Study On The Effect Of Qing’e Pills In Anti-Osteoporosis From The Perspective Of Intestinal Flora

Research background and purposeQing’e pills,first published in the Song Dynasty’s Taiping Benefit the People and Drug Administration,is usually used to invigorate the kidney and strengthen bones.It has been used for the prevention and treatment of clinical osteoporosis since the Tang Dynasty.The research group has used a rat ovariectomized model to measure a series of indicators in the early stage to prove that Qing’e Pills is an effective prescription for treating osteoporosis.Qing’e Pills can regulate the balance of bone metabolism,promote bone calcium absorption,regulate calcium and phosphorus balance,inhibit osteoclast activity,stabilize bone metabolism,and promote osteoblasts,generate more bone matrix,and restore osteoporosis in rats Normal bone metabolism.The research team co-incubated the active ingredients in Qing’e Pills with the intestinal bacteria population of rats.The main metabolites are enterodiol,intestinal fat,psoralen and isopsoralen and other phytoestrogens.Indicating that the intestinal flora converts the prototype active ingredients into active phytoestrogens to a certain extent.By comparing the effects of the active ingredient prototype of Qing’e Pills and related metabolites on osteoblasts,it was found that psoralen and psoralen chalcone have significant proliferation effects on osteoblasts,and psoralen can increase alkalinity.Sex phosphatase activity enhances the differentiation of osteoblasts.It shows that the biotransformation of intestinal flora to Qing’e Pills is beneficial to improve the bone repair ability of salt-baked Qing’e Pills for osteoporosis.Therefore,identifying the effective and specific mechanism of Qing’e Pills in the treatment of osteoporosis has important guiding significance for the clinical use of Qing’e Pills.Further literature research found that the relationship between intestinal microecology and bone metabolism has become a hot spot in the field of medical research.Injecting normal mice with drugs that inhibit estrogen(imitating menopausal women)has significant bone loss,and injecting the same drugs into sterile mice has no significant bone loss.Therefore,the intestinal flora is related to the occurrence and development of osteoporosis.People gradually realized the importance of maintaining intestinal microecological balance in the process of treating diseases and began to turn their attention to traditional Chinese medicine.Chinese medicine is rich in chemical components.In addition to bioactive components such as flavonoids,terpenes,anthraquinones,alkaloids,and steroids,it also contains a variety of nutrients such as protein and vitamins.A large number of studies have shown that traditional Chinese medicine has a protective effect on the balance of the intestinal microecosystem and can directly or indirectly regulate the imbalance of intestinal flora.This topic aims to clarify the efficacy of Qing’e Pills on osteoporosis,and to explore the possible mode of action of Qing’e Pills from the perspective of intestinal flora,and to provide new ideas and strategies for the research of Chinese medicine.Research contents and resultsIn this study,SD rats were selected as the research object,and the osteoporosis model of bilateral ovariectomized rats was used to evaluate the effect of salt burning on the efficacy of Qing’e Pills in the treatment of osteoporosis in rats.The administration was continued 6 weeks after surgery8 week.The therapeutic effects of Qing’e Pill on osteoporosis were evaluated by indicators such as body weight,uterine index,Micro-CT parameters,bone biochemical indicators,and bone biomechanical parameters.Using 16s rRNA technology to measure the content of intestinal flora in each group,it was found that there were significant differences in the abundance and diversity of intestinal flora between the sham operation group and the model group,and the abundance and diversity of intestinal flora remained the same as the model after administration.There is a significant difference between the two groups(P<0.05),which tends to be the sham operation group.Compared with the sham operation group,the differential flora Lactobacillus and Akkermansia of the model group decreased,and the unclassified_f_Lachnospiraceae,Blautia,Christensenellaceae_R-7_group increased,and tended to the normal group level after administration.Through correlation analysis,it is found that Lactobacillus and Akkermansia are positively correlated with BMD,BV/TV,BS/TV and Tb.N,indicating that probiotics such as Lactobacillus are beneficial to the prevention and treatment of osteoporosis.In order to explore the mechanism of this change,plasma non-targeted metabolomics was used to collect and analyze the characteristic maps of relevant biological samples with the help of UPLC-Q/TOF-MS technology.A total of 44 different components and negative ions were detected in the positive ion mode.In the model,a total of 29 different components were detected,and 19 potential biomarkers related to osteoporosis were screened out,involving 11 metabolic pathways,including unsaturated fatty acid biosynthesis pathway,glycerophospholipid metabolism pathway,and sub-Oleic acid metabolism pathway,arachidonic acid metabolism pathway,α-linolenic acid metabolism pathway,primary bile acid biosynthesis pathway,sphingolipid metabolism pathway,steroid hormone biosynthesis pathway,tryptophan metabolism pathway,aminoacyl tRNA biology Synthesis pathway and retinol metabolism pathway.The content of short-chain fatty acids,the metabolites of intestinal bacteria,was analyzed by GC-MS technology.The results showed that compared with the sham operation group,the content of acetic acid and propionic acid in the model group was significantly reduced.Significantly increased.By HE staining the rat ileum,compared with the sham operation group,the ileum of the model group has a large amount of mucosal epithelium shedding,and the glands are deformed and atrophy or the structure is severely lost.The Qing’e Wansheng product group has severely deformed and atrophy.There was a certain degree of improvement in the pill salt product group.ELISA method was used to detect the levels of IL6,IL-17,TNF-α and IFN-γ in the rat serum.Compared with the sham operation group,the serum levels of IL-6,TNF-α and IL-17 in the model group increased significantly.High(P<0.01),there was no significant difference in IFN-y levels(P>0.05).Compared with the model group,there was a difference in serum IL-17 levels in the Qing’e pill raw product group(P<0.05),and the rest had no significant difference(P>0.05);the Qing’e pill salt group had no significant difference in serum IL-6,The levels of TNF-α and IL-17 were significantly reduced(P<0.01),but there was no difference in the levels of IFN-γ(P>0.05).It is speculated that the intestinal flora may be a potential therapeutic target for osteoporosis.Qing’e pills may treat osteoporosis by influencing the intestinal flora to regulate host metabolism,intestinal bacteria’s autonomous metabolism and immune pathways,and finally transplant the fecal flora of the Qing’e pills group to the osteoporosis group.We found that fecal bacteria transplantation can effectively alleviate the inflammation and damage of the intestinal barrier in rats,and at the same time change the host metabolism and short-chain fatty acid content of the rat,verifying that Qing’e Pills may regulate the host metabolism and intestinal intestines by affecting the intestinal flora.Autonomous metabolism of bacteria and immune pathways to treat osteoporosis.Conclusion and SignificanceBased on the above results,we found that Qing’e Pills can change the abundance and composition of the intestinal bacteria in rats,increase the abundance of lactobacilli,affect host metabolism,and increase the production of short-chain fatty acids,reduce inflammation in the body,and thus play a role in osteoporosis.The alleviating effect of the disease.Fecal bacteria transplantation also proved the possibility of intestinal flora for the treatment of osteoporosis.This study clarified the possible mechanism of Qing’e Pills in the treatment of osteoporosis from the perspective of intestinal flora,and provided a new reference for the research thinking of Chinese medicine.