A Cross-sectional Study On Postmenopausal Metabolic Associated Fatty Liver Disease And The Mechanism Of Yanggan Jieyu Granule

Objective:To collect data on metabolic associated fatty liver disease(MAFLD)in postmenopausal women through cross-sectional study,analyze the pathogenic characteristics,risk factors,traditional Chinese medicine constitution,and syndrome patterns of MAFLD in postmenopausal women,establish a risk prediction model,and visualize the model with a nomogram;and further analyze the mechanism of Yanggan Jieyu Granule in treating rats in a MAFLD model through pharmacodynamics and transcriptomics,so as to realize in-depth exploration of traditional Chinese medicine in treating postmenopausal MAFLD.Methods:1.This study retrospectively collected the baseline data,physical examination results,medical history,metabolic associated risk factors,diet,traditional Chinese medicine constitution,syndrome types and other related information on postmenopausal women from January 2023 to October 2023 to study the pathogenic rule of this disease.2.Further univariate logistic regression and stepwise regression were performed on the collected data to screen significant indicators independently related to MAFLD,and to establish a nomogram model for predicting MAFLD.The performance of the nomogram was quantified using C-index and calibration curve,and the model was evaluated using Area Under Curve(AUC)and Decision Curve Analysis(DCA).3.A postmenopausal MAFLD female rat model was established and the modeled rats were divided into different groups to receive intervention with Yanggan Jieyu Granule.The effect of Yanggan Jieyu Granule on postmenopausal MAFLD female rats was observed by detecting serum AST,AST,TC,TG,liver coefficient,and pathological examination in rats.The mechanism of Yanggan Jieyu Granule on treating postmenopausal MAFLD was elucidated by using spatial transcriptomic analysis,differential analysis,enrichment analysis,KEGG analysis,and Western blot(WB).Results1.This study included 942 eligible postmenopausal women with an average age of 59.68 years,among which 313 women(33.23%)were diagnosed with MAFLD.The postmenopausal population showed more significantly insufficient intake of vegetables,fruits,and dairy products,accounting for 86%,72%,74.31%,and 92.36%,respectively;and there was a general insufficient exercise among the population,with only 1.17% of the population achieving aerobic exercise at moderate intensity or above for >150 minutes per week.The postmenopausal MAFLD population also showed higher incidence of overweight,hypertension,hyperlipidemia,diabetes and hyperuricemia.Furthermore,Qi-stagnation(25.48%),Yang-deficiency(17.52%),damp-heat(14.65%),and Yin-deficiency(11.36%)constitutions were common in postmenopausal women;while damp-heat(19.81%),Qi-stagnation(18.85%),and phlegmdampness(14.38%)were common in postmenopausal MAFLD population.Among them,subjects with phlegm-dampness(48.91%),damp-heat(44.93%)and blood-stasis(40.30%)constitutions had relatively higher incidence of MAFLD.Of the 313 postmenopausal MAFLD patients diagnosed in this study,there were 190 cases(60.70%)of liver depression and spleen deficiency syndrome,26 cases(8.31%)of phlegm obstruction syndrome,39 cases(12.46%)of damp-heat brewing and binding syndrome,13 cases(4.15%)of phlegm and blood stasis syndrome,17 cases(5.43%)of liver-kidney Yin-deficiency syndrome,and 28 cases(8.95%)of spleen-kidney Yang-deficiency syndrome.2.Multivariate logistic regression analysis showed that occupation(worker),BMI,waistto-hip ratio,number of abortions,anxiety,hypertension,hyperlipidemia,diabetes,hyperuricemia,diet(meat,processed meat)were risk factors for MAFLD in postmenopausal women.A risk prediction model was established accordingly.The C index of the nomogram was 0.827(95% CI 0.794-0.860)and 0.787(95% CI 0.728-0.846)in the prediction group and the validation group,respectively.The calibration curves 1 and 2 of the nomogram were close to the diagonal.The AUC of the prediction group was 0.827,with a sensitivity of 0.784 and a specificity of 0.735;while the AUC of the validation group was 0.787,with a sensitivity of0.674 and a specificity of 0.772.The DCA curve revealed that the nomogram exhibited more preferable net benefit in predicting MAFLD in postmenopausal women.3.Compared with the model group,Yanggan Jieyu Granule group had improved liver index(P<0.01),ALT(P<0.01),AST(P<0.001),TC(P<0.05)and TG(P<0.01);and HE staining suggested alleviated infiltration of inflammatory cells in the portal area and interstitial tissues.Further WB results suggested that this treatment could up-regulate Ca M(P<0.001),ERα(P<0.05)and phosphorylated AMPK(P<0.01),but inhibit ACC2(P<0.05).Conclusions:1.The cross-sectional study revealed that qi-stagnation constitution,yang-deficiency constitution,damp-heat constitution,and yin-deficiency constitution were prevalent constitutional types among postmenopausal women.In the postmenopausal MAFLD group,the common constitutional types were damp-heat constitution,qi-stagnation constitution,and yindeficiency constitution.Further analysis based on card type in the postmenopausal MAFLD group indicated that qi depression,qi deficiency,internal dampness and heat were the most frequent disease factors associated with spleen and liver dysfunction.It can be observed that liver stagnation and spleen deficiency along with internal dampness and heat are commonly witnessed in postmenopausal MAFLD.2.The multivariate logistic regression analysis identified 11 independent risk factors for postmenopausal women,including occupation(worker),BMI,waist-to-hip ratio,number of abortions,anxiety,hypertension,hyperlipidemia,diabetes,hyperuricemia,meat frequency and processed meat diet habit.Based on these findings,a nomogram model was developed and validated to predict the risk of MAFLD in postmenopausal women.This model can be utilized for early screening of MAFLD in this population.3.In this study,a multivariate logistic regression was conducted to screen 11 independent risk factors for the specific group of postmenopausal women.Subsequently,a nomogram model was developed and validated to predict the risk of MAFLD.This model holds potential for early screening of MAFLD in postmenopausal women and bears significant implications for disease prevention and treatment.However,it is important to note that certain limitations may arise due to population selection,necessitating verification through multi-center studies with larger sample sizes.