Objective This study reveals the potential mechanism of electroacupuncture in alleviating the inflamma-tory injury of sensory nerves and alleviating the abnormal sensation on the ocular surface in type 2 diabetes rats with dry eye.Methods Healthy male SD rats were induced to establish type 2 diabetes dry eye rat model.Model rats were ran-domly divided into model group,electroacupuncture group,sham-acupuncture group,and fluorometholone group in the 12th week of the experiment.The blank group was established as well,and each group was intervened for 2 weeks.The corneal fluorescein sodium staining(FL),break-up time(BUT),phenol red thread test(PRT),corneal mechanical per-ception threshold(CTT)and blink frequency were measured in each group of rats.After sampling,the expression of neu-ronal injury markers,neuron specific enolase(NSE),neurofilament light chain protein Nf Light(NF-L),phosphorylated microtubule associated protein(Phospho-TAU),as well as inflammatory factors of cysteine protease-1(Caspase-1),inter-leukin-18(IL-18),and interleukin-1β(IL-1β),were observed in the trigeminal ganglion(TG)and spinal cord caudate nu-cleus(SpVc)of each group of rats.Results At the end of the 14th week,compared with the blank group,the FL of the model group,electroacupuncture group,sham-acupuncture group,and fluorometholone group significantly increased,while PRT and BUT significantly decreased.Moreover,the CTT and blink frequency of the model group,sham-acupuncture group,and fluorometholone group significantly decreased(all P＜0.01).Compared with the model group,the electroacu-puncture group and the fluorometholone group showed a decrease in FL,as well as an increase in BUT,PRT,CTT,and blink frequency,with statistically significant differences(all P＜0.05).Compared with the electroacupuncture group,the sham-acupuncture group showed significantly higher FL along with significantly lower values in BUT,PRT,CTT,and blink frequency(all P＜0.05),while the fluorometholone group exhibited decreased FL,PRT,CTT,and blink frequency but in-creased BUT(all P＜0.05).Compared to the sham-acupuncture group,the fluorometholone group exhibited significantly lower FL,but higher BUT and PRT(all P＜0.01).In TG and Sp Vc,compared with the blank group,the expression of NSE,NF-L and Phospho-TAU increased in the model group and the sham-acupuncture group(all P＜0.05);compared with the model group,the expression of NSE,NF-L and Phospho-TAU significantly reduced in the electroacupuncture group(P＜0.01);compared with the electroacupuncture group,the expression of NSE,NF-L and Phospho-TAU increased in the electroacupuncture group and the expression of Phospho-TAU in the sham-acupuncture group also increased(P＜0.05);compared with the sham-acupuncture group,the expression of Phospho-TAU or NF-L significantly reduced in the fluoro-metholone group(all P＜0.05).The Western Blot results indicate that compared with the blank group,the protein expres-sion of Caspase-1,IL-1β,and IL-18 in TG and SpVc of the model group increased;moreover,the expression of Caspase-1 and IL-18 proteins in the sham-acupuncture group also increased(all P＜0.05).Compared with the model group,the ex-pression of Caspase-1,IL-1β,and IL-18 proteins in TG and SpVc reduced in the electroacupuncture group and the fluoro-metholone group,and the expression of IL-1 β proteins in TG and SpVc also reduced in the sham-acupuncture group(all P＜0.05).Compared with the electroacupuncture group,the expression of Caspase-1,IL-1 β,and IL-18 proteins in TG and SpVc increased in the sham-acupuncture group(all P＜0.05).Compared with the sham-acupuncture group,the expression of Caspase-1,IL-1 β,and IL-18 proteins in TG and SpVc reduced in the fluorometholone group(all P＜0.05).Conclusion Electroacupuncture can effectively alleviate the abnormal ocular surface sensation of type 2 diabetes dry eye rats by alleviating the neuroinflammatory injury in TG and SpVc.

Objective This study reveals the potential mechanism of electroacupuncture in alleviating the inflamma-tory injury of sensory nerves and alleviating the abnormal sensation on the ocular surface in type 2 diabetes rats with dry eye.Methods Healthy male SD rats were induced to establish type 2 diabetes dry eye rat model.Model rats were ran-domly divided into model group,electroacupuncture group,sham-acupuncture group,and fluorometholone group in the 12th week of the experiment.The blank group was established as well,and each group was intervened for 2 weeks.The corneal fluorescein sodium staining(FL),break-up time(BUT),phenol red thread test(PRT),corneal mechanical per-ception threshold(CTT)and blink frequency were measured in each group of rats.After sampling,the expression of neu-ronal injury markers,neuron specific enolase(NSE),neurofilament light chain protein Nf Light(NF-L),phosphorylated microtubule associated protein(Phospho-TAU),as well as inflammatory factors of cysteine protease-1(Caspase-1),inter-leukin-18(IL-18),and interleukin-1β(IL-1β),were observed in the trigeminal ganglion(TG)and spinal cord caudate nu-cleus(SpVc)of each group of rats.Results At the end of the 14th week,compared with the blank group,the FL of the model group,electroacupuncture group,sham-acupuncture group,and fluorometholone group significantly increased,while PRT and BUT significantly decreased.Moreover,the CTT and blink frequency of the model group,sham-acupuncture group,and fluorometholone group significantly decreased(all P＜0.01).Compared with the model group,the electroacu-puncture group and the fluorometholone group showed a decrease in FL,as well as an increase in BUT,PRT,CTT,and blink frequency,with statistically significant differences(all P＜0.05).Compared with the electroacupuncture group,the sham-acupuncture group showed significantly higher FL along with significantly lower values in BUT,PRT,CTT,and blink frequency(all P＜0.05),while the fluorometholone group exhibited decreased FL,PRT,CTT,and blink frequency but in-creased BUT(all P＜0.05).Compared to the sham-acupuncture group,the fluorometholone group exhibited significantly lower FL,but higher BUT and PRT(all P＜0.01).In TG and Sp Vc,compared with the blank group,the expression of NSE,NF-L and Phospho-TAU increased in the model group and the sham-acupuncture group(all P＜0.05);compared with the model group,the expression of NSE,NF-L and Phospho-TAU significantly reduced in the electroacupuncture group(P＜0.01);compared with the electroacupuncture group,the expression of NSE,NF-L and Phospho-TAU increased in the electroacupuncture group and the expression of Phospho-TAU in the sham-acupuncture group also increased(P＜0.05);compared with the sham-acupuncture group,the expression of Phospho-TAU or NF-L significantly reduced in the fluoro-metholone group(all P＜0.05).The Western Blot results indicate that compared with the blank group,the protein expres-sion of Caspase-1,IL-1β,and IL-18 in TG and SpVc of the model group increased;moreover,the expression of Caspase-1 and IL-18 proteins in the sham-acupuncture group also increased(all P＜0.05).Compared with the model group,the ex-pression of Caspase-1,IL-1β,and IL-18 proteins in TG and SpVc reduced in the electroacupuncture group and the fluoro-metholone group,and the expression of IL-1 β proteins in TG and SpVc also reduced in the sham-acupuncture group(all P＜0.05).Compared with the electroacupuncture group,the expression of Caspase-1,IL-1 β,and IL-18 proteins in TG and SpVc increased in the sham-acupuncture group(all P＜0.05).Compared with the sham-acupuncture group,the expression of Caspase-1,IL-1 β,and IL-18 proteins in TG and SpVc reduced in the fluorometholone group(all P＜0.05).Conclusion Electroacupuncture can effectively alleviate the abnormal ocular surface sensation of type 2 diabetes dry eye rats by alleviating the neuroinflammatory injury in TG and SpVc.

Objective To investigate the mechanism by which Erdong Xiaoke Formula(EDXKF)regulates lacrimal gland autophagy in type 2 diabetes-induced dry eye rats through the peroxisome proliferator-activated receptor γ(PPARγ)/mammalian target of rapamycin(mTOR)signal pathway.Methods Healthy male SD rats were fed with high-sugar and high-fat chow and then injected intraperitoneally 30 mg·kg-1 streptozotocin to construct type 2 diabetes-induced dry eye rat models.Healthy male SD rats were selected as a blank group.Type 2 diabetes rats were randomly divided into a model group,a Traditional Chinese Medicine(TCM)group(given 11 g·kg-1 of EDXKF through gavage),an antagonist group(given 2 mg·kg-1 PPARγ antagonist through intraperitoneal injection),and a TCM plus antagonist group(given 11 g·kg-1 EDXKF through gavage and 2 mg·kg-1 PPARγ antagonist through intraperitoneal injection).Fasting blood glucose(FBG),corneal fluorescein(FL)staining,tear film break-up time(BUT)and phenol red cotton thread test(Prtt)were examined before modeling,after modeling,and after intervention.The weight of the lacrimal gland was compared among different groups after sampling.Hematoxylin and eosin(HE)staining was performed to analyze lacrimal gland hismorphol-ogy.The expression of microtubule-associated protein 1 light chain 3(LC3)and Sequestosome-1(p62)in the lacrimal gland was examined by immunofluorescence.Western blot was used to detect the expression of PPARγ,p62,LC3 Ⅱ,LC3 I,mTOR and p-mTOR in the lacrimal gland.Results FBG levels in the antagonist group were significantly increased after intervention,compared with those in the TCM group(P＜0.01).Ocular surface examination showed that compared with the model group,the TCM group had increased BUT and Prtt scores and decreased FL scores,the antagonist group had de-creased BUT and Prtt scores and increased FL scores(all P＜0.05).Compared with the antagonist group,the TCM plus antagonist group showed increased BUT and Prtt scores and decreased FL scores(all P＜0.05).The weight analysis of lac-rimal glands revealed that the lacrimal gland weight increased in the TCM group and decreased in the antagonist group,compared with that in the model group(all P＜0.01).The lacrimal gland weight in the TCM plus antagonist group was higher than that in the antagonist group(P＜0.01).HE staining of the lacrimal gland showed atrophy of glandular lobules,increased fusion and expansion of follicles,and dense distribution of nuclei in model and antagonist groups.These symp-toms were more obvious in the antagonist group.Compared with antagonist and TCM plus antagonist groups,the TCM group showed improved symptoms,with tightly arranged follicles,partial atrophy and fusion,and a small amount of expan-sion.Immunofluorescence staining for detecting the average fluorescence intensity of LC3 and p62 showed that compared with the model group,the TCM group had increased LC3 levels and decreased p62 levels,and the antagonist group had de-creased LC3 levels and increased p62 levels(all P＜0.01).Compared with those in the antagonist group,LC3 levels were increased and p62 levels were decreased in the TCM plus antagonist group(both P＜0.01).Western blot results showed that compared with the model group,the TCM group had increased PPARγ and LC3 levels and decreased p62 and p-mTOR/mTOR levels.PPARγ and LC3 levels were decreased and p62 and p-mTOR/mTOR levels were increased in the antagonist group,compared with those in the model group.The TCM plus antagonist group had higher PPARγ and LC3 levels and low-er p62 and p-mTOR/mTOR levels than the antagonist group(all P＜0.05).Conclusion EDXKF can regulate the PPARy/mTOR signal pathway to promote lacrimal gland autophagy and thus alleviate dry eyes in type 2 diabetes rats.

Objective:To explore the structural equation model to explore the levels of work-related musculoskeletal disorders (WMSDs) and various risk factors in the feet and ankle of China's occupational population, providing scientific basis for for preventing WMSDs in feet and ankles.Methods:Data of 73497 national occupational epidemiological cases were selected from June 2018 to December 2023 used the Chinese version of the Electronic Questionnaire on Musculoskeletal Disorders. The adverse ergonomic factors and their source classification standard and confirmatory factor analysis were used to investigate foot and ankle WMSDs and their related risk factors (including individual factors, work organization, work posture, work type, fatigue, etc.) in key occupational groups in China, and structural equation model hypothesis, fitting, verification, and path and intermediary effect analysis were carried out. The model fit evaluation indexes included Chi-square specific degrees of freedom ( χ2/ df), gauge fit index (NFI), Tucker Lewis index (TLI), goodness of Fit index (GFI), adjusted Goodness of Fit index (AGFI) and approximate root mean square error (RMSEA) . Results:A total of 73497 occupational workers were surveyed, with local muscle fatigue and WMSDs incidence rates in the feet and ankles being 17.17% and 12.06%, respectively. The fitting index of the adjusted structural equation model basically meets the standard (GFI=1, AGFI=1, RMESA=0.042, NFI=0.716, TLI=0.663). The top three factors affecting feet and ankle WMSDs are feet and ankle muscle fatigue, work type, and work organization, with standardized path coefficients of 0.221, 0.105, and 0.095, respectively. The top two factors affecting feet and ankle muscle fatigue are work organization and work type, with standardized path coefficients of 0.548 and 0.383, respectively. Feet and ankle muscle fatigue, work type, work organization, and work posture have a direct effect on feet and ankle WMSDs, with effect values of 0.221, 0.105, 0.095, and 0.077, respectively. The organization and type of work can also have indirect effects through feet and ankle muscle fatigue, with effect values of 0.121 and 0.084, respectively.Conclusion:Feet and ankle muscle fatigue has a direct impact on WMSDs, and plays a mediating role between ankle and ankle WMSDs caused by work organization and work type. Feet and ankle muscle fatigue is an important pathway leading to feet and ankle WMSDs. It is recommended that employers and managers detect job fatigue early and take corresponding prevention and intervention measures, which can play a key role in preventing feet and ankle WMSDs.

Objective:To explore the structural relationship between WMSDs in the upper limbs and various risk factors in the occupational population in China, based on a large sample epidemiological survey and structural equation analysis, and to establish a structural equation model, so as to lay a foundation for the prevention and control of such diseases.Methods:The Chinese version of the Musculoskeletal Disorders Electronic Questionnaire was used to conduct a nationwide survey on the prevalence of WMSDs in the upper extremity. Six factors related to WMSDs in the upper extremity were extracted by the classification standard of adverse ergonomic factors and their source and confirmatory factor analysis, including work organization, work type, upper extremity work posture, individual factors, upper extremity fatigue and upper extremity WMSDs. The structural equation analysis was carried out and the structural equation model was established.Results:The incidence of WMSDs and fatigue in the upper limbs was 24.44% and 43.76%, respectively. The adjusted structural equation model fitting indicators were generally up to the standard (GFI=1.000, AGFI=1.000, RMSEA=0.043, NFI=0.808, TLI=0.784) . The four exogenous latent variables of work organization, work type, upper limb work posture and individual factors were correlated. There was a strong positive correlation between job type and upper limb work posture ( r=0.865) , a moderate positive correlation between work organization and job type and upper limb work posture ( r=0.570, 0.490) , and a weak negative correlation between individual factors and the other three exogenous latent variables. Upper limb work posture and individual factors had direct effects on upper limb WMSDs, and the effect coefficients were 0.10 and 0.06, respectively. Upper limb fatigue played a mediating role between work organization, work type, upper limb work posture and upper limb WMSDs. The effect coefficient was 0.46, and the composition ratios of indirect effects were 100.0%, 100.0%, and 38.3%, respectively. The direct path effect of upper limb work posture, individual factors and upper limb WMSDs was weaker than the mediating path through upper limb fatigue. Conclusion:When carrying out the prevention and control of upper limbWMSDs, it is necessary to comprehensively consider the pathogenesis path of upper limb muscle fatigue and upper limb WMSDs caused by work organization, work type, and upper limb work posture, so as to provide theoretical reference for improving the prevention and control level of such diseases.

Objective:To explore the structural equation model to explore the levels of work-related musculoskeletal disorders (WMSDs) and various risk factors in the feet and ankle of China's occupational population, providing scientific basis for for preventing WMSDs in feet and ankles.Methods:Data of 73497 national occupational epidemiological cases were selected from June 2018 to December 2023 used the Chinese version of the Electronic Questionnaire on Musculoskeletal Disorders. The adverse ergonomic factors and their source classification standard and confirmatory factor analysis were used to investigate foot and ankle WMSDs and their related risk factors (including individual factors, work organization, work posture, work type, fatigue, etc.) in key occupational groups in China, and structural equation model hypothesis, fitting, verification, and path and intermediary effect analysis were carried out. The model fit evaluation indexes included Chi-square specific degrees of freedom ( χ2/ df), gauge fit index (NFI), Tucker Lewis index (TLI), goodness of Fit index (GFI), adjusted Goodness of Fit index (AGFI) and approximate root mean square error (RMSEA) . Results:A total of 73497 occupational workers were surveyed, with local muscle fatigue and WMSDs incidence rates in the feet and ankles being 17.17% and 12.06%, respectively. The fitting index of the adjusted structural equation model basically meets the standard (GFI=1, AGFI=1, RMESA=0.042, NFI=0.716, TLI=0.663). The top three factors affecting feet and ankle WMSDs are feet and ankle muscle fatigue, work type, and work organization, with standardized path coefficients of 0.221, 0.105, and 0.095, respectively. The top two factors affecting feet and ankle muscle fatigue are work organization and work type, with standardized path coefficients of 0.548 and 0.383, respectively. Feet and ankle muscle fatigue, work type, work organization, and work posture have a direct effect on feet and ankle WMSDs, with effect values of 0.221, 0.105, 0.095, and 0.077, respectively. The organization and type of work can also have indirect effects through feet and ankle muscle fatigue, with effect values of 0.121 and 0.084, respectively.Conclusion:Feet and ankle muscle fatigue has a direct impact on WMSDs, and plays a mediating role between ankle and ankle WMSDs caused by work organization and work type. Feet and ankle muscle fatigue is an important pathway leading to feet and ankle WMSDs. It is recommended that employers and managers detect job fatigue early and take corresponding prevention and intervention measures, which can play a key role in preventing feet and ankle WMSDs.

Objective:To explore the structural relationship between WMSDs in the upper limbs and various risk factors in the occupational population in China, based on a large sample epidemiological survey and structural equation analysis, and to establish a structural equation model, so as to lay a foundation for the prevention and control of such diseases.Methods:The Chinese version of the Musculoskeletal Disorders Electronic Questionnaire was used to conduct a nationwide survey on the prevalence of WMSDs in the upper extremity. Six factors related to WMSDs in the upper extremity were extracted by the classification standard of adverse ergonomic factors and their source and confirmatory factor analysis, including work organization, work type, upper extremity work posture, individual factors, upper extremity fatigue and upper extremity WMSDs. The structural equation analysis was carried out and the structural equation model was established.Results:The incidence of WMSDs and fatigue in the upper limbs was 24.44% and 43.76%, respectively. The adjusted structural equation model fitting indicators were generally up to the standard (GFI=1.000, AGFI=1.000, RMSEA=0.043, NFI=0.808, TLI=0.784) . The four exogenous latent variables of work organization, work type, upper limb work posture and individual factors were correlated. There was a strong positive correlation between job type and upper limb work posture ( r=0.865) , a moderate positive correlation between work organization and job type and upper limb work posture ( r=0.570, 0.490) , and a weak negative correlation between individual factors and the other three exogenous latent variables. Upper limb work posture and individual factors had direct effects on upper limb WMSDs, and the effect coefficients were 0.10 and 0.06, respectively. Upper limb fatigue played a mediating role between work organization, work type, upper limb work posture and upper limb WMSDs. The effect coefficient was 0.46, and the composition ratios of indirect effects were 100.0%, 100.0%, and 38.3%, respectively. The direct path effect of upper limb work posture, individual factors and upper limb WMSDs was weaker than the mediating path through upper limb fatigue. Conclusion:When carrying out the prevention and control of upper limbWMSDs, it is necessary to comprehensively consider the pathogenesis path of upper limb muscle fatigue and upper limb WMSDs caused by work organization, work type, and upper limb work posture, so as to provide theoretical reference for improving the prevention and control level of such diseases.

Objective To investigate the mechanism by which Erdong Xiaoke Formula(EDXKF)regulates lacrimal gland autophagy in type 2 diabetes-induced dry eye rats through the peroxisome proliferator-activated receptor γ(PPARγ)/mammalian target of rapamycin(mTOR)signal pathway.Methods Healthy male SD rats were fed with high-sugar and high-fat chow and then injected intraperitoneally 30 mg·kg-1 streptozotocin to construct type 2 diabetes-induced dry eye rat models.Healthy male SD rats were selected as a blank group.Type 2 diabetes rats were randomly divided into a model group,a Traditional Chinese Medicine(TCM)group(given 11 g·kg-1 of EDXKF through gavage),an antagonist group(given 2 mg·kg-1 PPARγ antagonist through intraperitoneal injection),and a TCM plus antagonist group(given 11 g·kg-1 EDXKF through gavage and 2 mg·kg-1 PPARγ antagonist through intraperitoneal injection).Fasting blood glucose(FBG),corneal fluorescein(FL)staining,tear film break-up time(BUT)and phenol red cotton thread test(Prtt)were examined before modeling,after modeling,and after intervention.The weight of the lacrimal gland was compared among different groups after sampling.Hematoxylin and eosin(HE)staining was performed to analyze lacrimal gland hismorphol-ogy.The expression of microtubule-associated protein 1 light chain 3(LC3)and Sequestosome-1(p62)in the lacrimal gland was examined by immunofluorescence.Western blot was used to detect the expression of PPARγ,p62,LC3 Ⅱ,LC3 I,mTOR and p-mTOR in the lacrimal gland.Results FBG levels in the antagonist group were significantly increased after intervention,compared with those in the TCM group(P＜0.01).Ocular surface examination showed that compared with the model group,the TCM group had increased BUT and Prtt scores and decreased FL scores,the antagonist group had de-creased BUT and Prtt scores and increased FL scores(all P＜0.05).Compared with the antagonist group,the TCM plus antagonist group showed increased BUT and Prtt scores and decreased FL scores(all P＜0.05).The weight analysis of lac-rimal glands revealed that the lacrimal gland weight increased in the TCM group and decreased in the antagonist group,compared with that in the model group(all P＜0.01).The lacrimal gland weight in the TCM plus antagonist group was higher than that in the antagonist group(P＜0.01).HE staining of the lacrimal gland showed atrophy of glandular lobules,increased fusion and expansion of follicles,and dense distribution of nuclei in model and antagonist groups.These symp-toms were more obvious in the antagonist group.Compared with antagonist and TCM plus antagonist groups,the TCM group showed improved symptoms,with tightly arranged follicles,partial atrophy and fusion,and a small amount of expan-sion.Immunofluorescence staining for detecting the average fluorescence intensity of LC3 and p62 showed that compared with the model group,the TCM group had increased LC3 levels and decreased p62 levels,and the antagonist group had de-creased LC3 levels and increased p62 levels(all P＜0.01).Compared with those in the antagonist group,LC3 levels were increased and p62 levels were decreased in the TCM plus antagonist group(both P＜0.01).Western blot results showed that compared with the model group,the TCM group had increased PPARγ and LC3 levels and decreased p62 and p-mTOR/mTOR levels.PPARγ and LC3 levels were decreased and p62 and p-mTOR/mTOR levels were increased in the antagonist group,compared with those in the model group.The TCM plus antagonist group had higher PPARγ and LC3 levels and low-er p62 and p-mTOR/mTOR levels than the antagonist group(all P＜0.05).Conclusion EDXKF can regulate the PPARy/mTOR signal pathway to promote lacrimal gland autophagy and thus alleviate dry eyes in type 2 diabetes rats.

Objective To observe the effect of electroacupuncture on corneal Toll-like receptor 4(TLR4)-mediated inflammatory signaling pathway in diabetic dry eye rats and to explore the mechanism of electroacupuncture in the treat-ment of diabetic dry eye.Methods A type 2 diabetic rat model was established in 32 healthy male Sprague-Dawley(SD)rats by intraperitoneal injection of streptozotocin(30 mg·kg-1)for 12 weeks after feeding with high-sugar and high-fat di-et for 4 weeks.Twenty-five successfully modeled diabetic dry eye rats were randomly divided into the model group(non-in-tervention),electroacupuncture group(the"Jingming","Cuanzhu","Sizhukong","Taiyang"and"Tongziliao"acupoints were treated with acupuncture,and then"Cuanzhu"and"Tongzilao"acupoints were treated with electroacupuncture,15 min for each time,once a day),sham acupuncture group(blunt-tip needle pricking was performed at the same acupoints as the electroacupuncture group),and fluorometholone group(1 g·L-1fluorometholone eye drops were used in both eyes at 8 o'clock,13 o'clock,and 18 o'clock,1 drop each time),with 6 rats in each group,lasting for 2 weeks.Another 6 healthy male SD rats were selected as a blank group.Random blood glucose,tear film breakup time(BUT),tear secre-tion,corneal fluorescein staining(FL)score,and corneal touch threshold(CTT)of rats in each group were detected be-fore modeling,after modeling,and after the intervention.Hematoxylin and Eosin(HE)staining was performed to observe the corneal morphologic changes in each group.Immunofluorescence histochemical staining was adopted to detect the cor-neal TLR4-positive expression in each group.The TLR4,phosphorylated nuclear factor-kappa P65(P-NF-KB P65),inter-leukin(IL)-1β,and IL-18 expression levels in the cornea were detected by Western blot.Results After modeling,com-pared with the blank group,BUT,tear secretion and CTT decreased and FL increased in all experimental groups,and the differences were statistically significant(all P＜0.01).After the intervention,compared with the model group,FL de-creased,and BUT,tear secretion and CTT increased in the electroacupuncture group,and the differences were statistically significant(all P＜0.05).Corneal HE staining showed that after the intervention,the corneal surface of rats in the model group and sham acupuncture group was not smooth,and the corneal epithelial cells were thickened and disorganized;the corneal surface of rats in the electroacupuncture group and fluorometholone group was smooth,and the corneal epithelial cells were arranged neatly.After the intervention,compared with the blank group,the corneal TLR4 expression of rats in all other groups was elevated,and the differences were statistically significant(all P＜0.05);compared with the model group,the corneal TLR4 expression of rats in the electroacupuncture group and fluorometholone group was reduced(both P＜0.01).Compared with the blank group,corneal TLR4,P-NF-κBP65,IL-1β and IL-18 protein expressions increased in the model group and sham acupuncture group(all P＜0.05);compared with the model group,these expressions decreased in the electroacupuncture group and the fluorometholone group(all P＜0.05).Conclusion Electroacupuncture can im-prove the ocular surface signs and inhibit the expressions of TLR4,P-NF-κB P65,IL-1 β,and IL-18 in the cornea of type 2 diabetic dry eye rats,and its mechanism of action may be related to the regulation of TLR4-mediated inflammatory signaling pathway,which inhibits ocular surface inflammation in diabetic dry eye rats.

Objective To reveal the potential mechanism of electroacupuncture regulating the neuroinflammatory signaling pathway mediated by triggering receptor expressed on myeloid cells 2(TREM2)in trigeminal ganglion(TG)and caudate nucleus of spinal trigeminal nucleus(SpVc)of rats with type 2 diabetes and dry eyes and alleviating ocular surface sensory abnormalities.Methods Healthy male Sprague-Dawley(SD)rats were induced by intraperitoneal injection of 10 g·L-1 streptozotocin to establish a type 2 diabetes dry eye rat model after 4 weeks of high-glucose and high-fat diets.In the 12th week of the experiment,successfully modeled rats were randomly divided into the model group,electroacupunc-ture group,sham-acupuncture group and fluorometholone group,and healthy male SD rats fed with normal diets were se-lected as the blank group.Rats in all groups were intervened for 2 weeks.The corneal fluorescein sodium staining(FL),tear secretion[detected by phenol red thread(PRT)test],tear film break-up time(BUT),and corneal touch threshold(CTT)in each group were measured before,after modeling,and after the intervention.The changes in tissue morphology of TG and SpVc and TREM2 positive expression sites in each group were observed.The messenger ribonucleic acid(mR-NA)expression of TREM2,interleukin-18(IL-18)and interleukin-1β(IL-1β)of TG and Sp Vc in each group were detec-ted.Results After modeling,compared with the blank group,the FL scores of rats significantly increased and PRT,BUT and CTT significantly decreased in other groups(all P＜0.01).After the intervention,compared with the model group,the electroacupuncture group and fluorometholone group showed significant reductions in FL scores and significant increases in PRT,BUT and CTT(all P＜0.01);there was no statistically significant difference in these indicators in the sham-acupuncture group(all P＞0.05).Compared with the electroacupuncture group,the PRT and CTT of rats in the flu-orometholone group and sham-acupuncture group were significantly reduced,the FL score significantly increased and BUT significantly decreased in the sham-acupuncture group(all P＜0.01),and there was no statistically significant difference in FL score and BUT of rats in the fluorometholone group(both P＞0.05).Compared with the sham-acupuncture group,the fluorometholone group showed a decrease in FL score and an increase in PRT,BUT and CTT in rats,with statistically dif-ferent significances(all P＜0.05).The immunofluorescence double-labeling assay showed a positive expression of TREM2 in activated microglia of TG and SpVc in the model group.The reverse transcription-polymerase chain reaction of TG and SpVc showed that compared with the blank group,the TREM2 mRNA expression in the TG and SpVc of the model group,electroacupuncture group,fluorometholone group and sham-acupuncture group decreased,the IL-18 and IL-1β mRNA in TG and SpVc of the model group,IL-18 and IL-1β mRNA in TG and IL-1β mRNA in SpVc of the sham-acupuncture group increased(all P＜0.05);compared with the model group,the TREM2 mRNA expression increased and IL-18 and IL-1β mR-NA expression decreased in the TG and SpVc of the electroacupuncture group and fluorometholone group(all P＜0.05).Compared with the electroacupuncture group,the TREM2 mRNA expression decreased and IL-18,IL-1β mRNA expression increased in TG and SpVc of the sham-acupuncture group(all P＜0.05),and no significant difference was found in TREM2,IL-18 and IL-1β mRNA expression in TG and SpVc of the fluorometholone group(all P＞0.05).Compared with the sham-acupuncture group,the TREM2 mRNA expression in TG and SpVc increased in the fluorometholone group,while the IL-18 mRNA expression in SpVc decreased(all P＜0.05);the IL-1β mRNA in TG and SpVc and the IL-18 mRNA in TG of the fluorometholone group showed no statistically significant difference(all P＞0.05).Conclusion Electroacupunc-ture can effectively alleviate the ocular surface sensory abnormalities of rats with type 2 diabetes and dry eyes by regulating the inflammatory signaling pathway mediated by TREM2 in TG and SpVc.