The Basic Study Of Different Drugs Intervention On Disc Degeneration And The Measurement Of Inclination Angles Of The L4 And L5 Lamina Ventral Surface And The L4 And L5 Vertebral Body Posterior Surface

Part one Resveratrol Combined with 17β-Estradiol Prevents IL-1β Indu ced Apoptosis in Human Nucleus Pulposus Via The PI3K/AK T/mTOR and PI3K/AKT/GSK-3β PathwayOBJECTIVE:Nucleus pulposus(NP)apoptosis is mainly charged for th e pathological process of Intervertebral disc degeneration(IVDD).Our previo us study Revealed that Resveratrol(RSV)combined with 17β-estradiol(E2)was more effective in cutting down IL-1β induced NP cell apoptosis via P I3K/Akt pathway.And the present study mainly focus on investigating prote ins acting downstream PI3K/Akt in the anti apoptosis process.METHODS:Human nucleus pulposus(NP)cells culture system and IL-1β inducing apoptosis model were constructed in this research.And then th e apoptosis was inhibited by Resveratrol and E2.FACS and CCK-8 assays were respectively used to determine apoptotic incidence and cell via bility of NP cells.Real-time quantitative RT-q PCR was to determine m RNA level of target genes.And Westernblot analysis was performed to detect the chan ges of related protein level.RESULTS:RSV together with E2 attenuate cell apoptosis induced by I L-1β and Recovered cell viability abated by IL1-β,and blockers for mTOR and GSK-3β abate this effect of RSV and E2.RSV combined with E2 inc resed activated p-mTOR and p-GSK-3β obviously,which contribute to the d ownregulation of caspase-3 protein level.whereas,activated p-NF-Kappa B wa s not associated with RSV and E2 anti apoptosis precess.CONCLUSION:This study presented thatcombined use of Resveratrol with E2 effificiently resisted IL-1β induced apoptosis of NP cell,which was mainly through employing PI3K/Akt/mTOR/caspase-3 and PI3K/AKT/GSK-3β/caspase-3 Pathway.Part two Higenamine inhibits IL-1β induced inflammation in human n ucleus pulposus cellsOBJECTIVE:Intervertebral disc degeneration(IDD)is a natural progre ssion of the aging process associated with inflammation.Higenamine,a plan t-based alkaloid,has been identified to possess various pharmacological pro perties,including anti-inflammatory activity.In the present study,we aimed to evaluate the role of higenamine in interleukin(IL-1β)induced inflamm ation in human nucleus pulposus cells(NPCs).METHODS: Human nucleus pulposus(NP)cells culture system and IL-1β inducing inflammatory reaction model were constructed in this researc h.And then the inflammatory reaction was inhibited by Higenamine.MTT as says were respectively used to determine cell viability of NP cells.Real-tim e quantitative RT-q PCR was to determine m RNA level of target genes.And Westernblot analysis was performed to detect the changes of related protei n level.RESULTS:The results showed that higenamine improved cell viability in IL-1β induced NPCs.The IL-1β dependent up-regulation of inflammator y molecules including inducible nitric oxide synthase(i NOS),nitric oxide(NO),prostaglandin E2(PGE2),cyclooxygenase-2(COX-2),tumor necrosis factor alpha(TNF-α),and IL-6 was attenuated by higenamine in NPCs.T he increased productions of matrix metalloproteinases(MMP-3 and MMP-13),as well as a disintegrin and metalloprotease with thrombospondin motif s(ADAMTS-4 and ADAMTS-5)were significantly mitigated by higenamin e treatment.Furthermore,we also found that higenamine suppressed the IL-1β induced activation of NF-kappa B signaling pathway in NPCs.CONCLUSION:the present study proved that higenamine exhibited ant i-inflammatory activity against IL-1β induced inflammation in NPCs via in hibiting NF-kappa B signaling pathway.These results suggested that higena mine might be a therapeutic agent for the treatment of IDD.Part three The measurement of inclination angles of the L4 an d L5 lamina ventral surface and the L4 and L5 vertebral bo dy posterior surfaceOBJECTIVE: To measure the inclination angle of the ventral surface of the lamina(L4,L5)and the posterior surface of the vertebral body(L4,L5),and to explore its clinical significance.METHODS: From January 2019 to March 2020,a 128 row multi-slice spiral CT scanner was used to scan the lumbar spine of 50 patients aged 20-50 years.According to the CT scanning data,the data was processed by the software of mimics 20.3D printer(ANYCUBIC 3D printer)was used for L4 and L5 of each patient to print the 3D model and determine the ventral side of the lamina on the model.Using mimics 20 software,respectively measure the angles of the upper ventral surfaces of L4 bilateral lamina and posterior surfaces of L4 vertebral body(∠HK(L)and ∠HK(R)),the angles of the lower ventral surfaces of L4 bilateral lamina and posterior surfaces of L4 vertebral body(∠JK(L)and ∠JK(R)),angles of the upper ventral surfaces of L5 bilateral lamina and posterior surfaces of L5 vertebral body(∠H’K’(L)and ∠H’K’(R)),the angles of the lower ventral surfaces of L5 bilateral lamina and posterior surfaces of L5 vertebral body(∠J’K’(L)and ∠J’K’(R)).PACS system was used to measure the tilt angle between the ventral surface of lamina and the posterior surface of vertebral body(K,K ’)on L4 and L5 CT scan respectively in axial and sagittal position: the angle of the upper part of the ventral surface of L4 left and right lamina relative to the posterior surface of L4 vertebral body(AL4 and AR4)in axial position,the angle of the lower part of the ventral surface of L4 left and right lamina relative to the posterior surface of L4 vertebral body(AL4’and AR4’)in axial position.the angle of the upper part of the ventral surface of L4 left and right lamina relative to the posterior surface of L4 vertebral body(BL4 and BR4)in sagittal position,the angle of the lower part of the ventral surface of L4 left and right lamina relative to the posterior surface of L4 vertebral body(BL4’and BR4’)in sagittal position.the angle of the upper part of the ventral surface of L5 left and right lamina relative to the posterior surface of L5 vertebral body(AL5 and AR5)in axial position,the angle of the lower part of the ventral surface of L5 left and right lamina relative to the posterior surface of L5 vertebral body(AL5’and AR5’)in axial position.the angle of the upper part of the ventral surface of L5 left and right lamina relative to the posterior surface of L5 vertebral body(BL5 and BR5)in sagittal position,the angle of the lower part of the ventral surface of L5 left and right lamina relative to the posterior surface of L5 vertebral body(BL5’and BR5’)in sagittal position.Kolmogorov Smirnov normality test was used to analyze the distribution of variables.Normal distribution and continuous variables are represented by means±SD,and non normal distribution continuous variables are represented by median(interquartile distance).The independent t-test was used to compare the differences between the two groups of normal distribution variables,and the Mann Whitney U test was used to compare the differences between the two groups of non normal distribution variables.RESULTS: ∠HK(L)(46.37±2.63)°,∠HK(R)(45.28±1.73)°,∠JK(L)(44.83±2.24)°,∠JK(R)(43.92±1.45)°,∠H’K’(L)(36.72±3.21)°,∠H’K’(R)(34.53±4.11)°,∠J’K’(L)(53.35±2.96)°,∠J’K’(R)(52.47±6.86)°.There was no significant difference between∠HK(L)and ∠HK(R)(P>0.05),between∠JK(L)and ∠J K(R)(P>0.05),between∠H’K’(L)and ∠H’K’(R)(P>0.05),between∠J’K’(L)a nd ∠J’K’(R).The difference is statistically significant between∠HK(L)and ∠JK(L)(P<0.05),between∠HK(R)and ∠JK(R)(P<0.05),between∠H’K’(L) and ∠J’K’(R)(P<0.05),between∠H’K’(R)and ∠J’K’(R)(P<0.05).AL4(46.04± 5.25)°,AR4(44.04± 5.29)°,AL4’(50.35± 5.26)°,AR4’(58.78± 4.78)°,AL5(51.87±5.02)°,AR5(51.39±5.92)°,AL5’(43.53±5.62)°,AR5’(49.78±4.92)°,BL4(3.61±8.18)°,BR4(3.85±2.82)°,BL4’(29.64±6.07)°,BR4’(19.64±2.52)°,BL5(11.56±8.83)°,BR5(9.61±4.32)°,BL5’(45.56±8.81)°,BR5’(48.61±4.59)°.There was no significant difference between AL4 and AR4(P>0.05),between AL4’ and AR4’(P>0.05),between AL5 and AR5(P>0.05),between AL5’and AR5(P>0.05).There was significant difference betwee n AL4 and AL5(P<0.05),between AL4’ and AL5’,between AR4 and AR5(P<0.05),between AR4’and AR5’(P<0.05);between BL4 and BR4(P<0.05);be tween BL4’ and BR4’(P<0.05).There was no significant difference between BL5 and BR5(P>0.05),between BL5’ and BR5’(P>0.05);BL5 was signific antly greater than BL4,and the difference was statistically significant(P<0.05),BL5 was significantly greater than BL4’,and the difference was statistica lly significant(P<0.05);BR5 was significantly greater than BR4,and the diffe rence was statistically significant(P<0.05),The difference was statistically sig nificant(P<0.05).CONCLUSION:the anatomic characteristics of L4 and L5 lamina are clear in this study,so as to improve the safety of using circular saw to o pen the window of lamina.The range of inclination angle of L4 and L5 l amina ventral side relative to posterior edge of vertebral body was obtaine d.Compared with L4,the shape of L5 spinal canal and lamina was signifi cantly different and the anatomy of each patient is different.The results of this study must be combined with a customized preoperative surgical plan for clinical patients.