Discussion On The Therapeutic Effect And Mechanism Of Acupuncture In The Treatment Of Age Related Hearing Loss With Cognitive Dysfunction Under The Guidance Of The Method Of Strengthening The Spleen And Nourishing The Kidney Based On Data Mining

1 Objective Based on data mining to summarize the rules of acupuncture treatment of hearing loss in the past two decades,to study the clinical efficacy of the method of strengthening the spleen and nourishing the kidney in the treatment of age related hearing loss(ARHL)with mild cognitive impairment(MCI),and setting up animal experiments under the guidance of the treatment to explore the possible mechanism of ARHL with cognitive dysfunction using acupuncture method,to provide clinical support for ARHL with MCI,and provides a reference for basic research.2 Methods2.1 Modern literature data mining of acupuncture for hearing loss By screening all the clinical electronic literature on acupuncture and moxibustion in the treatment of hearing loss in the past 20 years in the 4 databases of CNKI,VIP Chinese scientific journals,Wanfang academic journals and Chaoxing Duxiu academic search platform,set up the search term of "deafness" OR "hearing loss" AND " Acupuncture " OR "Moxibustion" OR "Electro-Acupuncture" OR "Acupoint Embedding" OR "Acupoint Injection" to carry out a literature search for the synonyms of the subject words,the language of the journal is limited to Chinese,and the retrieval time starts in 1st January,2000 to December 31 st,2021.According to the set inclusion and exclusion criteria,the sorted literature will divided into four sub-directions: acupuncture,acupoint catgut embedding,acupoint injection and moxibustion.We will take Frequencies Analysis,Association Analysis,Cluster Analysis and Factor Analysis to summarize the rules.2.2 Clinical efficacy study of acupuncture in the treatment of ARHL with MCI guided by the method of strengthening the spleen and nourishing the kidneyIn this study,ARHL patients with MCI has been selected as the research objects,according to the randomized controlled research method 72 patients were randomly divided into the test group(TG)and the control group(CG)according to 1:1.It has been obtained The ethics number(2021SL-027)and registered in the Chinese Clinical Trials Registry(Chi CTR2100050341).The TG was given acupuncture combined with basic treatment,and the CG was given basic treatment.Take the Audiology and Recognition Test in 0 months,1 month,2 months,3 months after treatment and 1month follow up in a total of 5 time windows.Monitoring indicators include pure tone hearing threshold(PTA),word recognition score(WRS),Montreal assessment scale(Mo CA)and adverse events.SPSS 25.0 were used for statistical analysis of the collected data,the differences in curative effect within and between groups at different time points has been compared,and the curative effect and safety of acupuncture guided by the method of strengthening the spleen and nourishing the kidney in the treatment of ARHL with MCI were objectively evaluated.2.3 Mechanism of D-galactose induced ARHL and cognitive decline in mice treated with acupuncture under the guidance of spleen-strengthening and kidney-nourishing method36 aged 7 month C57BL/6J mice were randomly divided into three groups(acupuncture group(AG),model group(MG)and blank group(BG)).D-gal were injected into the nape of the neck in The AG and MG last 4 weeks,after modeling 2weeks,the AG was given acupuncture intervention for 4 consecutive weeks,and the MG and the BG were given bundled treatment for 4 consecutive weeks,once a day,30 min a time,after intervention the three groups of mice underwent ABR test,Morris maze test and the auditory cortex and hippocampus got HE staining and TUNEL test.Statistical analysis was performed using SPSS 25.0 to compare the differences in indicators between groups.3 Results3.1 Modern literature data mining of acupuncture for hearing loss(1)A total of 314 effective prescriptions were collected from 270 articles,including234 for acupuncture,3 for catgut embedding,57 for acupoint injection,and 20 for moxibustion.108 acupoints were used,with a cumulative citation frequency of 2,540.There are 8 acupoints with a citation frequency of ≥100,namely Tinggong(SI19)(249),Yifeng(TE17)(235),Tinghui(GB2)(226),Ermen(TB21)(191),Zhongzhu(TE3)(125),Taixi(KI3)(110),Waiguan(TE5)(109),Taichong(LR1)(106).The cumulative frequency of high-frequency acupoints(frequency ≥30)reached 81.10%.Analyze the acupoint feature,location and meridian tropism of high-frequency acupoints,the top 3 meridian are The hand Shaoyang meridian(25.98%),the foot Shaoyang meridian(18.50%)and the hand Taiyang meridian(9.80%).The head and neck(42.91%)and lower limbs(23.39%)were the main location,the top 3 acupoint feature are the five shu acupoints(28.93%),confluential Points(28.84%)and non-specific acupoints(25.36%).(2)Association analysis: High-frequency acupoints(frequency ≥ 30)were subjected to association analysis,and obtained 257 association rules.The minimum support rate was 10%,the maximum support rate was 74.29%,the minimum confidence was 80%,and the maximum confidence was 100%.Yifeng(TE17)--Tinggong(SI19)and Tinggong(SI19)--Tinghui(GB2)are the top two support rate.The correlation analysis of high-frequency acupoints returning to meridians obtained 120 association rules,the minimum support rate degree was10.61%,the maximum support rate degree was 91.96%,the minimum confidence degree rate was 80.28%,and the maximum confidence degree was 100%.The hand Shaoyang Sanjiao Meridian--the hand Taiyang small intestine Meridian and the hand Shaoyang Sanjiao Meridian--the foot Shaoyang gallbladder Meridian have the highest support rate.(3)Cluster analysis: Cluster analysis was performed on high-frequency acupoints(frequency ≥30),and they were clustered into eight categories:(a): Tinggong(SI19),Yifeng(TE17);(b): Tinghui(GB2),Ermen(TB21);(c): Zhongzhu(TE3),Xiaxi(GB43);(d): Taixi(KI3),Shensu(BL23),Fenglong(ST40),Qiuxu(GB40);(e):Zusanli(ST36),Sanyinjiao(SP6),Taichong(LR1),Waiguan(TE5),Hegu(LI4),Xuehai(SP10);(f): Fengchi(GB20);(g): Baihui(GV20),Wangu(GB12);(h):Zulinqi(GB41).Combined with frequency analysis,category(a): Tinggong(SI19),Yifeng(TE17)and category(b): Tinghui(GB2)and Ermen(TB21)are the main acupoints,and the rest are auxiliary points.(4)Factor analysis: Perform factor analysis on high-frequency acupoints(frequency≥30),set the absolute value of the minimum load table to 0.40,and obtain 8 common factors,namely F1: Hegu(LI4),Zusanli(ST36),Taichong(LR1),Waiguan(TE5),Sanyinjiao(SP6),Xuehai(SP10);F2: Qiuxu(GB40),Fonglong(ST40),Shensu(BL23);F3: Tinghui(GB2),Ermen(TB21);F4:Yifeng(TE17),Tinggong(SI19),Taixi(KI3);F5: Zhongzhu(TE3),Xiaxi(GB43);F6: Zu Linqi(GB41);F7: Wangu(GB12),Baihui(GV20);F8: Fengchi(GB20).3.2 Clinical efficacy study of acupuncture in the treatment of ARHL with MCI guided by the method of strengthening the spleen and nourishing the kidney(1)The method of strengthening the spleen and nourishing the kidney in the treatment of ARHL with MCI is effective in both hearing and cognitive function aspects.(2)Hearing aspect:PTA:With the prolongation of the treatment period,the threshold value of the test group(TG)at 1000 Hz and 2000 Hz frequency decreased,and there was a statistically significant difference compared with the control group(CG)in the second month after treatment(P<0.05).The effective rate of the two groups gradually increased,the highest value was reached in 3 months,the highest rate in the TG was26.67%,and the highest rate in the CG was 7.4%,but there was no statistical difference in the efficacy between the two groups(P>0.05).Mild to moderate hearing loss,flat hearing curve,and no underlying disease have a better prognosis.WRS: The speech recognition rate scores in both groups were improved after treatment,the improvement time of the TG was earlier than that of the CG,and the speed was higher than that of the CG,the difference in speech recognition rate scores was statistically significant(P<0.05).There was a significant difference in the efficacy between the two groups after 2 month treatment(P<0.05).The efficacy of the TG was significantly better than that of the CG,and the gap gradually increased.The highest effective rate of the TG was 40%.With the prolongation of the treatment period,the degree of speech recognition in the TG gradually decreased,in the two groups at 3 months after treatment and 1 month of follow-up,and the degree in the experimental group was significantly lighter than that in the control group(P<0.05).(3)Cognitive aspects: The total Mo CA score of the TG increased with the prolongation of the treatment period,and was statistically significant compared with the CG after 2 month treatment.The TG started to have statistical differences in memory--delayed memory and attention after 2 month treatment(P<0.05),and after 3month treatment,there were statistical differences in the two aspects of visual space executive ability and language(P<0.05),indicating that the patient’s memory--delayed memory,attention,language and visual space executive ability can be improved after treatment,among which memory--delayed memory,attention changes the earliest.3.3 Mechanism of D-gal induced ARHL and cognitive decline in mice treated with acupuncture under the guidance of spleen-strengthening and kidney-nourishing method(1)Subcutaneous injection of 1000 mg/Kg·day D-gal into the nape of the neck of7-month C57BL/6J mice for 4 consecutive weeks can successfully create a mouse model of ARHL with cognitive impairment.There was no significant difference in the ABR threshold of this model.in the behavioral Morris water maze,the latency was prolonged,the number of crossing platforms and the number of crossing platforms were reduced,compared with the blank group,P<0.05,which was statistically significant.Auditory cortex and hippocampus HE Staining showed that the pathological damage was serious,with neuronal patchy or diffuse degeneration and necrosis,loose interstitial structure,telangiectasia,gliosis and other pathological damages,and the number of pyramidal cell layers in the CA3 area of the hippocampus was reduced,see the CA1 area Pyramidal cells pyknosis,hyper-chromatic nuclei.A large number of apoptotic neurons were seen in the auditory cortex of the model group,compared with the blank group,P=0.000,which was statistically significant.There were a large number of dark brown-yellow positive neurons in the hippocampus CA1 and CA3 of the model group.Compared with the blank group,P =0.005,which is statistically significant.(2)In terms of ABR detection: there was no statistical significance between the model group and the blank group in each frequency,click,and average threshold value,indicating that the changes in peripheral hearing were not obvious after subcutaneous injection of D-gal into the nape of the neck,and the central hearing changes were the main changes.Compared before and after the blank group,although each frequency P>0.05,the threshold values of 12 KHz,18KHz,average and click levels were increased,indicating that the hearing of mice was further reduced.Compared with the model group before and after modeling,the threshold values of each frequency increased faster in the model group than in the blank group,suggesting that the subcutaneous injection of D-gal into the neck and back may accelerate the aging of peripheral hearing.After intervention,the threshold values of 12 KHz,18KHz and click levels in the acupuncture group decreased compared with those before modeling,suggesting that acupuncture may improve the hearing of mice at these frequencies,and at the 12 KHz level,compared with the model group,P=0.031 < 0.05,which is comparable,indicating that acupuncture can improve the progress of hearing decline after subcutaneous injection of D-gal.(3)The method of strengthening the spleen and nourishing the kidney can improve the central hearing impairment and cognitive impairment in ARHL with MCI mice.In terms of ABR,the acupuncture group and the model group had statistical significance at the 12 KHz level,P<0.05.Compared with the model group,the damage of the auditory cortex and hippocampus of HE staining was significantly reduced,and the number of cortical neuron lesions was reduced.The TUNEL test for auditory cortex and hippocampus in acupuncture group shows that the number of apoptotic neurons in the auditory cortex was higher than that in the blank group,but less than that in the model group.Compared with the model group,P=0.01<0.05,which was statistically significant.acupuncture can reduce the Cortical neuron apoptosis to reduce the damage of D-gal to the auditory center and improve the progress of hearing in presbycusis.There were a large number of dark brown-yellow positive neurons in the hippocampus CA1 and CA3 of the model group.The damage amount in the acupuncture group was between the blank group and the model group.acupunture can attenuate the effects of D-gal on cognitive function by attenuating apoptotic neurons in the hippocampus.4 Conclusion4.1 To analyze and sort out the electronic literature on the treatment of hearing loss with acupuncture and moxibustion in the past 20 years.The selected acupoints are mainly selected from local acupoints for short distances(Tinggong、Yifeng、Tinghui、Eemen),and mainly acupoints comes from meridian of Hand Shaoyang Sanjiao,Foot Shaoyang Gallbladder,and Hand Taiyang Small Intestine(Zhongzhu、Taixi、Waiguan、Taichong).Great attention to the application of five shu acupoints(Taixi、Xiaxi、Zhongzhu 、 Zulinqi),confluence acupoints(Tinggong 、 Yifeng)and non-specific acupoints(Baihui、Ermen、Tinghui),the selection and compatibility of acupuncture points focous on the methods of "upper and lower acupoints" and "far and near acupoints".4.2 Acupuncture under the guidance of the method of strengthening the spleen and nourishing the kidney is effective in the treatment of ARHL with MCI in terms of hearing and cognitive function.In hearing aspects: Simple changes in living habits and standardization of medication for underlying diseases have little effect on the pure tone hearing threshold,but have a positive impact on the speech recognition rate.the group which Under the guidance of the spleen and kidney method,the effect on hearing is quick increased,which can significantly improve the hearing threshold of patients in 1000 Hz and 2000 Hz frequency,speech recognition rate score and speech recognition rate loss degree,and the change of speech recognition rate is earlier than the change of pure tone hearing threshold.Patients with flat type hearing loss,hearing loss degree and mild speech recognition rate impairment have a better prognosis.In cognitive function: under the guidance of the spleen and kidney method,the Mo CA total score and four aspects of memory and delayed memory,attention,language,visual space and executive ability can be significantly improved.4.3 7 month C57BL/6J mice were injected subcutaneously with D-gal1000mg/Kg·day on the nape of neck for 4 weeks can successfully create a mouse model of ARHL and cognitive dysfunction.D-gal can cause aging of auditory center and hippocampus in 7 month C57BL/6J mice,and may further promote peripheral hearing loss.Acupuncture at Baihui(GV20),Shenshu(BL23)and Zusanli(ST36)under the guidance of the method of strengthening the spleen and nourishing the kidney can delay the progression of presbycusis by reducing the apoptosis of auditory central neurons,and reduce the damage to cognitive function by antagonizing the apoptosis of hippocampus neurons.