| Objective: The liver-spleen correlation theory,first appeared in the Neijing and Naijing period,has tended to form a complete ideological system under the constantly supplement and perfection of doctors in successive dynasties.On the basis of modern meidicine,liver-spleen correlation theory has been more profoundly interpreted and developed,and is widely applied to the analysis of etiology and pathogenesis,the judgment to diagnosis and prognosis,and the guidance on the prevention and recuperation.Primary biliary cholangitis(PBC)is a kind of autoimmune and chronic progressive liver disease.Stagnation of liver qi and spleen deficiency is the basic syndrome according to the clinical differentiation.Tiaogan Lipi decotion(TGLP),the exerperience decotion self-designed by my tutor Guo Xiaoxia in the guidance of liver-spleen correlation theory,has been found to play a significant role in improving the liver and intestinal function,and the PBC patient’s overall quality of life after the validation of clinical efficacy for many years and the result of preliminary experimental study.This research aim to explore and summarize the connotation and application of the liver-spleen correlation theory and achieve mastery through a comprehensive study on its ancient and modern thoughts in order to make better use of it in the area of disease’s syndrome differentiation and classification and clinical treatment.We also propose to establish PBC animal model by intraperitoneal injection of Polyinosinic-polycytidylic acid(poly I: C).Studying the related genes’ expressions of inflammatory cytokine TNF-α and TLR4/NF-κB signaling pathway in the tissue of terminal ileum of PBC model mice,we want to discuss the action mechanism of TGLP combined with ursodesoxycholic acid in the intervention to PBC model mice and provide experimental basis for exploring the new targets for the prevention and control of PBC.Method:1.Theoretical researchThe retrieval methods of collecting literature and referencing books are used in summing up the essence and connotation of liver-spleen correlation theory from the angle of traditional Chinese medicine and modern medicine.And then we interpreted the development and application of this theory.2.Experimental research100 C57BL/6 female mice were randomly divided into the normal group,the model group,the positive group(UDCA 0.1g·kg-1·d-1),the Chinese medicine group(TGLP5.4g·kg-1·d-1),the positive+Chinese medicine group(TGLP 5.4g·kg-1·d-1+UDCA 0.1g·kg-1·d-1),20 in each group.Poly I:C 5mg/kg was used to copy PBC mice model.All mice,fasted for 24 h before processing,were executed by cervical dislocation method at week 8 and week 16.Collect blood,livers,terminal ileum in mice for testing.(1)Positive expression rates of anti-mitochondrial antibodies subtype M2(AMA-M2)in mice serum were detected by Enzyme-linked immunosorbent(ELISA).The biochemical criterions(ALT,AST,ALP)in mice serum were analysed through the automatic biochemical analyzer.According to the results of the biochemical criterions,choose three specimens in each group to observe pathological changes in liver tissues.(2)Also,pathological changes in terminal ileum tissues were observed with HE staining method.(3)The expression levels of TNF-α in terminal ileum tissues were detected by the method of ELISA and quantitative real-time PCR(q RT-PCR).The relative expression levels of TLR4 and NF-κB m RNA in the terminal ileum tissues were detected by the method of q RT-PCR.Result:1.Theoretical ResearchGut function of modern medicine belongs to spleen function of traditional Chinese medicine.Liver-spleen correlation can be characterized by liver-gut interrelation.The scholars in later ages have done much to interpret it based on the nerve-endocrine system and intestinal mucosa barrier.They think that gut-liver axis theory is a complement to the liver-spleen correlation theory.2.Experimental study(1)According to the testing results of biochemical criterions in mice serum,the levels of ALT,AST,ALP in the PBC model group have increased compared to the normal group on week8 and week 16,and the difference is statistically significant(P<0.05).(2)According to the testing results of AMA-M2 in mice serum,the positive rates of AMA-M2 in the model group were respectively 56% and 78%.However,AMA-M2 testing results in the normal group were negative.(3)HE staining results of mice liver tissues show that,with the extension of model-building time,a large number of inflammatory cells,even forming into epithelioid granulomas,infiltrate at expanded portal areas in the model group;Epithelial cells of interlobular bile ducts become degeneration and damaged;Around the portal areas,hyperplasia of small bile ducts accompany with infiltration of lymphocytes.Fragmental necrosis appear at hepatic limiting plates.There are not obvious inflammatory cell infiltration at and around portal areas in the nomal group;and the structures of hepatic lobules can be clearly seen.Masson staining results show that,with the extension of model-building time,a great number of blue collagen fibers,abnormally distributing at expanded and edematous portal areas in the model group,form a wide fiber interval,even accompanied with bridging fibrosis individually.The false flocculus can not been seen under the microscopic.The lumens of small bile ducts become expansile and twisty.As to the normal group mice,the architecture of hepatic lobules is nomal and blue-green collagen fibers do not distribute obviously at portal areas.(4)HE staining results of mice terminal ileum tissues revel that,in the model group,intestinal mucosal villi gradually atrophy,become shorter,and fracture with the model-building time.The gap below intestinal epithelium is broadening.Quantities of inflammatory cells infiltrat lamina propria and even encroach on mucosal muscularis.The varying degrees of separations between the epithelium and lamina propria are visible,accidentally accompanied with capillaries exposure in lamina propria.Destruction and incompleteness can even appear in the lamina propria.Compared with the model group,the pathological changes of the treatment groups were all improved,intestinal mucosa villus becoming neat,mucous membrane layer being relatively close,the gaps under the epithelium narrowing,infiltration of inflammatory cells decreasing significantly.The pathological damages in the positive+Chinese medicine group are obviously improved.Its pathological grading is the lowest and the difference is statistically significant compared to the model group(P<0.05).(5)The TNF-α testing results in the terminal ileum reveal that,the expressions of TNF-α in each group do not change obviously with the model-building time.Compared with the rest of the groups,TNF-α express on a high level in the model group mice;the difference is statistically significant(P<0.05).Compared between the treatment groups,there is nostatistically significant difference(P> 0.05).(6)The TLR4 and NF-κB m RNA testing results in mice terminal ileum reveal that,the expression level of TLR4 m RNA is higher in the model group at each temporal points compared with the normal group;but there was no statistically significant difference(P>0.05).Compared with the positive+Chinese medicine group,the difference was statistically significant(P<0.05).Compared between the treatment groups,there was no statistically significant difference(P>0.05).The expression level of NF-κB m RNA is higher in the model group at each temporal points.Compared with the other groups,the difference was statistically significant(P<0.05).In all treatment groups,the difference between the positive+Chinese medicine group and Chinese medicine group is statistically significant(P<0.05).Conclusion:1.Gut-liver axis theory is a complement to the liver-spleen correlation theory.The development and perfection of the liver-spleen correlation theory play an important role in clinical guidelines.2.The using of poly I:C could successfully clone early animal model of primary biliary cholangitis.3.TGLP combined with UDCA has a protective effect to intestinal mucosal barrier in PBC model mice.The action mechanism may be closely related to negatively regulate TLR4/NF-κB signaling pathway and then reduce the expression of proinflammatory cytokines TNF-α. |
PDF Full Text Request