease the expression level of NF-κB, c-Jun, p38, which might lead to the conculsion that Zengye decoction can regulate the expression of aquaporin via TNF-a/TNFR/IKK/NF-κB, TRAF6/JNK/c-Jun, MKK/p38 lightning/c-Fos pathways.