1.Improving the Certainty of Evidence in Animal Experiment Systematic Review/Meta-Analysis: An Empirical Study of the GRADE Method
Tengfei LI ; Qingyong ZHENG ; Jianguo XU ; Yiyi LI ; Yongjia ZHOU ; Caihua XU ; Mingyue ZHANG ; Jiexiang TIAN ; Gang WANG ; Jinhui TIAN
Laboratory Animal and Comparative Medicine 2025;45(1):101-111
Animal experiments are essential tools in biomedical research, serving as a bridge between basic research and clinical trials. Systematic reviews and meta-analyses (SRs/MAs) of animal experiments are crucial methods for integrating evidence from animal experiment, which can facilitate the translation of findings into clinical research, reduce translational risks, and promote resource integration in basic research. With the continuous development of the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) methodology, its application in SRs/MAs of animal experiments has gained increasing attention. This article first outlines the principles and specific applications of the GRADE methodology in SRs/MAs of animal experiments, including qualitative descriptive systematic reviews, meta-analyses, and network meta-analyses. It then deeply analyzes the misuse of the GRADE methodology in practice, including incorrect evidence grading, improper classification of evidence, misapplication in qualitative systematic reviews, inconsistencies between the documentation of the upgrading and downgrading process and results, and inappropriate use for making recommendations. Furthermore, this article comprehensively discusses the factors influencing the grading of evidence certainty in SRs/MAs of animal experiments, including the impact of bias risk, indirectness, inconsistency, imprecision, and publication bias on evidence downgrading, as well as the role of large effect sizes and cross-species consistency in evidence upgrading. Finally, in response to the issues discussed, improvement strategies are proposed, including further research and optimization of the GRADE methodology for SRs/MAs of animal experiments, the development of reporting guidelines tailored to the characteristics of SRs/MAs in animal experiment research, and enhanced professional training for researchers in the GRADE methodology. This article aims to improve the quality of evidence in SRs/MAs of animal experiments, strengthen their reliability in clinical decision-making, and promote the more efficient translation of findings from animal experiment research into clinical practice.
2.Effects of loganin on inflammatory response and intestinal barrier damage in septic rats
Can WANG ; Yantao LI ; Zheng ZHOU ; Lupeng WANG ; Yuanyuan GAO ; Shaoxi FAN
China Pharmacy 2025;36(5):574-578
OBJECTIVE To investigate the effects of loganin on inflammatory response and intestinal barrier damage in septic rats by regulating the Ras homolog gene family member A (RhoA)/Rho-associated coiled-coil forming protein kinase 1 (ROCK1) signaling pathway. METHODS A sepsis rat model was established by cecal ligation and puncture, and randomly divided into sepsis group, loganin low-dose group (50 mg/kg loganin, gavage), loganin high-dose group (200 mg/kg loganin, gavage), positive control group (0.2 mg/kg atorvastatin, intraperitoneal injection), and loganin high-dose + lysophosphatidic acid (LPA) group (200 mg/kg loganin gavage and intraperitoneal injection of 10 mg/kg RohA activator LPA). An additional sham surgery group was established. Each group consisted of 10 rats, and medications were administered once every 6 hours for 4 times. After 24 hours of the last intervention, the levels of serum inflammatory factors interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and IL-1β were detected. The pathological changes of ileal tissue were observed and Chiu’s intestinal mucosal injury score was also performed. The levels of intestinal function-lactate dehydrogenase (D-lactate), D-amino acid oxidase (DAO) and endotoxin, the percentages of zonula occludens-1 protein (ZO-1) and Occludin positive staining area, as well as protein expressions of RhoA, and ROCK1 were all detected. com RESULTS Compared with the sepsis group, the percentages of ZO-1 and Occludin positive areas increased significantly in loganin low-dose and high-dose groups; while the levels of IL-6, TNF-α, IL-1β, DAO, D-lactate and endotoxin, Chiu’s intestinal mucosal injury score as well as protein expressions of RhoA and ROCK1 decreased significantly (P<0.05); the destruction of rat ileal tissue was alleviated, and tissue edema and inflammatory infiltration were significantly reduced; moreover, the improvement effect in loganin high-dose group was superior to that in loganin low-dose group (P<0.05). Compared with loganin high-dose group, RhoA activator LPA reversed the trend of changes in the above indicators (P<0.05). CONCLUSIONS Loganin can alleviate inflammatory response and intestinal barrier damage in septic rats, the mechanism of which may be associated with inhibiting RhoA/ROCK1 signaling pathway.
3.Effect of deep muscle stimulation combined with electromyographic biofeedback on the spasms of the triceps surae and gait changes after stroke
Qiming ZHANG ; Di LIAO ; Zhiliang ZHONG ; Lihua LIN ; Xiang ZHENG ; Qiong LI ; Sharui SHAN
Chinese Journal of Tissue Engineering Research 2025;29(2):385-392
BACKGROUND:Deep muscle stimulation has the effects of releasing muscle adhesion,relieving muscle spasm,improving and restoring muscle compliance and elasticity.Electromyographic biofeedback therapy can promote nerve recovery and improve lower limb motor function and gait. OBJECTIVE:To observe the effect of the effect of deep muscle stimulation combined with electromyographic biofeedback therapy on the spasm of the triceps surae and gait changes after stroke by using a digital muscle detector and three-dimensional gait analysis system. METHODS:A total of 72 patients who met the inclusion criteria were selected from the Rehabilitation Department of the First Affiliated Hospital of Guangdong Pharmaceutical University from October 2020 to October 2023.And they were enrolled and randomly divided into two groups(n=36 per group):a control group and a combined group.The control group received routine rehabilitation therapies,electromyographic biofeedback and pseudo deep muscle stimulation,while the combined group received true deep muscle stimulation treatment on the basis of the control group,five times per week,for 4 consecutive weeks.The oscillation frequency and dynamic stiffness of the affected gastrocnemius muscle,active range of motion of the ankle dorsiflexion muscle,electromyographic signal of the tibialis anterior muscle,Fugl-Meyer assessment of the lower limbs,and three-dimensional gait analysis parameters were statistically analyzed before and after treatment in two groups. RESULTS AND CONCLUSION:After treatment,oscillation frequency and dynamic stiffness values of the inner and outer sides of the affected gastrocnemius muscle in both groups of patients were significantly reduced compared with before treatment(P<0.05),and the combined group showed a more significant decrease compared with the control group(P<0.05).The active range of motion of the ankle dorsiflexion muscle,electromyographic signal of the tibialis anterior muscle,and Fugl-Meyer scores after treatment were significantly increased or improved compared with before treatment(P<0.05),while the combined group showed a more significant increase or improvement compared with the control group(P<0.05).In terms of gait parameters,the walking speed,frequency,and stride in both groups of patients were significantly increased compared with before treatment(P<0.05),while the combined group showed a more significant increase compared with the control group(P<0.05).The percentage time of support phase on the healthy side was shortened compared with before treatment(P<0.05),while the combined group showed a more significant decrease compared with the control group(P<0.05).In addition,there was no significant difference between the two groups except for the percentage of healthy side support(P>0.05).To conclude,the combination of deep muscle stimulation and electromyographic biofeedback can effectively alleviate triceps spasm in the short term after stroke,improve ankle dorsiflexion function,enhance lower limb motor function,and improve gait.The treatment effect is significant and worthy of clinical promotion and application.
4.Effect of electroacupuncture combined with low-frequency transcranial ultrasound stimulation on the electroencephalographic signals of rats with traumatic brain injury
Simiao GAO ; Xue HAN ; Xiaoguang WU ; Jinyu ZHENG ; Fangwen GAO ; Kuihua LI ; Yong PENG ; Lanxiang LIU
Chinese Journal of Tissue Engineering Research 2025;29(2):402-408
BACKGROUND:Traumatic brain injury is a condition in which the normal function of the brain is disrupted by a bump or impact to the head.It is necessary to find effective treatments and objective targets that can help doctors diagnose the injury status and restore the brain function of patients. OBJECTIVE:To explore the effect of electroacupuncture combined with low-frequency transcranial ultrasound stimulation on the electroencephalographic signals of rats with traumatic brain injury. METHODS:Forty 6-week-old SPF male Sprague-Dawley rats were randomly divided into five groups:sham group,model group,electroacupuncture group,low-frequency transcranial ultrasound stimulation group and combined group(electroacupuncture+low-frequency transcranial ultrasound stimulation),with eight rats in each group.Feeney weight-drop method was used to establish the animal model of traumatic brain injury.In the sham group,the bone window was only opened without impact.Interventions were started at 1 day after modeling.Electroacupuncture in the electroacupuncture group,low-frequency transcranial ultrasound stimulation in the low-frequency transcranial ultrasound stimulation group,and electroacupuncture+low-frequency transcranial ultrasound stimulation in the combined group were performed for days in total.The modified neurological severity scale score for assessing rats'neurological deficits was performed at 8 hours after modeling.The percentage of spontaneous alternation behavior in the Y-maze was measured at 7 days after modeling.Then,the electroencephalographic signals were collected and electroencephalographic data of α,β,θ,and δ waves were extracted by fast Fourier transform,and the value of oscillation amplitude and energy ratio were calculated in α,β,θ,and δ waves,as well as the Lempel-Ziv complexity and sample entropy. RESULTS AND CONCLUSION:Compared with the sham group,the modified neurological severity scale scores in the model group,electroacupuncture group,low-frequency transcranial ultrasound stimulation group and combined group were significantly increased at 8 hours after modeling(P<0.05).Compared with the sham group,the value of oscillation amplitude in δ wave and the value of δ energy ratio were significantly increased in the model group at 7 days after modeling,meanwhile the percentage of spontaneous alternation behavior in Y-maze,and the value of α/β energy ratio,Lempel-Ziv complexity,and sample entropy were significantly decreased(P<0.05).Compared with the model group,the value of oscillation amplitude in α and δ waves was significantly decreased in the combined group(P<0.05),while the value of α/β energy ratio was significantly increased(P<0.05)and the value of δ energy ratio was significantly decreased(P<0.05)in the electroacupuncture group,low-frequency transcranial ultrasound stimulation group and combined group.Compared with the electroacupuncture group and low-frequency transcranial ultrasound stimulation group,the value of δ energy ratio was significantly decreased in the combined group(P<0.05),while the percentage of spontaneous alternation behavior,the value of α/β energy ratio,the Lempel-Ziv complexity,and the sample entropy were significantly increased(P<0.05).To conclude,abnormal electroencephalographic signals can appear in rats with traumatic brain injury,while the electroacupuncture combined with low-frequency transcranial ultrasound stimulation can alleviate the abnormal electroencephalographic signals in rats,which suggests the electroencephalographic frequency domain value and nonlinear features can be used to assess the severity of traumatic brain injury.
5.Treating acute type Ⅲ-Ⅴ acromioclavicular joint dislocation with single tunnel fixation versus tunnel-free suspension fixation of the coracoid process under shoulder arthroscopy
Yongtao ZENG ; Hongcheng ZHENG ; Nacikedaoerji ; Refati·Nijiati ; Li SHU ; Xu LIU ; Hongtao CHEN
Chinese Journal of Tissue Engineering Research 2025;29(5):1036-1042
BACKGROUND:At present,there are few reports on the postoperative efficacy of arthroscopic coracoid tunnel-free suspension fixation and coracoid single tunnel fixation in the treatment of acromioclavicular joint dislocation at home and abroad.The specific clinical efficacy of the two procedures and whether there are other risks need to be explored. OBJECTIVE:To compare the short-term postoperative clinical efficacy of arthroscopic TightRope band plate fixation with single tunnel fixation of the coracoid process and tunnel-free suspension fixation of the coracoid process in the treatment of acute type Ⅲ-Ⅴ acromioclavicular joint dislocation. METHODS:A retrospective analysis was performed in 45 patients with acromioclavicular joint dislocation who met the inclusion criteria admitted to the Sixth Affiliated Hospital of Xinjiang Medical University from June 2019 to September 2022,and were divided into coracoid single tunnel fixation group(20 cases)and coracoid tunnel-free suspension fixation group(25 cases)according to the surgical treatment plan.Operation time,incision length,blood loss,Constant-Murley score,visual analogue scale score,the American Shoulder and Elbow Surgeons(ASES)score and intraoperative and postoperative complications of the shoulder joint before operation,3 months after surgery and the last follow-up were compared between the two groups. RESULTS AND CONCLUSION:All patients successfully completed the operation,and there was no important nerve or blood vessel damage during the operation.The operation time of the coracoid tunnel-free suspension fixation group was significantly shorter than that of the coracoid tunnel-free suspension fixation group(P<0.05).There was no significant difference in intraoperative blood loss and incision length between the two groups(P>0.05).All patients were followed up for 12 to 24 months,with an average of(15.29±2.73)months.In the coracoid single tunnel fixation group,at 3 months after operation and the final follow-up,the visual analogue scale score was significantly lower than the preoperative score(P<0.05);Constant-Murley score and ASES score were significantly increased compared with the preoperative values(P<0.05).In the coracoid tunnel-free suspension fixation group,at 3 months after operation and the final follow-up,the visual analogue scale score was significantly lower than the preoperative score(P<0.05);the Constant-Murley score and the ASES score were both significantly higher than the preoperative scores(P<0.05).At 3 months after operation,the Constant-Murley score of the coracoid tunnel-free suspension fixation group was higher than that of the coracoid single tunnel fixation group(P<0.05),while there was no significant difference in visual analogue scale and ASES scores between the two groups(P>0.05).There was also no significant difference in the visual analogue scale,Constant-Murley,and ASES scores between the two groups at the corresponding time points before surgery and at the final follow-up(P>0.05).Intraoperative and postoperative complications:In the coracoid single tunnel fixation group,there was one case of coracoid cortical rupture and fracture during the tunnel drilling during the operation,and one case of a loss of reduction at 3 months after operation,which was repositioned and fixed with hook plate transposition of the coracoacromial ligament.All patients had good acromioclavicular joint function recovery and no re-dislocation at the final follow-up.All patients in the coracoid tunnel-free suspension fixation group did not suffer from coracoid fractures,loss of reduction and other complications during surgery,postoperatively and at the last follow-up.To conclude,these two arthroscopic treatments for acute type Ⅲ-Ⅴ acromioclavicular joint dislocation have the advantages of less trauma,reliable reduction and fixation,and good recovery of shoulder joint function after operation.However,compared with the coracoid single tunnel technique,the coracoid tunnel-free suspension fixation requires shorter time,faster recovery of shoulder joint function in the short term,and avoids the establishment of bone tunnels on the coracoid process,which reduces the probability of iatrogenic fracture of the coracoid process during operation and provides a higher degree of safety.
6.Effects of electroacupuncture on the expression of metabolic enzymes and autophagy genes in gastrocnemius muscle tissues of exercising rats
Rongfa ZHENG ; Weibin MO ; Peng HUANG ; Junji CHEN ; Ting LIANG ; Fangyu ZI ; Guofeng LI
Chinese Journal of Tissue Engineering Research 2025;29(6):1127-1136
BACKGROUND:Acute exercise tends to cause skeletal muscle tissue damage and lipid metabolism disorders in vivo,but the mechanism by which acute exercise combined with electroacupuncture modulates metabolic and autophagic pathways in vivo is unclear. OBJECTIVE:To observe the changes in metabolic enzymes and autophagy levels in skeletal muscle of rats subjected to acute exercise by electroacupuncture at the acupoints of"Zusanli"and"Huantiao." METHODS:Fifty male Sprague-Dawley rats were randomly divided into three groups:quiet control group(n=10),model group(n=20),and reverse electroacupuncture group(n=20).The latter two groups were set up with two time points,i.e.immediate and 3 hours after exercise groups(n=10 per time point).The model group and the reverse electroacupuncture group underwent acute exercise training after adaptive treadmill training.The rats in the reverse electroacupuncture group underwent electroacupuncture treatment(parameters:electroacupuncture on both sides of the rats at the acupoints of"Zusanli"and"Huantiao,"continuous wave,frequency of 2 Hz,intensity of 2 mA,leaving the needle in the body for 30 minutes,once a day for 7 consecutive days)before treadmill training.Bilateral gastrocnemius muscle tissues were taken under anesthesia immediately after exercise and 3 hours after exercise,and hematoxylin-eosin staining was used to observe the histopathological changes of rat skeletal muscle.ELISA kit was used to detect the activities of hepatic lipase,fatty acid synthase,hormone-sensitive lipase,and carnitine palmitoyltransferase 1 in rat skeletal muscle tissues.Immunohistochemistry and western blot were used to detect the changes in the expression of autophagy genes. RESULTS AND CONCLUSION:After hematoxylin-eosin staining,the arrangement of gastrocnemius muscle fibers in the model group was disturbed,swollen and ruptured immediately after exercise and 3 hours after exercise.In the reverse electroacupuncture group,gastrocnemius muscle fibers were tightly arranged and the number of swollen and ruptured cells was greatly reduced immediately after exercise and 3 hours after exercise,and there was no significant difference when compared with the quiet control group.Compared with the quiet control group,the activities of hepatic lipase and fatty acid synthase were lower while the activities of lipoprotein lipase,hormone-sensitive lipase,and carnitine palmitoyltransferase 1 were higher in the model group and the reverse electroacupuncture group 3 hours after exercise(P<0.05 or P<0.01).Compared with the model group,the activities of lipoprotein lipase and carnitine palmitoyltransferase 1 were higher in the reverse electroacupuncture group immediately after exercise(P<0.05),while the activity of lipoprotein lipase was higher and the activity of hormone-sensitive lipase was lower in the reverse electroacupuncture group 3 hours after exercise(P<0.01).Immunohistochemical results showed that compared with the quiet control group,the expression of P62,autophagy-related gene 5 and autophagy-related gene 7 was higher in the model group immediately and 3 hours after exercise,as well as in the reverse electroacupuncture group immediately after exercise(P<0.05 or P<0.01);compared with the model group,the expression of P62 and autophagy-related gene 7 was lower in the reverse electroacupuncture group immediately and 3 hours after exercise(P<0.05).Western blot results showed that the protein expression of P62 and autophagy-related gene 7 in the reverse electroacupuncture group was lower than that in the model group immediately after exercise(P<0.05);the protein expression of Parkin in the model group was higher than that in the quiet control group immediately and 3 hours after exercise(P<0.05);and the protein expression of Parkin in the reverse electroacupuncture group was lower than that in the model group immediately and 3 hours after exercise(P<0.05).To conclude,acute exercise induces disorders,swelling and rupture of gastrocnemius muscle fibers in rats and electroacupuncture on both sides of the acupoints of"Zusanli"and"Huantiao"can improve the level of lipid metabolism and regulate autophagy cells in rat skeletal muscle,preventing the disorders of lipid metabolism and damage of gastrocnemius muscle tissues caused by acute exercise.The mechanism may be closely related to the regulation of autophagy-related factor P62,autophagy-related gene 5,autophagy-related gene 7,and Parkin protein expression to promote the occurrence of autophagy or regulate the autophagy pathway in rat skeletal muscle cells.
7.Different exercise modalities promote functional recovery after peripheral nerve injury
Xiaoxuan ZHAO ; Shuaiyi LIU ; Qi LI ; Zheng XING ; Qingwen LI ; Xiaolei CHU
Chinese Journal of Tissue Engineering Research 2025;29(6):1248-1256
BACKGROUND:Exercise as a form of active rehabilitation can improve the dysfunction caused by peripheral nerve injury,and different exercise modalities target different lesion sites and recovery mechanisms. OBJECTIVE:To comprehensively analyze the application and mechanisms of different exercise modalities in functional recovery from peripheral nerve injury. METHODS:A computerized search was conducted in PubMed and CNKI databases for relevant literature published before January 2024.The search terms used were"peripheral nerve injury,spinal cord,exercise,cerebral cortex,muscle atrophy,mirror therapy,blood flow restriction training"in both English and Chinese.Finally,77 articles were included for review. RESULTS AND CONCLUSION:Peripheral nerve injury can cause systemic pathological changes such as skeletal muscle atrophy,corresponding spinal cord segmental lesions,and sensorimotor cortex remodeling.Aerobic exercise can improve dysfunction by enhancing the immune response,promoting glial cell polarization,and promoting the release of nerve growth factor.Blood flow restriction exercise can regulate the secretion of muscle growth factor,promote muscle growth and enhance muscle strength.Mirror movement has a good effect in activating the cerebral cortex and reducing cortical remodeling.Different exercise modalities have potential benefits in functional recovery after peripheral nerve injury;however,there are still some problems and challenges,such as the choice of exercise modalities,the control of exercise intensity and frequency,and the detailed analysis of mechanisms.
8.Comparison of the clinical efficacy of super pulse thulium laser enucleation of the prostate with "open tunnel" and holmium laser enucleation of the prostate for benign prostatic hyperplasia
Jidong XU ; Ning JIANG ; Jian LI ; Zhikang CAI ; Jianwei LYU ; Chuanyi HU ; Jingcun ZHENG ; Zhonglin CAI ; Huiying CHEN ; Yan GU ; Yuning WANG ; Jiasheng YAN ; Zhong WANG
Journal of Modern Urology 2025;30(1):34-38
[Objective] To compare the clinical efficacy of super pulse thulium laser enucleation of the prostate (SPThuLEP) with "open tunnel" and transurethral holmium laser enucleation of the prostate (HoLEP) in the treatment of benign prostatic hyperplasia (BPH), in order to provide reference for the treatment options of BPH. [Methods] The clinical data of 112 BPH patients treated in our hospital during Jan.2023 and Jul.2023 were retrospectively analyzed, including 65 treated with SPThuLEP with "open tunnel" and 57 with HoLEP.The operation time, postoperative hemoglobin decrease, postoperative bladder irrigation, catheter indwelling time, hospitalization time and complications were compared between the two groups.The changes of maximum urine flow rate (Qmax), international prostate symptom score (IPSS), quality of life score (QoL), postvoid residual (PVR) and prostate-specific antigen (PSA) were compared between the two groups before operation and one month after operation. [Results] All operations were successful without conversion to open or transurethral plasmakinetic resection.The postoperative decrease of hemoglobin in SPThuLEP group was lower than that in HoLEP group [(13.12±6.72) g/L vs. (21.02±6.51) g/L], with statistical difference (P<0.05). There were no significant differences in the operation time [(63.35±15.73) min vs.(61.02±17.55) min], postoperative bladder irrigation time [(1.07±0.45) d vs. (1.06±0.36) d], catheter indwelling time [(2.98±0.56) d vs. (3.01±0.63) d] and hospitalization time [(3.63±0.61) d vs.(3.79±0.76) d] between the two groups (P>0.05). No blood transfusion, secondary bleeding or unplanned hospitalization occurred, and there were no serious complications such as transurethral electroresection syndrome (TURS), urethral stricture and urinary incontinence.One month after operation, the Qmax, IPSS, QoL, PVR and PSA of the two groups were significantly improved compared with those before operation (P<0.05), but with no statistical difference between the two groups (P>0.05). [Conclusion] SPThuLEP with "open tunnel" has comparable efficacy as HoLEP in the treatment of BPH.With advantages of small amount of bleeding and high safety, this minimally invasive technique can be widely popularized in clinical practice.
9.Research progress of super-pulse fiber thulium laser lithotripsy
Tianfu DING ; Zhongyue HUANG ; Zheng XU ; Jianxing LI
Journal of Modern Urology 2025;30(1):82-85
Super-pulse thulium fiber laser (hereinafter referred to as thulium laser) has shown broad application prospects in medical research, especially in the treatment of urinary tract stones.Thulium laser has a thin fiber diameter, which reduces the occupation of the working channel and increases the irrigation water flow.Currently, the peak power reaches up to 500 W, the single pulse energy to 6 J, and the frequency to 2000 Hz.The stone-breaking mechanism of thulium laser is photothermal and photomechanical.Different combinations of energy and frequency will significantly affect the volume of stone fragments. "High energy and low frequency" is more suitable for "fragmentation" stone breaking, while "low energy and high frequency" is more suitable for "powderization" stone breaking.Some in vitro experiments have confirmed that thulium laser has higher stone-breaking efficiency, less stone displacement, and more obvious thermal effects compared to holmium laser.Clinically, thulium laser has been applied to ureteroscopic lithotripsy and percutaneous nephrolithotomy., and the stone free rate and safety are relatively high.This article reviews the stone-breaking principles, in vitro experiments, and clinical applications of thulium laser, aiming to provide better theoretical support and practical experience for the development and application of super-pulse thulium fiber laser in the future.
10.Effect of Yigan Fupi Prescription (抑肝扶脾方) on the AKT/mTOR Pathway in the Colon Tissue of Diarrhea-Type Irritable Bowel Syndrome Model Rats with Liver Stagnation and Spleen Deficiency Pattern
Suting JIU ; Huiying LI ; Yueting SUN ; Songxiang CUI ; Xintian XU ; Hao ZHENG ; Weimin LU
Journal of Traditional Chinese Medicine 2025;66(3):290-299
ObjectiveTo explore the possible mechanism of Yigan Fupi Prescription (抑肝扶脾方, YFP) in treating diarrhea-type irritable bowel syndrome (IBS-D) by investigating the AKT/mTOR signaling pathway. MethodsSixty SD rats were randomly divided into control group, model group, YFP low-, medium-, and high-dose group, and pinaverium bromide group, with 10 rats in each group. All groups but the control group, were subjected to 21 days of tail-clamping stimulation and 14 days of senna leaf gavage to establish a liver stagnation and spleen deficiency-type IBS-D rat model. After successful modeling, the YFP low-, medium-, and high-dose group were administered 0.96, 1.93, and 3.87 g/(kg·d) of the prescription, respectively. The pinaverium bromide group was given 13.5 mg/(kg·d), while the control and model groups were given 10 ml/(kg·d) distilled water. All groups were administered once daily for 14 consecutive days. General conditions of the rats were recorded during the experiment, and after modeling and drug administration, body weight, Bristol stool score, abdominal withdrawal reflex (AWR) score, and histo pathology of colon tissue were observed under HE staining. ELISA was used to detect serum levels of tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). Immunofluorescence was employed to detect the levels of AKT/mTOR pathway-related proteins including phosphorylated AKT (p-AKT)/AKT and phosphorylated mTOR (p-mTOR)/mTOR in the colon tissue. Western Blotting was used to detect the levels of autophagy-related proteins, including UNC-51-like kinase 1 (ULK1), Beclin1 and LC3, and tight junction proteins including Occludin and ZO-1 in the colon tissue. ResultsAfter modeling, compared to the control group, the body weight of rats in the other groups decreased, and Bristol stool scores, as well as AWR scores under 20, 40, 60, and 80 mmHg increased (P<0.05 or P<0.01). After drug administration, compared to the control group, the model group showed reduced body weight, decreased ULK1, Beclin1, LC3Ⅱ/LC3Ⅰ, Occludin, and ZO-1 protein levels in the colon tissue (P<0.05 or P<0.01), and increased Bristol stool scores, AWR scores, serum TNF-α, IL-1β, and IL-6 levels, as well as p-AKT/AKT and p-mTOR/mTOR protein relative expression levels (P<0.05 or P<0.01). Pathological results showed a significant reduction in goblet cells in the upper part of the glandular layer of the colon, with mild inflammatory cell infiltration. The submucosal collagen fibers were dissolved, with unclear boundaries, pale staining, and microvascular congestion and dilation. Compared with the model group, the YFP low-, medium-, and high-dose group and the pinaverium bromide group showed increased body weight, Beclin1, Occludin, and LC3Ⅱ/LC3Ⅰ protein levels (P<0.05 or P<0.01), and decreased Bristol stool scores, AWR scores under 40, 60, and 80 mmHg, serum IL-1β, IL-6, TNF-α levels, and p-AKT/AKT, p-mTOR/mTOR protein relative expression levels (P<0.05 or P<0.01). The pathological morphology of the rats in the YFP groups and pinaverium bromide group showed varying degrees of improvement. Compared with the pinaverium bromide group, the YFP low- and medium-dose group showed increased AWR scores under 20, 40, and 60 mmHg (P<0.05). The YFP low-dose group had reduced TNF-α, IL-1β, and IL-6 levels, and increased p-mTOR/mTOR protein relative expression levels occured in all YFP groups (P<0.05). Compared with the YFP low-dose group, the YFP high-dose group and pinaverium bromide group showed decreased AWR scores under different pressure levels and reduced p-AKT/AKT protein relative expression levels, while the YFP medium- and high-dose group had elevated serum TNF-α, IL-1β levels and reduced p-mTOR/mTOR protein relative expression levels (P<0.05). ConclusionYFP can effectively improve the pathological injury of colon tissue in IBS-D model rats with liver stagnation and spleen deficiency, reduce Bristol stool and AWR scores, and its mechanism may be related to reducing level of inflammatory factors and inhibiting AKT/mTOR pathway-related proteins in colon tissue, thereby enhancing the expression of autophagy-related proteins in the colon tissue.

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