The Molecular Mechanism Of Gan Zao Granule On Sj(o|:)gren Syndrome Mice’s Salivary Gland

Objective:Treated the SS rats with Gan Zao granule, then examine the expression of AQP5, ICAM-1, VCAM-1in submandibular salivary glands and the change of Ca2+in salivary gland primary cells, explore the mechanisms of Gan Zao granule on Sjogren syndrome, offer support to the sputum pent up characteristic of SS.Methods:Submandibular gland tissue was extracted from female Lewis rats, after grinded into homogenate, the supernatant was blended and emulsification with complete Freund’s adjuvant (CFA). The homology rats were injected by the immunogen. After that, subcutaneous injection of DTP vaccine was conducted on the back to enhance immunity. The effects of Gan Zao Granule on rat isolated salivary glands were investigated, the method used was Chinese herb perfusion. The expression of AQP5, ICAM-1, VCAM-1and E-selectin in submandibular salivary glands of rat with SS was shown by Western Blot, The expression of ICAM-1, VCAM-1, E-selectin in serum was tested by ELISA. The concentration of Ca2+in salivary gland primary cells was measured by Fluorescent calcium ion measurement technology.Results:1. According to the perfusion on rat isolated salivary glands, the Gan Zao granule and Dan Di Qiong Yu granule all can increase the secretion of the salivary glands significantly.2. Compared to the control group, the salivary flow rate of SS rats is decreased, the water intake is increased, the model is successful.3. The salivary flow rate, index of salivary gland of SS rats which treated by middle and high does of Gan Zao granule is higher than that treated by low does.4. Western blot method displays:the model group than normal group submandibular gland AQP5expression was significantly reduced, the treatment group compared with the model group was significantly enhanced compared with AQP5; SS rat submandibular gland ICAM-1, VCAM-1, E-selectin expression in model group was significantly increased compared with the normal group, treatment group ICAM-1, VCAM-1, E-selectin expression compared with the model group significantly decreased, indicating Gan Zao granule can reduce the expression of inflammatory cytokines particles, partly alleviate the condition.5. Gan Zao granule high dose group SS rat submandibular gland cells can be detected Ca2+concentration is lower than the model control group, the difference was significant, P <0.01.6. Compared to the control group, the expression of ICAM-1, VCAM-1in salivary gland of SS rats treated by middle and high does of Gan Zao granule decrease, also E-Selectin exhibit a low level.Conclusion:According to the results above, we come to the conclusion that:Gan Zao granule can treat the SS rats, the mechanism is up-regulate the expression of AQP5and decreasing the intracellular concentration of Ca2+, by inhibiting excessive enhanced immune response, reduced lymphocyte infiltration in the submandibular gland, relieve inflammation factors invasion, reduce rat submandibular gland ICAM-1, VCAM-1, E-selectin expression, thus improving salivary secretion, increase saliva flow, relieve dry mouth symptoms of Sjogren syndrome.Significance:By doing Chinese Traditional Herb experiments, explain the mechanism from the molecular level, provide a new idea for treat SS by using the Chinese Traditional Medicine.